Abstract1
Upcoming SlideShare
Loading in...5
×
 

Abstract1

on

  • 4,308 views

 

Statistics

Views

Total Views
4,308
Views on SlideShare
4,308
Embed Views
0

Actions

Likes
0
Downloads
15
Comments
0

0 Embeds 0

No embeds

Accessibility

Categories

Upload Details

Uploaded via as Adobe PDF

Usage Rights

© All Rights Reserved

Report content

Flagged as inappropriate Flag as inappropriate
Flag as inappropriate

Select your reason for flagging this presentation as inappropriate.

Cancel
  • Full Name Full Name Comment goes here.
    Are you sure you want to
    Your message goes here
    Processing…
Post Comment
Edit your comment

Abstract1 Abstract1 Document Transcript

  • FROM GENOMICS TO PLANT IMPROVEMENT 基因组学与植物改良Proceedings of the 3rd International Conference of Plant Molecular Breeding 第三届植物分子育种国际会议论文摘要 Beijing,September 5-9, 2010 北京,2010 年 9 月 5-9 日
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京ICPMB2010 OrganizationHonorary Presidents Dr. JM Ribaut, Generation Challenge Program, CGIAR Dr. Huqu Zhai, Chinese Academy of Agricultural Sciences Dr. Qifa Zhang, Huazhong Agricultural University Dr. Jiayang Li, Chinese Academy of SciencesPresident Dr. Zhikang Li, Chinese Academy of Agricultural Sciences & International Rice Research InstituteCo-Presidents Dr. Jianmin Wan, Institute of Crop Sciences, Chinese Academy of Agricultural Sciences Dr. Aimin Zhang, Institute of Genetics and Developmental Biology, Chinese Academy of SciencesInternational Organizing CommitteeChair : Zhikang Li, Chinese Academy of Agricultural Sciences & International Rice Research InstituteCo-Chair : JM Ribaut, Generation Challenge Program, CGIARMembers: Aimin Zhang, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences Andrew H. Paterson, University of Georgia, USA Christian Jung, Plant Breeding Institute, Christian-Albrechts-University of Kiel David Mackill, International Rice Research Institute, Philippines Jiayang Li, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences Jinguo Hu, USDA-ARS, USA John Z Yu, USDA-ARS, Crop Germplasm Research, Texas A&M University, USA Lijun Luo, SAGC, Shanghai Academy of Agricultural Sciences Mark J. van Haaren, Keygene N.V. Masahiro Yano, National Institute of Agrobiological Sciences, Japan Graham McLaren, The Generation Challenge Program, CGIAR Henry T. Nguyen, University of Missouri, USA Noel Ellis, John Innes Centre, UK Peter Langridge, Australia National Center for Plant Functional Genomics, Adelaide, Australia Qifa Zhang, Huazhong Agricultural University Roberto Tuberosa, University of Bologna, Italy Swapan Datta, Indian Council of Agricultural Research, India Yunbi Xu, CIMMYT, Mexico Xingwang Deng, Peking University, China; Yale University, USA 2
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京 Zhonghu He, Chinese Academy of Agricultural Sciences & CIMMYT Michael Thomson, International Rice Research Institute, PhilippinesLocal Organizing CommitteeChair : Jianmin Wan, Institute of Crop Sciences, Chinese Academy of Agricultural SciencesCo-Chairs : Aimin Zhang, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences Shuming Wang, Institute of Crop Sciences, Chinese Academy of Agricultural SciencesMembers : Daowen Wang, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences Yuxian Zhu, Peking University, China Zhen Zhu, Institute of Genetics and Developmental Biology, Chinese Academy of SciencesProgram CommitteeChair : Zhikang Li, Chinese Academy of Agricultural Sciences & International Rice Research InstituteCo-Chair: Jinguo Hu, USDA-ARS, USAMembers: Aimin Zhang, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences David Mackill, International Rice Research Institute, Ithaca, New York, USA JM Ribaut, Generation Challenge Program, CGIAR Masahiro Yano, National Institute of Agrobiological Sciences, Japan Mark J. van Haaren, Keygene N.V. Noel Ellis, John Innes Centre, UK Peter Langridge, Australia Center for Plant Functional Genomics Qifa Zhang, Huazhong Agricultural University, China Roberto Tuberosa, University of Bologna, Italy Swapan Datta, Indian Institute of Agricultural Research, India Xingwang Deng, Peking University, China; Yale University, USA Yongbiao Xue, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences 3
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京 ContentsLECTURESPlenary Session I……………………………………………………………….……………..10-13Molecular breeding in developing countries: not a dream anymore. Ribaut JMProgress of rice functional genomics research and the implications in crop genetic improvement. Zhang QFTowards molecular design of super rice. Li JYProgress and challenges in molecular breeding for drought tolerance in crop plants. Nguyen HTPlenary Session II…………………………………………………………….……………..14-18Three genetic systems controlling rice growth and productivity–a reevaluation of the green revolution. Li ZKGenomics-assisted germplasm enhancement and its integration to breeding in rice. Yano MMolecular basis of heterosis in crop plants: From nonadditive gene expression to gene regulatory network.Sun QXTransgenic trait development and deployment circa 2010. Bedbrook JTransgenic crop research in India-current status and perspectives. Datta SPlenary Session III……………………………………………………………….…………..19-23Fostering molecular breeding in developing countries: The GCP approach. Delannay XThe sorghum genome, the diversification of cereals, and the productivity of tropical grasses. Paterson AHPolyploidy and epigenetics: direct application and impact on crop improvement. Chen ZJIdentification of key regulators for flowering time control and their application in breeding of biennial cropspecies. Jung CWhole genome strategies for molecular plant breeding. Xu YBPlenary Session IV…………………………………………………………….……………..24-28Global epigenetic and transcriptional trends among two rice subspecies and their reciprocal hybrid. Deng XWBreeding seeds of innovation. Hervé PM 4
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Breeding by design and innovations in molecular plant breeding. Sorensen AMeeting the challenge of higher nutritional value in seeds: a novel way of increasing methionine content inseeds of the model plant of tobacco. Amir RAssociation mapping for enhancing maize genetic improvement. Yan JBConcurrent session 1: Molecular breeding for abiotic stress tolerances……………….…29-35Mapping QTLs for root morphology in relation to nutrient uptake in wheat. Tong YPThe research progress of drought tolerance and molecular breeding in maize. Wang GYTowards molecular breeding for salt tolerance through modification of root System architecture. Li XMapping and validating QTLs for plant height developmental behaviours in bread wheat. Jing RLDiscovery of genes for drought resistance improvement of rice by systematic genetic and functional genomicapproaches. Xiong LZHeat stress transcriptome analysis and functional characterization of responsive genes in wheat. Ni ZFConcurrent session 2: Gene discovery and function……………………………………….36-42Identification and application of the rice broad-spectrum blast resistance gene Pigm. He ZHMutant resources for functional studies of genes related to fertility in rice. Wu CYGene discovery from common wild rice (Oryza rufipogon Griff). Sun CQDiscovery of brown planthopper resistance gene in rice. He GCMolecular basis of cytoplasmic male sterility in rice. Liu YGToward map-based cloning of a good eating-quality QTL derived from an elite Japanese rice cultivarKoshihikari. Hori KMap-based cloning of QTL genes for flowering time/maturity in soybean. Xia ZJConcurrent session 3: Molecular breeding for biotic stresses…………………….……….43-49From QTLs for fungal disease resistance to marker-assisted selection in durum wheat. Maccaferri MGenomic approaches to plant defense research and crop improvement for insect resistance. Huang YHImprovement of maize resistance to head smut and stalk rot. Xu MLEnhancing broad spectrum resistance to rice diseases. Wang SPMolecular mapping of adult-plant resistance genes to stripe rust and powdery mildew and validation of allelic 5
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京specific markers for Lr34/Yr18/Pm38 in Chinese wheat cultivars. Xia XCInfection character and rice resistance screening of Southern rice black-streaked dwarf virus, a new Fiji virusthreating rice production in Asia. Zhou GHConcurrent session 4: New transgenic technologies, products and markets……………..50-58New transgenic technologies. Broglie RSimultaneously changing several quality traits of Brassica napus by one transgenic event. Liu CLIn situ Pistil Delivery: A High Throughput Method of Brassica Genetic Transformation. Guo XLWheat genetic transformation in China, current status, challenges and future perspectives. Xia LQA new effective selection marker for crop transformation. Xia MEnhancing the lysine in wheat grain by genetic transformation of a lysine rich protein gene Cflr. Ma HXTransgenic strategies for improving drought tolerance traits in chickpea. Bhatnagar-Mathur PIdentification of stress-inducible and tissue-specific promoters in rice. Zhou JLConcurrent session 5: Molecular breeding for cotton, brassica and bio-energy crops…..59-65Progress toward genome sequencing of upland cotton, Gossypium hirsutum. Yu SXMaternal effects and genetic improvement of seed oil content in Brassica napus. Wang HZTowards establishing a molecular breeding platform in cotton: Progress and challenges. Kumpatla SPMolecular breeding of apomixis hickory. Huang JQMining of novel genes for cotton fiber improvement. Yu JZRational design and molecular breeding of sorghum, a dedicated bioenergy crop. Huang YHMolecular breeding for cottonseed quality improvement. Zhu SJMolecular focus in commercial plant breeding. Rossouw JDConcurrent session 6: Maize molecular breeding……………………………………….....66-72QTL fine mapping of leaf angle and leaf orientation value in maize. Chen YHApplication of molecular techniques in maize haploid breeding. Chang MTIdentification of gene marker sets for screening maize lines for resistance to aflatoxin contamination. Luo MMaize disease resistance gene discovery and utilization through association and linkage mapping. Mahuku G 6
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Forward to molecular breeding from genetics in high-oil maize. Li JSGenome-wide association study identifies known as well as novel loci for maize kernel tocopherol content andcomposition. Li QConcurrent session 7: Applied plant genomics: from genomics to field…………………..73-77Molecular breeding in chickpea- still a dream or the reality now! Varshney RKSingle-base resolution DNA methylomes of rice and new regulatory roles of DNA methylation in plant geneexpression. Li XInsertion site-based polymorphism markers open new perspectives for genome saturation and marker-assistedselection in wheat. Paux EIntegrating technologies for genetic improvement of quantitative traits in sorghum. Mace EIrradiation mutant mapping of wild beet translocation lines carrying resistance genes against the beet cystnematode. Capistrano GConcurrent session 8: Rice molecular breeding…………………………………..………..78-85Development of 384-plex SNP marker sets for diversity analysis, mapping, and marker-assisted selection in rice.Thomson MJEpigenetic and genetic control of drought tolerance in rice – a merging story of Larmarkism and Mendelism.Li ZKClustered QTLs for source leaf size and yield traits in rice (Oryza sativa L). Yu SBMolecular breeding approaches for sustainable disease resistance in rice: Current and future strategies.Vera Cruz CMMAS pyramiding of disease and pest resistant genes into drought tolerant hybrid rice. Mei HWDevelopment of single nucleotide polymorphisms (SNPs) detection platforms for genetic analyses andmolecular breeding of rice. Chen HDIdentification of a new blast resistant gene from Dacca6, a useful donor to improve the wide spectrum resistanceof Jin23 against rice blast fungi (Magnaporthe grisea) in Southeast China. Shi BHConcurrent session 9: Wheat molecular breeding…………………………………….……86-93Towards systematic genetic and functional analyses of the complex gliadin gene family in common wheat.Wang DWDevelopment and application of molecular markers for improving processing quality in common wheat. 7
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京He ZHNew insights into the organization, recombination, expression and functional mechanism of low molecularweight glutenin subunit genes at the complex glu-3 loci in bread wheat. Ling HQQTL mapping and marker assisted selection for some quality traits in bread wheat. Gupta PApplication of MAS for resistance to Fusarium head blight in a wheat breeding program Fedak GGenomic distribution of quantitative trait loci (QTL) for yield and yield-related traits in common wheat(Triticum aestivum). Zhang LYGene function and modulation of DREB (dehydration-responsive element binding protein) genes from soybean.Chen MConcurrent session 10: Molecular breeding platform and new technologies…………...94-98The integrated breeding platform: vision and practice. McLaren GOptimization of NGS-based SNP discovery approaches for facilitating molecular breeding in orphan cropspecies. Varshney RISMAB: A data visualization and decision support tool for crop improvement. Shah TBringing genomic data to breeding: what we expect from the IBP to help future breeding. Liang CZDevelopment and optimization of the 50K infinium chip for maize diversity analysis. Ganal MConcurrent session 11: Germplasm and genetic diversity…………………………..…..99-106Core collection-based genomic stocks in wheat. Jia JZHigh-throughput SNP genotyping of a subset of lettuce landraces for genetic diversity assessment. Hu JGThe genetic diversity, structure and classification of rice germplasm in China Li ZCGenetic Diversity Studies on Cool Season Legumes. Zong XXMolecular diversity reveals narrow genetic base of local Ghanaian accessions. Quain MDThe strategy and potential utilization of temperate germplasm for tropical germplasm improvement—a casestudy in maize (Zea mays. L). Wen WWConcurrent session 12: Molecular breeding in legumes and trees crops…………….....107-110Concentration of genetic diversity for gene discovery and broadening genetic base of modern cultivar insoybean. Qiu LJ 8
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Development and application of genomic resources for molecular breeding in groundnut (Arachis hypogaea L).Pandey MThe genomics path from pre-breeding to marker-assisted selection in wheat and barley. Tuberosa RGenomics tools to aid cassava breeding for drought tolerance Rabinowicz PGenetic networks controlling zygomorphic development in legumes Luo DPOSTERS……………………………………………………………………………..….111-231 9
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Plenary Session IMolecular breeding in developing countries: not a dream anymoreRibaut JMGeneration Challenge Programme (GCP), c/o CIMMYT, Int APDO Postal 6-641, 06600 Mexico, DF, Mexico.Email: J.RIBAUT@cgiar.orgMolecular breeding (MB) is definitely an efficient approach, when the necessary minimum humanand operational resources are already in place.This is because MB increases genetic gain per cropcycle, stacksfavourable alleles at target loci and reduces the number of selection cycles. In the lastdecade, the private sector has benefitted immensely from MB, which demonstrates its efficacy. Incontrast, MB adoption is still limited in the public sector, and hardly used in developing countries.Major bottlenecks in these countries include shortage of well-trained personnel, inadequatehigh-throughput capacity, poor phenotyping infrastructure, lack of information systems or adaptedanalysis tools, or simply resource-limited breeding programmes.The emerging virtual platformsaided by the information and communication technology revolution will help to overcome someof these limitations, by providing breeders with better access to genomic resources, advancedlaboratory services, and robust analytical and data management tools. It is unrealistic to projectthat large-scale MB breeding activities will be conducted in the near-term in developing countries.However, the exponential development of genomic resources,the ever-decreasing cost of markertechnologiesand the emergence of platforms for accessing MB tools and support services, plus theincreasing public–private partnerships and needs-driven demand for improved varieties to counterthe global food crisis, are all grounds to predict that MB will have a significant impact on cropbreeding in developing countries. 10
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Plenary Session IProgress of rice functional genomics research and the implications in cropgenetic improvementZhang QFNational Key Laboratory of Crop Genetic Improvement and National Center of Plant Gene Research, HuazhongAgricultural University, Wuhan 430070, ChinaEmail: qifazh@mail.hzau.edu.cnThere has been a large global effort in rice functional genomics research aiming atcharacterization of the full complement of the rice genes. The Chinese program on rice functionalgenomic research is composed of the following components: (1) development of technologicalplatforms, (2) functional genomics of agriculturally important traits, (3) molecular cloning ofimportant genes and, (4) gene discovery by resequencing natural diversity of the rice species. Thetraits targeted for functional genomic studies include yield, grain quality, stress tolerance, diseaseand insect resistances, and nutrient use efficiency. Major progress has been made in a number offronts. Totally 270,000 independent transformants have been generated for the T-DNA insertionmutant library and are now being screened for mutations of important traits. Over 50000 flankingsequences have been isolated, and their analyses identified a number of interesting features ofnonrandom distributions of the T-DNA insertions in the rice genome. A large number of mutantshave now been targeted for gene isolation. For genome-wide expression profiling, data have beengenerated from a large number of tissues covering the whole life cycle of the rice plants grownunder various conditions. Map-based cloning has been applied to isolate genes of agronomicimportance, including dozens of genes for yield, grain quality, fertility restoration, resistances tobiotic and abiotic stresses. Hundreds of accessions of rice germplasm have been resequencedusing new sequencing technologies. The implications of these developments in crop geneticimprovement will be discussed in the presentation. 11
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Plenary Session ITowards Molecular Design of Super RiceLi JY, Wang YHInstitute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, ChinaEmail: jyli@genetics.ac.cnRice (Oryza sativa. L) is one of the most important staple crop, feeding more than half of theworld’s population. To achieve super rice varieties, we focus on the improvement of the grainyield, grain quality, disease and insect resistance. Rice plant architecture, a collection of theimportant agronomic traits that determine grain production, is mainly affected by factorsincluding tillering (tiller number and tiller angle), plant height, and panicle morphology. Toelucidate molecular mechanisms that control rice plant architecture, we have identified severalkey genes that contribute greatly to the plant architecture of rice. Among them, theMONOCULM1 (MOC1) gene was characterized as an essential regulator involved in tiller budinitiation and outgrowth; the DWARF27 (D27) gene acts as a new component involved in thebiosynthesis of strigolactones and controls rice tiller number by regulating the outgrowth of tillerbuds; the LA1 gene plays an important role in determining tiller angle by negatively regulatingpolar auxin transport (PAT); the SHORT PANICLE1 (SP1) gene encodes a transporter thatregulates the panicle size. The quantitative trait locus (QTL) gene, Ideal Plant Architecture 1(IPA1), profoundly affects rice plant architecture and substantially enhances rice grain yield. Ourstudies demonstrate that the application of these genes will facilitate to breed new elite varietiesby modifying tiller number, tiller angle, plant height, panicle morphology and lodging resistance.To improve the rice grain quality, we carried out a systematic examination of geneticdeterminations of rice grain ECQ through a comprehensive association analysis, the results ofwhich were then further have been confirmed by gene transformation. A series of molecularmarkers have been developed for MAS. Our research findings provided a much clearer picture ofhow starch synthesis system regulates grain quality. Also, we engage in cloning insect resistancegenes and developing molecular markers that are linked to quantitative trait loci for rice insectresistance. Our studies will provide a molecular basis for developing super rice varieties in thefuture. 12
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Plenary Session IProgress and challenges in molecular breeding for drought tolerance in cropplantsNguyen HT, Valliyodan B, Manavalan LDivision of Plant Sciences and National Center for Soybean Biotechnology, University of Missouri, Columbia,MO 65211Email: nguyenhenry@missouri.eduProduction of sufficient food for the growing world population during the verge of global climatechanges will be one of the major challenges for the future. This demands the requirement ofdirected adaptation of crop species on an unprecedented magnitude. The global grain demand isexpected to be double by 2050. Much effort is being made by agricultural researchers worldwideto reduce water use by crops to address the challenge which especially affect farmers indrought-prone environments across the developing world. Understanding the concept andcomponents of drought resistance is a key factor for improving drought tolerance of crops.Research to date has shown that improvements in crop drought resistance are from the increasingdehydration avoidance, specifically increasing water availability for plant functions throughchanges such as earlier development, smaller leaves, and deeper roots. In addition, plasticityresponse of root growth under water deficit conditions, and dehydration tolerance traits such as;osmotic adjustment, cell membrane stability, and mobilization of stem carbohydrate reserves incrops also play specific roles in drought resistance mechanisms.Molecular breeding approaches through identification of quantitative trait loci (QTL) andmarker-assisted selection offers an opportunity for significant improvements in the droughttolerance of crops; however the successful application of marker assisted selection to cropbreeding is still in the preliminary stage. Past studies aimed at osmo-protection did not result infield performance for drought tolerance in crops. Recent work on engineering candidate genesincluding transcription factors and cold shock responsive proteins to enhance drought toleranceshowed promising results in field conditions. Transgenic maize plants with a transcription factorshow tolerance to drought based on the responses of a number of stress-related parameters,including; stomatal conductance, leaf temperature, reduced wilting, and maintenance ofphotosynthesis. Another example is engineering farnesylation machinery for plant droughttolerance and yield protection-through stomatal closure, and these transgenic plants showedpromising field performance. Enhanced drought tolerance has also been observed in transgenicplants expressing a cold shock protein under field conditions. Research advances in the area ofintegrated functional genomics will certainly be helpful to improve the molecular breeding andplant transformation approaches to achieve a significant progress in the generation of crop plantswith enhanced drought resistance. 13
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Plenary Session IIThree Genetic systems Controlling Rice Growth and productivity – AReevaluation of the Green RevolutionZhang F1, Xu JL1, Gao YM1, Yu SB2, Fu BY1, Ali J2 and Li ZK1,2 , *1 Institute of Crop Sciences/National Key Facility for Crop Gene Resources and Genetic Improvement, ChineseAcademy of Agricultural Sciences, Beijing 100081, China; 2 IRRI, DAPO Box 7777, Metro Manila, ThePhilippines.*Email: zhkli@yahoo.com.cnThe well-known Green Revolution (GR) since 1960s has more than doubled the productivity of rice, whichresults from loss of function alleles at the GAox-2 locus encoding gibberellin 20-oxidase. Over 95% of thecurrent worldwide rice breeding programs are carried out in the mutant sd1 genetic backgrounds withoutfunctional gibberellin acids (GA). To better understand the effects of sd1 on rice yield and related traits, thephenotypic data of the IR64/Azucena DH population across 11 diverse environments were reanalyzedusing a new molecular-quantitative genetics model. Three genetic systems controlling rice growth andproductivity in rice were revealed, resulting in the discovery of 157 functional genetic units (FGUs)affecting 9 traits related to rice growth, development and productivity. The first one was the GA-mediatedpathways controlled by SD1 and its 43 downstream FGUs for increased plant height (PH), increasedbiomass, reduced spikelet fertility (SF), delayed heading (HD), reduced harvest index (HI), reduced paniclenumber (PN), increased grain weight (GW) and reduced yield. Their effects gain yield (GY) and spikeletnumber per panicle (SN) varied depending on the environments. Of these downstream FGUs, 3 PH QTLs(QPh2b, QPh3b and QPh4a) had effects highly correlated with the mean PH values of the SD1subpopulation, suggesting their positive responses to the overall soil fertility levels of the test environments.Together, the GA-mediated pathways explained 38.6%, ranging from 16.0% for SF to 54.8% for PH. Thesecond system was the GA-repressed pathways that were expressed only in the mutant (sd1) background,which comprised of 39 FGUs for PH, SF, biomass, HD, SN, PN, HI, GW, and yield. The effect directionsof most these pathways could not be determined based on available QTL information. The GA-repressedpathways collectively explained 32.3% of the total genotypic variation of the 9 traits in the DH population,ranging from 14.7% for PN to 59.3% for SN. The third one was the GA independent pathways controlledby 75 FGUs that affected all measured traits. Together, the GA-independent pathways explained 29.2% ofthe total genotypic variation of the 9 traits in the DH population, ranging from 6.0% for PH to 55.8% forPN. Because the overall effects of the GR are reflected by the differences between the GA-mediated andGA-repressed pathways, detailed Comparison between them indicated that the former had larger effects onPH, HD, PN, HI and GY, whereas the latter influenced more SN and SF. Based on these results, theadvantages and potential consequences of the GR gene, sd1, were discussed in the context of the globalrice improvement and its challenges. Alternative breeding strategies for developing “Green Super Rice”cultivars that have high yield potential with less input are proposed based on our discoveries. 14
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Plenary Session IIGenomics-assisted germplasm enhancement and its integration to breeding inriceYano M*, Hori K, Uga Y, Fukuoka S, Ebana K, Yonemaru J and Yamamoto TQTL Genomics Research Center, National Institute of Agrobiological Sciences, Tsukuba, Japan.*E-mail: myano@nias.affrc.go.jpProgresses on recent genomics in rice have provided a new tools and opportunities to enhanceactivity in crop improvement. Elucidation of the association between nucleotide and phenotypicchanges is inevitable to this end and has been a big challenge in molecular genetics and breedingof rice. Toward this goal, we have been involved in the genetic dissection of natural phenotypicvariations in rice and have identified several genes involved in complex traits, including headingdate, shattering habit, pre-harvest sprouting, root morphology, disease resistance, seed size andeating quality. To enhance the power of genetic dissection of complex phenotypes, we aredeveloping several mapping populations, such as recombinant inbred lines and chromosomesegment substitution lines, which will allow us to extract the useful alleles from natural variants.Recently, QTL for durable resistance to rice blast has been cloned from Japanese upland rice. Thisfinding has opened new opportunity to introduction of the unique blast resistance gene without alinkage drag of low eating quality. We have also detected a major QTL for deeper rooting onchromosome 9. This finding has open new opportunity to enhance drought avoidance in rice. Tofacilitate allele mining using novel plant materials, we have also embarked on the genome-widediscovery of single nucleotide polymorphisms (SNPs). In particular, to overcome a shortage ofSNPs among temperate japonica cultivars, we have attempted whole-genome sequencing ofseveral Japanese cultivars using next-generation sequencing approaches. This SNP discovery hasled to the development of an array-based SNP genotyping system in Japanese rice cultivars.Large-scale genotyping of these SNPs has made it possible to visualize pedigree haplotypes ofparticular chromosome segments in the Japanese landraces and modern cultivars. These efforts ingenomics have opened up new opportunities to accelerate not only the genetic dissection ofcomplex traits, but also integration of genomics to breeding in rice. 15
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Plenary Session IIMolecular basis of heterosis in crop plants: From nonadditive gene expressionto gene regulatory networkSun QX, Ni ZF, Yao YY, Peng HR, Du JKChina Agricultural University, Beijing 100183, China.*Email: qxsun@cau.edu.cnWhole genome expression analysis in hybrid and its parental inbreds provides a platform to identifynonadditively expressed genes in hybrids, which have given some insights into the understanding ofmechanisms of heterosis. In this study, two wheat (Triticum aesticum L.) hybrid F1 derived from same femaleparent but displaying contrasting heterosis in primary root are used for expression analysis by using wheatgenome array. The expression polymorphism analysis between the parental inbreds indicates that up to 4%genes display expression difference, but more than 3 times more present-absent genes between the two parentalinbreds are detected in highly heterotic Hybrid A than in nonheterotic Hybrid B. Differential expression (DE)analysis in hybrids and their parental inbreds identify 1019 (4.94%) and 698 (3.23%) DE genes in Hybrid A andB, respectively. It is interesting to note that heterotic Hybrid A tends to have more DE genes of dominance andpartial dominance expression modes than nonheterotic Hybrid B which, however, tends to have more DE genesof negative partial dominance expression mode. By adopting the “Wooden Barrel Principle”, we propose thataccumulation of dominance and partial dominance expression in wheat hybrid could be a major determinant ofroot heterosis. We also find that a substantial number of stress-related genes as well as retrotransposon-like andtransposon-like genes are also included in the DE genes. We propose that as compared to the interspecifichybridization which can be a source of genomic shock as described by Barbara McClintock, hybrids derivedfrom less distantly-related two inbreds can be a source of “mild genomic shock” or “intrinsic stress” in thehybrid genome, which, in turn, could cause expression changes of genes, especially stress-related genes andretrotransposon. Heterosis in internode elongation and plant height are commonly observed in hybrid plants, andhigher GAs contents were found to be correlated with the heterosis in plant height. By using the uppermostinternode tissues of wheat, we examined expression patterns of genes participating in both GA biosynthesis andGA response pathways between a hybrid and its parental inbreds. Our results indicated that among the 18 genesanalyzed, genes encoding enzymes that promote synthesis of bioactive GAs, and genes that act as positivecomponents in the GA response pathways were up-regulated in hybrid, whereas genes encoding enzymes thatdeactivate bioactive GAs, and genes that act as negative components of GA response pathways weredown-regulated in hybrid. Moreover, the putative wheat GA receptor gene TaGID1, and two GA responsivegenes participating in internode elongation, GIP and XET, were also up-regulated in hybrid. A model for GAand heterosis in wheat plant height was proposed. This model is also validated by using 16 wheat hybrids withdifferent level of heterosis in plant height. Our results provided molecular evidences not only for the higher GAlevels and more active GA biosynthesis in hybrid, but also for the heterosis in plant height of wheat and possiblyother cereal crops. Moreover, overexpression of 6 differentially expressed genes suggested that up-regulatedgenes in hybrids could enhance the trait performance but the down-regulated genes in hybrids can have negativeeffects on the trait performance. 16
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Plenary Session IITransgenic Trait Development and Deployment Circa 2010Bedbrook JVice President, DuPont Agricultural BiotechnologyTransgenic traits providing weed and insect pest control solutions, first introduced in the mid1990’s have been rapidly adopted globally in corn, soybean and cotton. Next generationtransgenic traits providing new functionalities, including; grain quality attributes, abiotic stresstolerance, disease resistance and seed production systems are close to commercialization. In thispaper I describe DuPont’s approaches to genetic based gene discovery, event selection, traitdevelopment and commercial deployment for these next generation traits. 17
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Plenary Session IITransgenic Crop Research in India-Current status and perspectivesDatta SKCrop Science Division, ICAR, Krishi Bhavan, New Delhi-110114, IndiaEmail: swpndatta@yahoo.comFunctional genomics provides powerful tool for the identification of desirable genes and theirintroduction into crops for the trait improvement. The ability to introduce beneficial genes underthe control of specific promoters through transgenic approaches is the path towards targeted cropimprovement. Development and commercialization of transgenic crops expressing a wide range ofagronomic traits during mid-nineties has virtually revolutionized the face of global agriculture.Safety of transgenic crops, especially GM food crops is a major concern. To address all the issuesrelated to biosafety, environmental safety, risk assessment, biodiversity and socio-economicimpact the GM crops, Government of India has entrusted the task to the Ministry of Science andTechnology to develop one window regulatory mechanism to approve and release the GM in thefield through NBRA (National Biotechnology Regulatory Authority). The environmental releaseof transgenic cotton with insect-pest resistance in 2002 is a landmark in Indian agriculture. It hasplaced India at the forefront of global cotton production and trade. At the global level, cultivationof transgenic crops in the past twelve years has conferred significant social, economic andenvironmental benefits to mankind. Such a sea change in the production of major food crops is theneed of the hour. Bt cotton, which confers resistance to important insect pests of cotton, was firstadopted in India as hybrids in 2002. The number of events, as well as the number of Bt cottonhybrids and companies marketing approved hybrids increased from one event and 20 hybrids in2005 by more than three-fold in 2009 to six events and 282 hybrids. India currently produces >30million bales of cotton per year and occupies # 2 position in terms of global cotton productionand now #1 in Bt cotton areas. Other Crops such as Bt rice, Bt brinjal, transgenic tomato,Sorghum, Brassica, Groundnut etc are at the different stages of development. 18
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Plenary Session IIIFostering Molecular Breeding in Developing Countries -the GCP approachXavier DelannayGeneration Challenge Program, CGIAREmail:x.delannay@cgiar.orgAn important focus of the Generation Challenge Programme (GCP) since its inception has been topromote an increased use of molecular marker technologies in developing country breedingprogrammes. This started with the implementation in applied breeding programmes ofmarker-assisted selection for new important traits that had been mapped with funding assistancefrom the GCP. More recently, the GCP has focused on the implementation of new integratedbreeding programmes in developing country crops through the use of molecular breedingtechnologies such as marker-assisted recurrent selection (MARS). The use of MARS should helpaccelerate the improvement of crops growing under suboptimal conditions of Africa and Asia,which is also a focus of the GCP. This development will be greatly facilitated by the IntegratedBreeding Platform that is concurrently being developed by the GCP. Examples will be shown ofpractical applications of molecular breeding being used or being put in place in developingcountries for crops such as rice, cassava, sorghum, cowpea and chickpea. 19
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Plenary Session IIIThe sorghum genome, the diversification of cereals, and the productivity oftropical grassesPaterson AHPlant Genome Mapping Laboratory, University of Georgia 111 Riverbend Road, Rm 228, Athens, GA 30602Email: paterson@plantbio.uga.eduSorghum, an African grass related to sugarcane and maize, is grown for food, feed, fiber, and fuel,is representative of tropical grasses that are among the most efficient biomass accumulators thanksto ‘C4’ photosynthesis. An initial analysis of the sorghum genome placed ~98% of genes in theirchromosomal context using whole genome shotgun sequence validated by genetic, physical, andsynteny information. Genetic recombination is largely confined to about one-third of the sorghumgenome with gene order and density similar to those of rice. Retrotransposon accumulation inrecombinationally-recalcitrant heterochromatin explains the ~75% larger genome size of sorghumthan rice. While gene and repetitive DNA distributions have been preserved sincepaleopolyploidization ~70 million years ago, most duplicated gene sets lost one member beforesorghum/rice divergence. Concerted evolution makes one duplicated chromosomal segmentappear only a few million years old. About 24% of genes are grass-specific and 7% aresorghum-specific. Recent gene and miRNA duplications may contribute to sorghum’s droughttolerance. The sorghum sequence offers new means to improve sorghum itself and new or existingbiofuel crops, and to try to control weedy and invasive plants. 20
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Plenary Session IIIPolyploidy and epigenetics: direct application and impact on cropimprovementChen ZJInstitute for Cellular and Molecular Biology, The University of Texas at Austin, Texas 78712, USA.Email: zjchen@mail.utexas.eduPolyploidy, or whole-genome duplication (WGD), is common in some animals and many plants,including important crops such as wheat, cotton, canola, sugar cane, and switchgrass. Thecommon occurrence of polyploidy suggests an evolutionary advantage of having multiple sets ofgenetic material for adaptive evolution and crop domestication. However, increased gene andgenome dosages in autopolyploids (duplications within species) and allopolyploids (combinationof two or more divergent genomes among species) often cause genome instabilities, chromosomeimbalances, regulatory incompatibilities, and reproductive failures. Therefore, new allopolyploidsmust establish a compatible relationship between alien cytoplasm and nuclei and between twodivergent genomes, leading to rapid changes in genome structure, gene expression, anddevelopmental traits such as fertility, inbreeding, apomixis, flowering time, and hybrid vigor.Although the underlying mechanisms for these changes are poorly understood, some themes areemerging. There is compelling evidence for epigenetic changes during early stages of polyploidformation. Using Arabidopsis allopolyploids and hybrids as model systems, we found thatchanges in cis- and trans-acting effects, chromatin modifications, RNA-mediated pathways, andregulatory networks modulate differential expression of homoeologous genes and phenotypicvariation such as flowering time. We have shown that nonadditive gene expression, small RNAs,and epigenetic regulation of circadian-mediated metabolic pathways, play central roles in growthvigor in hybrids and allopolyploids. Understanding epigenetic mechanisms for polyploidy andhybrid vigor will facilitate the use and exploitation of the increased biomass and yield in hybridsand allopolyploids for food, feed, and fuels. 21
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Plenary Session IIIIdentification of key regulators for flowering time control and their applicationin breeding of biennial crop speciesJung C*, Wafa SAE, Büttner B, Schulze-Buxloh G, Müller APlant Breeding Institute, Christian-Albrechts-University of Kiel*Email: c.jung@plantbreeding.uni-kiel.deFloral transition is a major developmental switch that is tightly controlled by regulatory pathwaysthat integrate endogenous and environmental cues to ensure flowering under favourableconditions. Sugar beet (Beta vulgaris) is a biennial crop which bolts and flowers after a period ofcold temperatures over winter, however annual types without vernalization requirement exist. Wehave identified >30 flowering time regulators from the beet genome by different approaches. Theexistence of an FLC-like gene in beet suggests similar regulatory pathways as in Arabidopsis. In acomplementary approach additional components of the floral transition gene network in sugarbeet are being identified by homology to genes from model species and genome-wide transcriptprofiling. We found a number of ESTs with homology to Arabidopsis genes. Using RACE andBAC cloning we identified full length cDNA and genomic sequences. We functionallycharacterized these sequences by expression analysis and transformation into Arabidopsis. Wefound evidence for the existence of autonomous and vernalization pathways in beet similar toArabidopsis, however substantial differences between both species exist. Annuality is controlledby the bolting locus B. We have identified by map based cloning sequences from the B locus withhomology to floral transition genes from other species that suggest that they mediate bolting timecontrol in response to environmental cues. Another QTL for early bolting was mapped withmolecular markers demonstrating for the first time that at least two loci cause early bolting inbeets. A beet TILLING platform now also enables the identification of mutants and functionalcharacterization of candidate genes. New beet prototypes with altered vernalization requirementhave been produced either by EMS mutagenesis or transformation. These mutants in combinationwith transgenic beets with altered bolting behaviors are needed for the breeding of winter beetswhich are sown before winter. Apart from winter hardiness these beets must be completely boltingresistant to prevent bolting after winter. Different approaches to establish fully bolting resistantbeet prototypes are presented. 22
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Plenary Session IIIWhole Genome Strategies for Molecular Plant BreedingXu YB1*, Lu YL2 and Gao SB21 Institute of Crop Science and CIMMYT, Chinese Academy of Agricultural Sciences, National Key Facilitiesfor Crop Genetic Resources and Improvement, 12 Zhongguancun South St., Beijing 100081, China. 2MaizeResearch Institute, Sichuan Agricultural University, Ya’an, Sichuan 625014, China.*Email: y.xu@cgiar.orgMolecular breeding for complex traits in plants needs to understand and manipulate many factorsinfluencing plant growth and development including genotypes, environments and theirinteraction. Molecular breeding procedures can be facilitated and revolutionized through wholegenome strategies, which are featured by utilizing full genome sequence and genome-widemolecular markers to address all genomic and environmental factors through a representative orcomplete set of genetics and breeding germplasm. The strategies should be developed forunderstanding specific genomic region, genes, haplotypes, linkage disequilibrium block or allelesand their contribution to specific phenotypes and breeding products. Genotyping-by-sequencingand genomewide selection are two important components of the strategies. These strategies needto be integrated with precision phenotyping and powerful population management systems.Examples of such integrated systems include joint linkage-linkage-disequilibrium mapping formarker development and gene discovery, breeding-to-genetics approaches by using existinggenetic and breeding materials, and simultaneous genomewide improvement for multiple traits.As components of whole genome strategies, molecular breeding platforms and methodologiesshould be backed up with strong supporting systems such as breeding informatics and decisionsupport tools. Some basic strategies will be discussed using maize as an example, including (1)seed DNA-based genotyping for simplifying marker-assisted selection, reducing breeding cost andincreasing scale and efficiency, (2) selective genotyping and phenotyping for capturing mostimportant contributing factors with optimized breeding design, (3) flexible genotyping systemsrefined for different selection methods including marker assisted selection, marker assistedrecurrent selection and genomic selection, and (4) sequence-based strategies for markerdevelopment, allele mining, gene discovery and molecular breeding. 23
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Plenary Session IVGlobal Epigenetic and Transcriptional Trends among Two Rice Subspecies andTheir Reciprocal HybridsHe GM, Zhu XP, Elling AA, Chen LB, Chen RS and Deng XW*Peking-Yale Joint Center of Plant Molecular Genetics and Agrobiotechnology, College of Life Sciences, PekingUniversity, Beijing 100871, China. Department of Molecular, Cellular and Developmental Biology, YaleUniversity, New Haven, CT 06520, USA. Institute of Biophysics, Chinese Academy of Sciences, 15 DatunRoad, Beijing 100101, China*Email: xingwang.deng@yale.eduThe behavior of transcriptomes and epigenomes in hybrids of heterotic parents is of fundamentalinterest. Here we report highly integrated maps of the epigenome, mRNA and small RNAtranscriptomes of two rice subspecies and their reciprocal hybrids. We found that gene activitywas correlated with DNA methylation and both active and repressive histone modifications intranscribed regions. Differential epigenetic modifications correlated with changes in transcriptlevels among hybrids and parental lines. Distinct patterns in gene expression and epigeneticmodifications in reciprocal hybrids were observed. Through analyses of single nucleotidepolymorphisms from our sequence data, we observed a high correlation of allelic bias ofepigenetic modifications or gene expression in reciprocal hybrids with their differences in theparental lines. The abundance of distinct small RNA size classes differed between the parents andmore small RNAs were down-regulated than up-regulated in the reciprocal hybrids. Together, ourdata reveal a comprehensive overview of transcriptional and epigenetic trends in heterotic ricecrosses, and provides a very useful resource for the rice community. 24
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Plenary Session IVBreeding Seeds of InnovationHervé PMBayer Cropscience, Bioscience NV BelgiumAt Bayer Cropscience, we help farmers worldwide meet the ever-increasing demand foraffordable and high quality food, feed, fiber and energy crops. We aim at providing sustainablecrop solutions from seed to harvest, with outstanding seeds and modern crop protection products.Major technology platforms based on the complementary of modern breeding methods and plantbiotechnology are used to develop new seeds and innovative traits solutions. An update of ourSeeds & Traits Pipeline and key examples of successful molecular breeding solutions for our corecrops will be presented. 25
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Plenary Session IVBreeding by design and innovation in molecular plant breedingSørensen AP, van Schriek M, Hofstede R, Guerra J, Prins M and Buntjer JBKeygene N.V., Agro Business Park 90, P.O. Box 216, 6700 AE Wageningen, the NetherlandsSince the concept of Breeding by Design (BBD) was launched by KeyGene in 2003, DNAtechnologies have developed with a dramatic acceleration; especially high-throughput sequencingtechnologies are revolutionizing the DNA research arena. The possibilities for genetic research toelucidate the molecular mechanism of phenotypic expression have increased significantly. As aconsequence, implementing BBD or BBD like approaches for trait and variety improvementprograms is ongoing.We will discuss here a selection of current genomic tools and applications. Whole genomesequence scaffolds and whole genome BAC based physical maps of commercial crop species arebeing developed, following the examples of model plant organisms. The discovery of totalgermplasm variation at the genotypic level and at the gene haplotype level is practically feasiblefor many crop species. Phenotypic evaluation of germplasm variability is performed with highprecision digital imaging systems and supported by statistical tools for evaluation ofreproducibility, heritability and interrelatedness of phenotypic scores.The current challenge for plant geneticists clearly lies in the ability to integrate and aggregate thedifferent and large data sources, in order to make firm and robust associations between thephenotypic variability and the genotypic variability, after which these can immediately beexploited by modern plant breeders. Furthermore novel technologies for generation of mutantalleles of interesting plant genes are in development and will increase the genetic variability ofgermplasm available for variety improvement programs. We will present some of the approachestaken by Keygene to assist plant breeders in these novel opportunities. 26
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Plenary Session IVMeeting the challenge of higher nutritional value in seeds: a novel way ofincreasing methionine content in seeds of the model plant of tobaccoGodo I, Matityahu I, Hacham Y, Amir RLaboratory of Plant Science, Migal Galilee Technology Center, P.O. Box 831, Kiryat Shmona 12100, IsraelThe sulfur-containing amino acid, methionine, is an essential amino acid whose level limits the nutritionalvalue of crop plants. Yet, aside from its nutritional importance, methionine is also a fundamentalmetabolite in plant cells because it indirectly regulates a variety of cellular processes as the precursor ofS-adenosyl methionine (SAM). This study describes the first modification of methionine biosynthesis inseeds using the model plant, tobacco (Nicotiana tabacum). Overexpression of the unregulated form ofcystathionine gamma synthase (AtD-CGS), the first unique enzyme of methionine biosynthesis pathwayfrom Arabidopsis in tobacco plants, led to an over 10-fold increase in methionine content. However, inthese transgenic plants, the methionine level inside their seeds increased only by 15% compared towild-type seeds. Similar results were obtained when AtD-CGS was seed-specific expressed in tobaccoplants. This suggests that the CGS expression level does not limit methionine synthesis in tobacco seeds.To further study the factors regulating methionine synthesis in seeds, the receptacle of developing podswere fed with homoserine, the substrate of CGS. Seeds from these pods demonstrated three-fold higherlevels of methionine, suggesting that homoserine content limits methionine synthesis. To further test thisassumption, we next crossed between plants seed specific expressing AtD-CGS with those seed-specificexpressing the feedback-insensitive bacterial aspartate kinase (bAK), which evidence suggests their seedshave a higher homoserine content. Seeds obtained from the progenies of this cross showed a three-foldhigher level of methionine compared to wild-type seeds. In addition, the level of threonine, an importantessential amino acid that limits the nutritional quality of cereals, accumulated significantly in these seeds.Our next goal was to reveal if the developing transgenic seeds are tolerant to metabolic perturbations thatoccur with changes in methionine and threonine levels. To this end, we performed metabolic profiling towild-type and transgenic seeds expressing AtD-CGS, bAK and AtD-CGS/bAK using GC-MS. A principalcomponent analysis of about 150 metabolites from each transgenic line shows that these lines differsignificantly from one another. Of these metabolites, only 12 compounds significantly changed andcontributed to this diversity. These include the main amino acids, glutamine and asparagine, and severalsugars, trehalose, galactose, glycerol and melbiose. A further study should be performed to reveal therelationships between these metabolites and methionine metabolism. In general, this study demonstrates anovel way of increasing methionine content in seeds, which consequently contributes to enhancing theirnutritional value. 27
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Plenary Session IVAssociation mapping for enhancing maize genetic improvementYan JB 1,2 *, Li JS 21 International Maize and Wheat Improvement Center, (CIMMYT), Apartado Postal 6-640, 06600 Mexico, DF,Mexico China2 National Maize Improvement Center of China, CAU, Beijing 100193, ChinaE-mail: j.yan@cgiar.orgAssociation mapping through linkage disequilibrium (LD) analysis is a powerful tool for thedissection of complex agronomic traits and for the identification of alleles that can contribute tothe enhancement of a target trait. With the developments of high throughput genotypingtechniques and advanced statistical approaches as well as the assembling and characterization ofmultiple association mapping panels, maize has become the model crop for association analysis.In this talk, we summarize the progress in maize association mapping and the impacts of geneticdiversity, rate of LD decay, population size and population structure. We also report the use ofcandidate genes and gene-based markers in maize association mapping studies which hasgenerated particularly promising results. In addition, we examine recent developments ingenome-wide genotyping techniques which promise to improve the power of association mappingand significantly refine our understanding of the genetic architecture of complex quantitative traits.Already these seem to be suggesting that the structure of agronomic traits in maize has more incommon with important traits in humans and animals than it does with similar traits inArabidopsis and rice. The new challenges and opportunities associated with genome-wideanalysis studies will be discussed. 28
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 1: Molecular breeding for abiotic stress tolerancesMapping QTLs for root morphology in relation to nutrient uptake in wheatHe X, Li JJ, Ren YZ, Zhao XQ, Li B, Li ZS and Tong YP*State Key Laboratory for Plant Cell and Chromosome Engineering, Institute of Genetics and DevelopmentalSciences, Chinese Academy of Sciences, Beijing 100101.*Email: yptong@genetics.ac.cnNitrogen (N) and phosphorus (P) fertilizers are required to maximize crop yields in manyagricultural systems. However, the recovery rates of fertilizer N and P were low. To increase Nand P recovery rates, systematic approaches are required, including optimizing managementpractices and breeding crops with improved N and P use efficiency.Previous studies have shown that vigorous early root growth is a major factor influencing N and Puptake in wheat. However, roots, the ‘unseen half’ of wheat plants, are difficult to be selecteddirectly by wheat breeders. Therefore, identifying QTLs/genes regulating root traits can helpwheat breeders to develop wheat varieties with ideal root system for efficient use of nutrientsthrough MAS approach.A RIL population of derived from two Chinese wheat varieties Xiaoyan 54 and Jing 411 was usedto map QTLs for root traits in relation to N and P uptake. A hydroponic culture and a soil columnexperiment were carried to phenotype the RIL population at seedling stage. For the hydroponicculture, the maximal root length (MRL), root dry weight (RDW), shoot dry weight (SDW), N(NUP) and P (PUP) uptake of this RIL population were investigated under sufficient nutrientsupply, low N and low P conditions. Phenotype variation explained by individual QTL variedfrom 4.6% to 32.7%. For the soil column experiment, root distribution in the soil profiles, SDW,NUP and PUP were investigated under sufficient nutrient condition. Phenotype variationexplained by individual QTL varied from 5.2% to 22.5%. To develop MAS for breeding wheatroot traits, we analyzed the effects of pyramiding multi-QTLs on RDW, as well as SDW, NUP andPUP investigated in these tow experiments. The results showed that pyramiding the three QTLslinked with Xgwm157-2D, Xgwm533.2-3B and Xbarc90-4B, respectively, significantly increasedRDW, SDW, NUP and PUP under different N and P supply levels in the hydroponic culture. TheRILs harboring the positive alleles at these three loci had, averagely, 33%-69% higher SDW,RDW, NUP and PUP than those with the negative alleles under different N and P conditions. Inthe soil column experiment, pyramiding the three QTLs linked with Xgwm157-2D,Xgwm533.2-3B and Xbarc70.1-4A, respectively, significantly increased SDW, RDW, NUP andPUP. The RILs harboring all the three positive alleles had, averagely, 30% higher SDW, 25%higher RDW in the 0-30 cm soil layer, 48% higher RDW in the 30-60 cm soil layer, 43% higherRDW in the 60-90 cm soil layer, 31% higher total RDW, 31% higher NUP and 30% higher PUPthan those with the negative alleles. 29
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 1: Molecular breeding for abiotic stress tolerancesThe research progress of drought tolerance and molecular breeding in maize *Zheng J, Fu JJ, Liu YJ, Jian M, Wang GYInstitute of Crop Sciences and National Center for Plant Gene Research, Chinese Academy of AgriculturalSciences, Beijing 100081, China.* Email: gywang@caas.net.cnDrought stress greatly affects maize growth and its yield potential. In order to understand themolecular basis in response to drought stress, and further to improve the drought tolerance inmaize, transcriptome analysis, QTL mapping and transgenic approaches were performed in ourlab. Genome-wide gene expression profiling was analyzed between the drought-tolerant lineHan21 and drought-sensitive line Ye478. Our data identified a common set of ~2,600 regulatedgenes under drought stress between the two lines, and showed that the drought tolerant line hasfewer genes with altered expression. The potential components of the abscisic acid signalingpathway were significantly identified from the common set of genes. A total of 827 genes withsignificantly differential expression between the two lines under drought stress were identified. AF2 population of Han21×Ye478 was used to construct the genetic linkage map and QTL mapping.Drought tolerant NILs (near-isogenic lines) were also screened out from the backcross populationof Han21×Ye478 under severe drought stress conditions. Additionally, the transgenic maize thatoverexpressed HDG11, which encodes a homeodomain-START transcription factor, hadincreased the drought tolerance with improved maize root system and reduced stomatal density. 30
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 1: Molecular breeding for abiotic stress tolerancesTowards molecular breeding for salt tolerance through modification of rootSystem architectureZhao YK, Wang T, Wang ZJ and Li X* Plant Cell & Chromosome Engineering, Center of Agricultural Resources Research, Institute of Genetics andDevelopmental Biology, 286 Huaizhong Road, Shijiazhuang, Hebei, P.R. China.*Email: xli@genetics.ac.cnSalinity is a major constraint to crop growth and production. Root system architecture has beenconsidered as one of most important traits of crops in response to various abiotic stresses.Research on root traits is a major breeding objective in genetic improvement in nutrient useefficiency and drought tolerance. Some quantitative trait loci (QTLs) and genes conferringsuperior root system architecture have been identified. However, the role of developmentalplasticity of root system architecture under salt stress is largely unknown, and the genes and QTLsmediating this trait remains to be identified. To investigate the response of plant root system tosalt stress, we have conducted a systematic study using Arabidopsis plants. We found that the rootsystem architecture is highly sensitive to salt stress. The SOS (Salt Overly Sensitive) genes areessential for root plastic development in response to salt stress. Loss of function in the SOS genesare hypersensitive to salt, particularly the mutant plants exhibited developmental failure in lateralroot initiation and emergence. In contrast, the transgenic plants overexpressing the SOS genesshowed enhanced tolerance to salt stress and developed more root mass. Further, we haveidentified the STS1 (Sensitive To Salt1) gene as an upstream regulator in the root traits mediatedby the SOS signaling pathway in response to salt stress. STS1 gene encodes a WD40 repeatprotein and is induced by salt stress. Interestingly, STS1 interacts with ABI2, a key regulator ofABA signaling pathway, suggesting that ABA may play an important role in the root trait. TheSOS and STS1 genes are found to be conserved in Arabidopsis and winter wheat. The functions ofthe genes and the SOS and ABA signaling in developmental plasticity of root system architecturein various winter wheat with different salt tolerance are under investigation. These results willfurther our understanding of the genetics of salt tolerance in crops and to provide novel insightsinto improvement of their performance under salt stress conditions. 31
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 1: Molecular breeding for abiotic stress tolerancesMapping and validating QTLs for plant height developmental behaviours inbread wheatWu XS, Wang ZH, Zhang JN, Wei TM, Shi W, Zhang B, Jing RL*The National Key Facility for Crop Gene Resources and Genetic Improvement; Institute of Crop Science,Chinese Academy of Agricultural Sciences, Beijing 100081, China.* Email: jingrl@caas.net.cnPlant height (PH), a crucial trait related to yield potential in crop plants, is known to be typicallyquantitatively inherited. However, its full expression can be inhibited by a limited water supply.As a trait easily measured, plant height is also a suitable model trait for exploring droughttolerance from jointing stage to flowering time in wheat (Triticum aestivum L.). In this study, wemapped and validated QTLs for plant height developmental behaviours in wheat by a doubledhaploid (DH) population, a recombinant inbred line (RIL) population, a collection of accessionsand backcross lines. The genetic basis of the developmental behaviour of PH was assessed in a150-line doubled haploid population (Hanxuan 10 × Lumai 14) grown in 10 environments (year ×site × water regime combinations) by unconditional and conditional quantitative trait locus (QTL)analyses in a mixed linear model. QTLs with additive and epistatic effects that expressedselectively during ontogeny were identified. Total of seven genomic regions covering PH QTLclusters on different chromosomes identified from the DH population were used as the candidateQTLs and extensively analyzed in a RIL population derived from the same cross as the DH. Fiveadditive QTLs and eight pairs of epistatic QTLs significantly affecting plant height developmentwere detected by unconditional QTL mapping method. Six additive QTLs and four pairs ofepistatic QTLs were validated using conditional mapping approach. Among them, three additiveQTLs and three pairs of epistatic QTLs were common QTLs detected by both methods. ThreeQTLs were expressed under both drought and well-water conditions. Total of 270 historical winterwheat accessions planted in northern China were genotyped using 60 PH candidate markers on sixchromosomes. A list of association was identified in the regions of gene Rht, indicating aconsistency of association analysis with linkage mapping. A total of 68 backcross lines of BC3F3-4were used to validate the QTLs detected in the genetic populations and natural collection. Theresults showed that some lines pyramiding multi-allele with effect of increasing or decreasingplant height exhibited superiority over the opposite lines. This case, mapping and validating QTLsfor plant height developmental behaviours in wheat indicates the possibility of molecular breedingfor plant complex quantitative traits. 32
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 1: Molecular breeding for abiotic stress tolerancesDiscovery of genes for drought resistance improvement of rice by systematicgenetic and functional genomic approachesXiong LZNational Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan 430070,ChinaE-mail: lizhongx@mail.hzau.edu.cnDrought resistance is a very complex trait with distinct molecular and physiological mechanismsin different plant species. Irrigated rice has been domesticated in full irrigation ecosystem and it isextreme sensitive to drought. With a long-term goal of improving drought resistance in irrigatedrice, we have adopted a strategy by integrating the approaches including germplasm exploitation,genetic and functional genomics approaches to identify loci/genes effective for drought resistanceimprovement of rice. In this paper, we described the approaches and the major progresses made todiscover genes for drought resistance improvement. On the basis of genetic dissection of droughtresistance of rice, more than 30 QTLs have been targeted for construction of near isogenic linesand marker-assisted molecular breeding. Several drought resistance-associated genes wereidentified through drought screening of T-DNA insertion mutants of rice. Hundreds of genesdifferentially involved in drought responses and adaptation were identified through comparativeexpression profiling analysis. More than 200 drought-responsive candidate genes weretransformed into rice for drought resistance testing, and a few genes (such as SNAC1, OsSKIPa,and OsLEA3-1) showed significant effect in improving drought resistance of transgenic rice.Finally, problems and perspectives of drought resistance improvement in rice will be discussed. 33
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 1: Molecular breeding for abiotic stress tolerancesHeat stress transcriptome analysis and Functional Characterization ofResponsive Genes in wheatQin DD1, 2, Peng HR1, 2, Ni ZF1, 2, Yao YY 1, 2, Zhou CL1, 2, Sun QX 1, 2, *1 Department of Plant Genetics & Breeding and State Key Laboratory for Agrobiotechnology, ChinaAgricultural University, Beijing100193, China2 Key Laboratory of Crop Heterosis and Utilization (MOE), Key Laboratory of Crop Genomics and GeneticImprovement (MOA) and Beijing Key Laboratory of Crop Genetic Improvement, China Agricultural University,Beijing100193, ChinaEmail: qxsun@cau.edu.cnWheat (Triticum aestivum L.) is a major crop around the world, and heat stress during the latestage affects its yield and quality badly. So, it’s urgent to elucidate the mechanisms of wheat heattolerance, and identify thermotolerance-related genes for future thermotolerant wheat breedingprogramme.In this study, using Affymetrix Genechip® Wheat Genome Array, we analyzed genome-widegene expression profiles of the leaves between two wheat genotypes with contrastingthermotolerance under heat treatment, namely, heat susceptible ‘Chinese Spring’ and heat tolerant‘TAM107’. A total of 6560 (~10.7%) probe sets were identified as heat responsive in our study.Except for heat shock proteins and heat shock factors, these genes also included transcriptionfactors, components involved in hormone biosynthesis and signaling, calcium signal pathway,RNA metabolism, primary and secondary metabolisms, as well as other stresses related proteins.Further analysis showed that, 313 probe sets were differentially regulated between the twogenotypes, 1314 were between heat treatments with and without pre-acclimation, while 4533between short and prolonged heat treatments. Furthermore, two genes, TaMBF1c (Multiproteinbridging factor 1, MBF1) and TaGAST (Gibberellin stimulated transcript), which were stronginduced by heat stress in both genotypes were cloned and functionally characterized.The complete ORF encoding TaMBF1c included 471bp, the deduced amino acid sequencerevealed existence of MBF1 and helix-turn-helix conserved domains at the N- and C-terminus,respectively, and was highly homologous to rice ERETC and AtMBF1c. TaMBF1c contained nointron in it. The 1074bp promoter region of it contained three heat shock elements (HSEs),identifying it as a potential heat shock factor regulated gene. Northern blot analyses showed thatthere was no detectable expression of TaMBF1c under control condition, and the expression of itwas rapidly and significantly induced by heat stress not only at seedling stage but also atflowering stage, and was only slightly induced by drought and H2O2 stresses, ABA and ACCapplication, however, not by rhythm, salt and MeJA treatments. In addition, ectopicover-expression of TaMBF1c in yeast imparts high temperature stress tolerance to wild type yeastcells. The most important is that thermotolerance was significantly increased in TaMBF1coverexpressed transgenic rice. 34
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Another heat-induced gene TaGAST was also gotten by in silico cloning and RT-PCR.Bioinformatic analysis showed that the sequence of TaGAST encoded a protein with 99 aminoacids which had a GASA domain in the C-terminal. In addition, the promoter region of TaGASTwas cloned using BD Genome Walker method, and HSE and several cis-elements involving inother abiotic stress response were found in this region.Consistently, the expression of TaGASTwas at low level in seedling leaves of the two wheat genotypes mentioned above, but stronglyinduced by stress factors, such as PEG, high salinity, oxidation and high temperature, and also thephytohormones such as ABA, ACC and MeJA treatment.The results suggested that this genemight be involved in various abiotic stress respons.In order to investigate the role of TaGAST inplant thermotolerance, it was over-expressed in Arabidopsis by Agrobacterium-mediatedtransformation method. The transgenic lines overexpressing TaGAST showed no phenotypicdifference compared to wild type under normal growth condition, but showedmembrane-thermostabler than WT. And had significantly higher survival rate under heat stress.All the above results indicate that these two genes have potential importance in improvingthermotolerance of wheat and other cereals, and the transgenic of wheat is underway. 35
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 2: Gene discovery and functionIdentification and application of the rice broad-spectrum blast resistance genePigmHe ZH*, Deng YWNational Key Laboratory of Plant Molecular Genetics, Institute of Plant Physiology and Ecology, ShanghaiInstitutes for biological Sciences, CAS, Shanghai 200032, China*E-mail zhhe@sibs.ac.cn).Rice blast is one of the most destructive diseases of rice. The identification and utilization ofbroad-spectrum resistance genes has been the most effective and economical approach to controlthe disease. A native Chinese variety, GM4, was identified with broad-spectrum and durableresistance. Genetic and mapping analysis indicated that blast resistance to nine isolates ofdifferent races in GM4 is controlled by the same dominant locus designated as Pigm, which wasidentified resistance gene cluster including 13 NBS-LRR members on chromosome 6 bymap-based cloning strategy, allelic to two known blast resistance genes Pi2 and Pi9. Sequencecomparison of the orthologous and paralogous genes between the Pigm/Pi9/Pi2 loci showed thatthe Pigm loci had undergone duplication result from LTR retrotransposon, unequal cross andillegitimate recombination during the evolution of the resistance gene cluster. Furthermore, ouranalysis showed that Pigm confers resistance to blast isolates from different cultivated regionsthan Pi9/Pi2/Pizt/Piz. In the Pigm locus, Pigm-1 controls leaf blast resistance, Pigm-2 confersneck blast which leads to large loss of grain yield. Genetic and transcriptional analysis suggestedthat broad-spectrum resistance might be attributed to the different expression patterns of diverse Rgenes. We have succeeded in developing elite hybrid rice lines with broad-spectrum blastresistance with molecular markers-assisted selection for Pigm, indicating good potential of thegene in rice molecular breeding. All the elite hybrid rice lines harboring the Pigm exhibited a highresistance or immunity to blast in natural blast nurseries nationwide from 2008 to 2010. 36
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 2: Gene discovery and functionMutant resources for functional studies of genes related to fertility in riceWu CYNational Key Laboratory of Crop Genetic Improvement and National Center of Plant Gene Research, HuazhongAgricultural University, Wuhan 430070, ChinaEmail: cywu@mail.hzau.edu.cnT-DNA tagging strategy is a high throughput approach for function analysis the plant genome.We have generated more than 100 thousand independent transgenic lines using the enhancer trapconstruct, consisting of the GAL4/VP16-UAS elements with GUS (or GFP) as the reporter. Thesystem has three built-in strategies for functional analysis of the rice genome. First, T-DNAinsertions cause gene mutations, providing an efficient approach for gene identification andisolation. Second, expression of the reporter gene indicates the presence of an enhancer element inthe neighboring genomic region, which can be used for isolation and characterization of theenhancer. Third, the lines showing spatial- or temporal-specific expression of the reporter genecan be used to drive ectopic expression of a transgene, thus useful for unveiling latent functions ofunknown or known genes.Employing our rice T-DNA insertional mutant library, we identified two genes, designed PAIR3and OsRPA1a, which play essential roles in DNA metabolism during meiosis process. Both pair3and Osrpa1a mutants exhibit a phenotype of completely sterile compared with their wild types.Genetic analysis of those mutants revealed that the T-DNA insertion tag co-segregated with thesterility phenotype. During meiotic prophase I, the pair3 mutant fails in homologous chromosomepairing and synapsis, resulting in no formation of bivalents and subsequent random segregation ofthe univalents in anaphase I. PAIR3 encodes a protein that contains putative coiled-coil motifs, butdoes not have any close homologs in other organisms. Primary results suggest that PAIR3 plays acrucial role in homologous chromosome pairing and synapsis in meiosis. Another mutant osrpa1aexhibits abnormal chromosomal fragmentation occurred in male meiocytes after anaphase I.Further study identified that the leaves of Osrpa1a were hypersensitive to DNA mutagens.Genetic complementation and RNAi results confirmed that OsRPA1a was responsible to themutant phenotypes in Osrpa1a. Our data suggest that OsRPA1a plays an essential role in DNArepair but may not participate in, or at least is dispensable for, DNA replication and homologousrecombination in rice. 37
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 2: Gene discovery and functionGene discovery from common wild rice (Oryza rufipogon Griff.)Sun CQState Key Laboratory of Plant Physilogy and Biochemistry, National Center for the Evaluation of AgriculturalWild Plants (Rice), China Agricultural University, Beijing 100193, P R China.Email: suncq@cau.edu.cnCommon wild rice (Oryza rufipogon Griff.), ancestor species of cultivated rice (O. sativa L.),constitute an important gene pool for rice improvement. To discover favorable genes from wildrice which have been lost or weakened in cultivated rice has become more and more important formodern breeding strategy. In recent years, we have developed two sets of introgression lines (ILs)derived from the cross between O. rufipogon from Jiangxi and Yunnan province of China, as thedonor, and elite cultivars, as the recipient. Several QTLs for yield-related traits, quality traits andtolerence to abiotic stress were mapped using introgression lines. Some major QTLs werefine-mapped and cloned. Two key genes, PROG1 and SHA1, controlling rice domestication wereidentified. PROG1 controlling prostrate growth of wild rice on chromosome 7 encodes a singleCys2-His2 zinc-finger protein. prog1 variants identified in O. sativa disrupt the prog1 functionand inactivate prog1 expression, leading to erect growth, greater grain number and higher grainyield in cultivated rice. SHA1 controlling seed shattering of wild rice on chromosome 4 encodes amember of the trihelix family of plant-specific transcription factors. The predicted amino acidsequence of SHA1 in wild rice is distinguished from that in cultivated rice by only a single aminoacid substitution (K79N) caused by a single nucleotide change (g237t). 38
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 2: Gene discovery and functionDiscovery of brown planthopper resistance gene in riceHe GCCollege of Life Sciences, Wuhan University, Wuhan 430072, ChinaEmail: gche@whu.edu.cnThe brown planthopper (Nilaparvata lugens Stal; BPH) is an insect that feeds on the leaf sheath ofrice (Oryza sativa L.) plants, ingesting nutrients specifically from the rice phloem using its styletmouthparts. In the last decade, the BPH has frequently caused widespread destruction of ricecrops and heavy losses of yields. The most economic and efficient method for controlling theBPH is to use the host resistance as part of IPM.To date, more than 19 BPH resistance genes in rice have been reported. Resistance of Bph1, bph2and Bph3 has been reported to be overcome by new biotypes of BPH. Wild rice germplasm is animportant gene pool for rice breeding. Two major loci for BPH resistance, Bph14 and Bph15,were detected in the F2 population and RIL population of Minghui63 X B5. Bph14 was mappedon the long arm of chromosome 3 and Bph15 on the short arm of chromosome 4. These loci werealso found to confer resistance to the white-backed planthopper.Analysis of recombination events in the Bph14 region delimited the gene to genomic segment of34-kb between SM1 and G1318. Two predicted genes encoding putative resistance proteins,designated Ra and Rb respectively, were identified after sequencing this region. Transgenicexperiment showed that Ra confers the resistance phenotype and is the Bph14 gene. The Bph14gene encodes a putative 1,323 amino acid protein containing a coiled-coil, nucleotide-binding andleucine-rich repeat (CC-NB-LRR) motif. Comparison analysis showed that in the LRR domain 54residues and two deletions of Bph14 were unique.Electronic penetration graphs (EPG) revealed that BPH insects spent more time walking, but lesstime ingesting phloem, on the plants carrying resistance genes Bph14 and Bph15 than they did onsusceptible plants. Tests with [14C]sucrose showed that insects ingested much less phloem sap onthe resistant plants than on susceptible plants. In the plants infested with the BPH, callose wasfound deposited on the sieve plates of the target sieve tubes, where the stylets had been inserted.Counts of the bright callose plugs revealed more callosic sieve plates in the resistant than insusceptible plants. Moreover, with prolonged BPH feeding, the callose deposition decreasedquickly in susceptible plants. It was found that the genes encode for callose decomposing enzymeβ-1,3-glucanase were differetially regulated in the resistant and susceptible rice plants. In thesusceptible rice the β-1,3-glucanase gene Osg1 and Gns5 were enhanced, and thereby facilitatedthe BPH’s continued feeding from the phloem in the susceptible plants, while in the resistantplants, these genes expression unchanged. As a result, BPH feeding on the resistant rice plantswere suppresed. 39
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 2: Gene discovery and functionMolecular basis of cytoplasmic male sterility in riceWang Z, Zou Y, Luo D, Liu Z, Xu H, Wu H, Guo J, Zhang Q, Ye S, Chen Y, Liu YG*Key Laboratory of Plant Functional Genomics and Biotechnology of Education Department, Guangdong Province,College of Life Sciences, South China Agricultural University, Guangzhou 510642, China.*Email: ygliu@scau.edu.cnThe successful breeding and commercial cultivation of hybrid rice is one of the most importantachievements in agriculture. Hybrid rice has been developed and released in 1970s in China, whichhas about 20% yield advantage over improved inbred varieties. Since the late of 1980’s, hybrid ricehas occupied ~55% (~15-17 million hectares) of the total rice planting area each year in China.Therefore, hybrid rice has contributed tremendously to the food security in China, and given a greatimpact to agriculture. The successful development of hybrid rice is mainly due to the developmentand utilization of cytoplasmic male sterility (CMS) systems. Three types of CMS systems,Wild-abotive (WA), Boro II (BT), and Hong-Lian (HL), have been used for the hybrid breeding.To reveal the molecular basis of the cytoplasmic male sterility systems in rice, we have identifiedand functionally studied the genes conferring the CMS and restoration. We found that amitochondrial open reading frame of previously unknown function in Boro II cytoplasm, orf79,encodes a cytotoxic peptide that causes the male sterility. Furthermore, we isolated two restorergenes, Rf1a and Rf1b, at the previously reported single locus Rf1, revealing that Rf1 is a complexlocus. Rf1a and Rf1b encode PPR (Pentatrico Peptide Repeat) proteins, and they target tomitochondria to cleave and degrade the orf79 mRNA, respectively, thus silence orf79 and restorethe mal fertility. When both restorers are present in the hybrids, Rf1a preferentially cleave the orf79mRNA, showing an epistatic effect over Rf1b. The study further revealed that Rf1a has a role topromote the editing of the mitochondrial atp6 mRNA, suggesting that this may be its primaryfunction, while the action as the fertility restorer be a new function.CMS-WA is the most widely used system for hybrid rice. We identified a novel mitochondrial geneconferring CMS-WA. Transformation of rice and Arabidopsis with this gene caused male sterility.CMS-WA is restored by Rf loci, Rf3 and Rf4, via suppressing the function of this CMS gene withdifferent mechanisms. Evolutionary analysis revealed that this CMS gene was generated throughrearrangement of multiple fragments of the mitochondrial genomes and unknown sources in thislocus during the evolution of wild rice species. Further, we studied the molecular mechanism of theCMS induction involving in the cytoplasmic-nuclear interaction. 40
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 2: Gene discovery and functionToward map-based cloning of a good eating-quality QTL derived from an eliteJapanese rice cultivar KoshihikariHori K1*, Takeuchi Y2, Nagasaki H1, Ando I2, Yano M11 National Institute of Agrobiological Sciences, 2-1-2 Kannondai, Tsukuba, Ibaraki 305-8602, Japan2 National Institute of Crop Science, 2-1-18 Kannondai, Tsukuba, Ibaraki 305-8518, Japan*E-mail: horikiyo@affrc.go.jpEating quality is an important trait to consider in rice breeding, because it determines consumerpreference and the rice price. The eating quality of cooked rice is a complex trait determined bymultiple genes and is largely affected by environmental factors. Although several physicochemicalproperties of the rice grain, such as amylose and protein contents, pasting properties and gelconsistency are used to evaluate eating quality, a sensory test of cooked rice is still required forthe final selection procedure in rice breeding. The sensory test is time-consuming andlabor-intensive because trained panels evaluate each breeding line for appearance, taste, andtexture of the cooked rice by eating it. A japonica rice cultivar Koshihikari has a good eatingquality including high glossiness, a high level of stickiness, good taste, and low hardness ofcooked rice. We evaluated the eating quality of cooked rice using the sensory test in a set ofreciprocal backcrossed inbred lines (BILs) from crosses between Nipponbare and Koshihikari in2006 and 2007. The major quantitative trait loci (QTL) for eating quality were detected on theshort arm of chromosome 3 in the two BILs. The Koshihikari allele of the QTL increased eatingquality. To validate the eating quality QTL, we developed a substitution line with a Koshihikarisegment on the short arm of chromosome 3 in a Nipponbare genetic background, and evaluatedthe eating quality of the substitution line using sensory tests in 2008 and 2009. The eating qualityof the substitution line was improved as compared with Nipponbare in both seasons. In order toscreen for putative candidate genes of the eating quality QTL, a large chromosome segment (11.3Mbp) of the genome was sequenced. Sequence comparison between Nipponbare and Koshihikarirevealed insertion/deletion polymorphisms and single nucleotide polymorphisms in the sequencesof 13 predicted genes in the candidate region of the QTL. RT-PCR revealed that nine of the 13genes were expressed in the endosperm during the ripening period after pollination. Forfine-mapping of the eating quality QTL, we developed additional substitution lines to replacedifferent Koshihikari segments on the short arm of chromosome 3 in the Nipponbare background.Sensory tests of these substitution lines are now underway to narrow down the candidate regionfor the eating quality QTL. 41
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 2: Gene discovery and functionMap-based cloning of QTL genes for flowering time/maturity in soybeanXia ZJ1,2*, Watanabe S2, Liu B1, Tsubokura Y2, Harada K21 Northeast Institute of Geography and Agroecology, CAS, Harbin 150081, China;2 National Institute of Agrobiological Sciences, 2-1-2 Kannondai, Tsukuba, Ibaraki, 305-8602, Japan.* Email: xiazhj@neigaehrb.ac.cnFlowering represents the transition from the vegetative to the reproductive phase in plants.Various external cues, such as photoperiod and temperature, are known to initiate plant floweringunder the appropriate seasonal conditions. In soybean [Glycine max (L.) Merrill], several maturityloci, designated as E loci have been characterized by classical methods. Of these E loci (E1 to E7),the E1, E3, and E4 loci have been suggested to be related to photoperiod sensitivity under variouslight conditions. Successful cloning of these genes is very important to understand the floweringgene network in soybean. As well, deciphering these identities can help us to understand thenatural evolution and domestication, and impose very important impact on soybean breeding andproduction. As the draft soybean sequence has been made publically available, it provides us greatgenetic resources, especially for positional cloning. As clonings of E3 (Watanabe et al. Genetics.182, 1251-1262) and E4 (Liu et al. Genetics 180: 995–1007) have been accomplished, we arefocused on isolation of E2 and E1 loci. Map-based cloning strategy using residual heterozygouslines was applied to the isolation of the gene responsible for these two loci. One candidate geneconsisting of 1170 amino-acid was identified for E2 locus. Common premature stop codon at thetenth exon (521aa) was observed in Misuzudaizu allele and other NILs originated from Harosoy(e2/e2; PI548573). Furthermore, a mutant line harboring another premature stop codon (735aa)showed early flowering phenotype than original variety Bay (E2/E2; PI553043). In contrast toother plant species, null mutation of E2 gene in soybean did not show any notable deleteriouseffects on the phenotype and can be a good genetic resource to control maturity in widegeographic area in soybean breeding program. Most importantly, we are cloning E1/FT1 using apopulation derived from a cross between two Harosoy NILs. With identified recombinants, wehave successfully narrowed down E1 locus from about 300 kb to 17 kb, where a single candidategene was identified. Further functional confirmation is in progress. With these four majorflowering time genes identified, the flowering time gene networks in soybean will be greatlyunderstood. 42
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 3: Molecular breeding for biotic stressesFrom QTLs for disease resistance to marker-assisted selection in durum wheatMaccaferri M1*, Sanguineti MC1, Bassi F1, Mantovani P1, Terracciano I1, Ammar K2, Massi A3, Chen F4, BiniF1, Ratti C1, Rubies-Autonell C1, Vallega V5, Simkova H6, Kolmer J7, Kema G8, Keller B9, Czembor J10,Tuberosa R11 Dept. of Agroenvironmental Science & Technology (DiSTA), U. of Bologna, 40127 Bologna, Italy; 2CIMMYT, 56134Texcoco, Mexico; 3Società Produttori Sementi Bologna (PSB), 40050 Argelato (BO), Italy; 4Institute of Crop Science,CAAS, Beijing 100081, China; 5CRA Experimental Institute for Cereal Research, 00191 Rome, Italy; 6Institute ofExperimental Botany, CZ-77200 Olomouc, Czech Republic; 7Cereal Disease Laboratory, USDA-ARS, St. Paul, MN55108, USA; 8Plant Research International (PRI), Wageningen, 6700 AA, The Netherlands; 9Institute of PlantBiology U. of Zurich, 8008 Zurich, Switzerland; 10Dept. of Genetics & Plant Breeding, IHAR, 05870 Blonie, Poland*E-mail: marco.maccaferri@unibo.itDurum wheat (Triticum durum Desf.) is a main cereal crop for the Mediterranean area where production ischallenged by fungal pathogens (e.g. leaf rusts, powdery mildew, Fusarium, etc.) and in some areas, also bySoil-Borne Cereal Mosaic Virus (SBCMV). The identification of effective sources of resistance, their geneticmapping and the development of molecular markers suitable for marker-assisted selection are thus majorobjectives of durum wheat improvement. The positional cloning of the relevant loci would also be of greatapplicative value.In collaboration with CIMMYT and plant pathologists, we have undertaken a program for identifying andmapping useful genetic variation through a joint linkage and association mapping approach. The developmentand phenotypic characterization of two RIL populations (from Meridiano × Claudio and Colosseo × Lloyd),each including ca. 180 RILs, allowed us to identify major genes/QTLs for: (i) leaf rust resistance on chr. 7BL(C×L; Maccaferri et al., 2008; TAG 117: 1225–1240; Mol. Breed. 2010, DOI 10.1007/s11032-009-9353-0.), (ii)powdery mildew resistance (M×C) on chrs. 6BL and 7BL, and (iii) SBCMV resistance (M×C) on chr. 2BS.Association mapping with a panel of 210 elite accessions has been used to validate and to further dissect thesemajor resistance loci. A fine-mapping effort is underway for the target loci. The analysis of rice-wheat syntenyprovides the bases for SNP development and candidate gene identification. Based on the obtained results,marker-assisted selection is being implemented in collaboration with the seed company Produttori SementiBologna. First, the proprietary germplasm has been haplotyped at the major genes/QTLs in order to identify thevaluable lines that already harbor the favourable resistance alleles. Molecular selection is currently being usedto screen segregant F2:4 breeding populations. High-throughput, PCR-based codominat markers have beendeveloped (BIOEXPLOIT project) for MAS activities. 43
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 3: Molecular breeding for biotic stressesGenomic approaches to plant defense research and crop improvement forinsect resistanceHuang YHUSDA-ARS Plant Science Research Laboratory, 1301 N. Western Road, Stillwater, OK 74075 and Departmentof Botany, Oklahoma State University, Stillwater, Oklahoma 74078, USAEmail: yinghua.huang@ars.usda.govPlant-insect interaction is a complex and dynamic process, leading to a variety of beneficial anddeleterious outcome. Mechanisms of plant defense against attack by insect pests, includingconstitutive and induced defenses, have been evolving for millions of years and are thereforeshared across many plant families. Understanding of the interactions between two kingdoms andthe defense mechanisms operating in host plants, such as the insect genes required for attackingand the plant genes responsible for defense, can be utilized to design crops with enhancedresistance.Research in life sciences at the post-genomics age is currently undergoing a dramatictransformation, and genomic approaches are beginning to revolutionize our understanding of plantdefense. These sophisticated genomic tools, including genome sequencing, gene expressionprofiling, functional genomics, marker-assisted selection, genetic transformation etc., are alreadyapplied to examine the complex process of plant and pest interactions. Genome sequencingprojects of a few crops, including rice and sorghum, have been completed. The resulted geneticblueprints provided by DNA sequences will allow the identification of genes involved in hostdefense and regulatory factors that control the induction of resistance genes. Molecular mappinghas permitted locating chromosomal regions harboring the resistance to insects and identifyinggenetic markers enabling more efficient plant breeding or genetic manipulation. Transgenic plantsalready have been an unimagined tools and capacities in determination of gene functions inplant-insect interactions and in pursuit of molecular breeding for insect resistance.These successful samples highlighted the recent progress in the studies of plant-insect interactionsand provided valuable data on the genome structure of host plants and the diversity of defensemechanisms used by plants to protect themselves from insects. However, our understanding ofgene function remains behind the pace of genome sequencing and gene discovery. Over the nextten years, further progress and new breakthroughs are anticipated through integrated biologystudies and the use of newly genomics tools; then the resulting new knowledge will lead to thedevelopment of more durable insect resistance for crop plants and novel strategies for crop pestmanagement. 44
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 3: Molecular breeding for biotic stressesImprovement of maize resistance to head smut and stalk rotXu MLNational Maize Improvement Center of China, China Agricultural University, 2 west Yuanmingyuan Road,Beijing, 100193, ChinaStalk rot and head smut are among the most destructive diseases in maize production in China.Disease control by using fungicide has been proved inefficient due to soil-borne infection, while,genetic strengthening of maize against stalk rot and head smut is the most cost-effective andenvironmentally friendly way to curtail the spread of the diseases.Highly resistant maize inbred lines, ‘Ji1037’ against head smut and ‘1145’ against stalk rot, havebeen screened from maize germplasm and used to prepare mapping populations for QTL analyses.The individuals with resistance QTL were backcrossed to the susceptible parent to generate nextbackcross generation. This process has been repeatedly conducted to produce advanced backcrosspopulations. In each backcross generation, recombinants were selected and then backcrossed tothe susceptible parent to generate their backcross progeny. A progeny testing was introduced todeduce phenotype of parental recombinant. If the progeny with the donor region are moreresistant than those without the donor region, indicating the presence of resistance QTL, orotherwise, no any resistance QTL was present in the parental recombinant. Analysis of bothresistance and donor region for every recombinant allowed fine-mapping of the resistance QTL.With this approach, the major resistance QTL to stalk rot and head smut has been mapped intointervals of 170kb and 190kb, respectively. The markers in the mapped regions were used toscreen two BAC libraries constructed from the inbred lines ‘1145’ and ‘Huangzao 4’ to build BACcontigs covering the resistance QTLs. The minimal tilling BACs were subjected to sequencingand gene prediction. By comparison of the predicted genes among ‘1145’, ‘Huangzao 4’ and B73(whole genome sequence is publicly available) allowed identification of candidates for resistanceQTLs. The candidate resistance genes, three for each disease, were then cloned into expressionand RNAi vectors for functional validation via genetic transformation.The makers developed on or closely-linked to the resistance genes have been extensively used formarker-assisted selection to genetic improvement of maize resistance to the diseases. This hasbeen proved very successful since highly resistant to head smut and stalk rot has been observed inten improved inbred lines and a number of isogenic lines, respectively. We have collaborated withmany institutions in China to accelerate application of this marker technology to strengthen maizeresistance to both head smut and stalk rot. 45
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 3: Molecular breeding for biotic stressesEnhancing broad spectrum resistance to rice diseasesWang SPNational Key Laboratory of Crop Genetic Improvement, National Center of Plant Gene Research, HuazhongAgricultural University, Wuhan 430070, ChinaEmail: swang@mail.hzau.edu.cnBroad-spectrum resistance refers to resistance against two or more types of pathogen species orthe majority of races of the same pathogen species. Bacterial blight caused by Xanthomonasoryzae pv. oryzae (Xoo), bacterial streak caused by X. oryzae pv. oryzicola (Xoc), and fungal blastcaused by Magnaporthe grisea are devastating diseases of rice worldwide. Application of hostresistance to these pathogens is the most economical and environment-friendly approach to solvethis problem. Quantitative resistance conferred by quantitative trait loci (QTLs) is presumablyvaluable sources for broad-spectrum disease resistance. However, molecular evidence thatsupports this hypothesis is rare. Resistance QTLs are also only identified genetic resources fromrice against Xoc. Although a large number of QTLs against these rice diseases have beenidentified, these sources have not been used effectively in rice improvement because of thecomplex genetic control of quantitative resistance and because of the genes underlying most ofresistance QTLs being unknown. We have characterized several minor resistance QTLs in rice bythe strategy of validation and functional analysis of the QTL. Functional characterization of theencoding proteins of these resistance QTLs indicates that these genes belong todefense-responsive genes group. These QTLs can mediate broad-spectrum resistance and can beused in breeding programs by manipulating their expression. 46
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 3: Molecular breeding for biotic stressesMolecular mapping of adult-plant resistance genes to stripe rust and powderymildew and validation of allelic specific markers for Lr34/Yr18/Pm38 inChinese wheat cultivarsXia XC1,*, He ZH1, 2, Lan CX1, Liang SS1, Ni XW1, Li ZF11 Institute of Crop Science/National Wheat Improvement Center, Chinese Academy of Agricultural Sciences(CAAS), 12 Zhongguancun South Street 100081, Beijing, China; 2International Maize and Wheat ImprovementCenter (CIMMYT) China Office, c/o CAAS, 12 Zhongguancun South Street, Beijing 100081, China.*Email: xiaxianchun@caas.net.cnStripe rust and powdery mildew, caused by Puccinia striiformis f. sp. tritici (PST) and Blumeriagraminis f. sp. tritici, respectively, are devastating wheat diseases in China. The 145 Chinesewheat cultivars and advanced lines from the autumn-sown wheat regions were investigated.Ninety-eight lines of them were tested for seedling response to 26 PST races in the greenhouse,and 135 were tested for adult-plant resistance (APR) in the field. The results indicated that genesYr2, Yr3a, Yr4a, Yr6, Yr7, Yr9, Yr26, Yr27, and YrSD, either singly or in combinations, werepostulated in 72 lines, whereas known resistance genes were not identified in the other 26accessions. The resistance genes Yr9 and Yr26 were found in 42 and 19 accessions, respectively;and 33 cultivars and advanced lines showed APR to stripe rust at two locations in both seasons.Using the allele-specific markers (cssfr1–cssfr5) for Lr34/Yr18/Pm38, we tested 231 Chineseimproved wheat cultivars and 422 landraces. The resistance-allele specific PCR fragments weredetected in 14 improved wheat cultivars, with a frequency of 6.1%, whereas these fragments wereamplified in 359 landraces, with a frequency of 85.1%. However, about 25% of these landraceswith resistance-allele specific PCR fragments showed highly susceptible to stripe rust. DNAsequencing for them didn’t find any mutations in the coding region of the gene, and theirsusceptibility might be resulted from the mutation in the promoter region or the presence of aninhibitor of the gene Lr34/Yr18/Pm38. A total of 540 simple sequence repeat (SSR) markers were screened to map quantitative traitloci (QTLs) for APR to stripe rust in a doubled haploid (DH) population of 137 lines derived fromthe cross Pingyuan 50× Mingxian 169. The DH lines were planted in randomized complete blockswith three replicates in Gansu and Sichuan provinces during the 2005-06, 2006-07, and 2007-08cropping seasons, providing data for four environments. Artificial inoculations were carried out inGansu and Sichuan with the prevalent Chinese PST race CYR32. Inclusive composite intervalmapping (ICIM) detected three QTLs for APR to stripe rust on chromosomes 2BS, 5AL, and 6BS,designated QYr.caas-2BS, QYr.caas-5AL, and QYr.caas-6BS, respectively, explaining from 4.5 to19.9% of the phenotypic variance.The 406 SSR markers were used to map QTLs for APR to powdery mildew in a DH populationwith 181 lines derived from the cross Bainong 64×Jingshuang 16. With the method of composite 47
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京interval mapping, four QTLs for APR to powdery mildew were detected on chromosomes 1A,4DL, 6BS and 7A, designated QPm.caas-1A, QPm.caas-4DL, QPm.caas-6BS and QPm.caas-7A,respectively, explaining from 6.3 to 22.7% of the phenotypic variance. Two QTLs QPm.caas-4Dand QPm.caas-6BS were stably detected across different environments with a high genetic effecton powdery mildew resistance, accounting for 15.2 to 22.7% and 9.0 to 13.2% of phenotypicvariance, respectively. A population of 200 F3 lines from the cross Lumai 21×Jingshuang 16 was tested for powderymildew reaction in Beijing and Anyang in the 2005-06 and 2006-07 cropping seasons, providingdata for 4 environments. A total of 1,375 SSR markers were screened for associations withpowdery mildew reactions. Three QTLs for APR to powdery mildew were detected by ICIM.These were designated QPm.caas-2BS, QPm.caas-2BL and QPm.caas-2DL, respectively, andexplained from 5.4 to 20.6% of the phenotypic variance across 4 environments. QPm.caas-2BSand QPm.caas-2DL were likely new QTLs for APR to powdery mildew, flanked by SSR markersXbarc98 - Xbarc1147 and Xwmc18 - Xcfd233, respectively. 48
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 3: Molecular breeding for biotic stressesTransmission and molecular characteristics of Southern rice black-streakeddwarf virus, a new Fijivirus threating rice production in AsiaZhou GH*, Wang Q, Pu LL, Zhang SGLaboratory of Plant Virology, South China Agricultural University, Guangzhou, China* Email: ghzhou@scau.edu.cnSouthern Rice Black Streak Dwarf Virus (SRBSDV, genus Fijivirus, Family Reoviridae), a newvirus infecting rice discovered in 2001 in Guangdong province, spread raphidly throughoutsouthern China and northern Vietnam, and became one of the most important virus threating riceproduction in Southeast Asia. The virus was transmitted by the white back planthopper (WBPH,Sogatella furcifera, Hemiptera: Delphacidae), a long-distance migration rice pest, in a persistentmanner. The virus replicates in the WBPH body, but can not be transmitted to offspring throughegg. About 60% WBPH individual nymph at 4-5th instar stage was viruliferous in the populationdeveloped on the infected rice plant. Both nymph and adult transmit the virus while the formerwith higher effieciency. Acquisition and inoculation peorids were at least 10-30 minutes, andincubation peorid were 5-7 day when the virus was artificially transmitted through WBPH nymph.The complete genome sequences of this virus, containing ten linear segments of double-strandedRNA, were determined. Based on the nucleotide identity analysis, SRBSDV was most closelyrelated to MRDV and RBSDV. Comparsing to RBSDV, S1, S2 and S10 were most conserved withidentities of 78.5-79.2% while S5 and S6 were the least conserved with 70.6-71.6% identities. Noreconbination was found between this virus with related viruses. Transmission and molecularcharacteristics revealed in this study provided a base for virus resistance screening and breeding,especially by molecular strategy. 49
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 4: New transgenic technologies, products and marketsNew Transgenic TechnologiesRichard BroglieDirector, DuPont Agricultural BiotechnologyMethods to insert genes at defined, previously characterized genetic loci are highly desirablemolecular tools in plant biotechnology for excision and removal of marker genes and forcontrolled regulation of foreign gene expression. Homologous recombination andDNA-recombinase-mediated site-specific integration are promising technologies to accomplishthis. This paper will describe the successful targeting of transgenes to predefined sites insoybean and corn using the yeast FLP-FRT recombination system. Experiments will also bepresented which demonstrate the feasibility of using re-designed homing endonucleases tointroduce targeted mutations in crop plants. 50
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 4: New transgenic technologies, products and marketsSimultaneously changing several quality traits of Brassica napus by onetransgenic eventPeng Q, Hu Y, Wei R, Zhang Y, Guan C, Ruan Y, Liu CL*Hunan Provincial Key Laboratory of Crop Germplasm Innovation and Utilization, Hunan Provincial Institute ofOil Crops, Hunan Agricultural University, Changsha 410128, ChinaDevelopment Biology Department, Life Science and Technology College, Hunan Agricultural University,Changsha 410128, China* Email: liucl100@126.comThe fatty acid composition in the seed oil was significantly modified following the introduction oftransgenes. To further enhance the desirable characteristics of rapeseed oil, it would be beneficialto develop a new approach for the simultaneous silencing of two or more target genes. Our goalsin the current study were to 1) increase oleic acid to more than 75%, 2) reduce polyunsaturatedfatty acids (PUFA) to about 10% and erucic acid to zero, and 3) accomplish these changes in asingle-transformation event. In a single transformation, two fragments amplified from the fattyacid 12-desaturase 2 (BnaFAD2) and fatty acid elongase 1 (BnaFAE1) genes of Brassica napuswere linked together to form a fusion fragment. The fusion fragment was then used to assembleunique intron-spliced hairpin interfering constructs. In the transgenic plant FFRP4-4, theexpression of BnaFAD2 and BnaFAE1 genes was completely inhibited. The composition of oleicacid in FFRP4-4 rose to 85%, PUFA dropped to 10% and erucic acid was undetectable. All hybridF1 seeds obtained from the reciprocal crossing of FFRP4-4 and GX-parents (with different geneticbackgrounds) contained more than 80% oleic acid, about 10% PUFA and very low, orundetectable, erucic acid. The results confirmed that the fusion fragment silencing construct cansimultaneously and effectively silence the target genes on a consistent basis. The strategy providesa useful tool for detecting gene function and advancing genetic engineering techniques for theimprovement of agricultural crops. 51
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 4: New transgenic technologies, products and marketsIn situ Pistil Delivery: A High Throughput Method of Brassica GeneticTransformationGuo XL1, Dong CH1, Kurup S2, King G2, Shi L1, Cai L3, Fu H4, Wang Z1, Wang HZ1, Liu SY1*1 Oil Crops Research Institute of Chinese Academy of Agriculture Sciences and Key Laboratory of Oil CropBiology, Ministry of Agriculture, No. 2 Xudong Road 2, Wuhan, Hubei, China, 4300622 Rothamsted Research, Harpenden, Herts, AL5 2JQ, UK. 3College of Plant Science and Technology, HuazhongAgricultural University, No.1 Shizishan Street, Wuhan, Hubei, China, 430070. 4Ningbo Institute of Technology,Zhejiang University, No. 1 Qianhu South Street, Ningbo, Zhejiang, China, 315100.*Email: liusy@oilcrops.cn,Conventional Agrobacterium-mediated transformation methods rely on tissue culture and explantsregeneration. Here we described a new high throughput method without tissue culture oradditional equipment for producing transgenic Brassica plants. This method of in situ pistildelivery involved placement of a liquid drop containing binary vector DNA on the cut surface ofpollinated Brassica stigmas. Transformants, as Brassica napus L. cv Zhong Shuang 9 (ZS9) forexample, could be obtained from pistils pollinated only within 24 hours, indicating the timing oftransformation was crucial to the efficiency. High transformation frequency (Basta-resistantseedlings per 100 pistils) of 4.1% was achieved when ZS9 pistils were transformed at 18 hourspost-pollination. The method was validated by introducing different binary vectors into sevenother B. napus, two B. rapa and three B. oleracea lines/genotypes. The transformation frequenciesranged from 0.4% to 3.1% in B. napus, 10.3% to 16.3% in B. rapa and 0.7% to 2.7% in B.oleracea tested lines. Transgenic events were confirmed by PCR, β-glucuronidase (GUS)histochemical assay and Southern blots. A typical 3:1 (Basta-resistant: Basta-sensitive)segregation ratio is indicated in transgenic ZS9 subsequent generations, with no cytoplasmiceffect detected after reciprocal hybridization between transgenic lines and the wild type. Itappeared likely that gene delivery occurs after double fertilization for both embryo andendosperm can express the foreign GUS gene. Moreover, there are co-segregation evidencesdemonstrating the potential of this in situ pistil delivery method for mutagenesis and transgenicbreeding. 52
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 4: New transgenic technologies, products and marketsWheat genetic transformation in China -current status and future prospectsXia LQ 1*, He Y 1, Liang H 2, Wang DW 2, Ma YZ 1, Jones H 31 Institute of Crop Sciences /The National Key Facility for Crop Gene Resources and Genetic Improvement,Chinese Academy of Agricultural Sciences (CAAS), 12 Zhongguancun South Street, Beijing 100081, China.2 Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.3 Department of Plant Science, Rothamsted Research, Harpenden, AL5 2JQ, UK.* Email: xialq@mail.caas.net.cnGenetic transformation is fundamental to wheat functional genomics and improvement throughgenetic engineering. At the present time, biolistic and Agrobacterium-mediated transformation aretwo mainly employed ways for foreign gene transfer into wheat in China, although other DNAdelivery methods were also tried with varying degree of success. Agrobacterium-mediatedtransformation is perceived to have several advantages over other forms of transformation (suchas biolistic), including the ability to transfer large segments of DNA with minimal rearrangement,lower transgene copy number, low cost and may facilitate the removal of plant selectable markergenes by segregation, when the presence of unnecessary DNA and transgene arrangement/copynumber are scrutinized as part of the regulatory processes. Cereal species, particularly wheat(Triticum aestivum L.), have lagged behind dicots in their ability to be transformed byAgrobacterium and remain to be genotype-dependent, although there has also been a big progressin recent years, but it has been confined mainly to a few responsive varieties with quite differenttransformation frequencies.Here we not only review the current status of wheat genetic transformation in China, but alsoreport the successful Agrobacterium-mediated transformation of Chinese wheat varieties anddurum wheat cv Stewart by using the pGreen/pSoup system with improved transformationfrequency, in which the effect of the concentration of acetosyringone and auxin in the inoculation,co-cultivation and induction media on T-DNA transfer and regeneration efficiency werethoroughly investigated. Furthermore, the research and development (R&D) initiative ongenetically modified (GM) plants in China provides an opportunity and necessarily accentuatesthe development of a safe, precise and effective wheat genetic transformation system for latercommercialization at the same time. 53
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 4: New transgenic technologies, products and marketsA new effective selection marker for crop transformationLiu JH, Qiao L, Kong XF, Wang LY, Zhou JL, Xia M*National Center for Molecular Crop Design, Beijing, ChinaSelectable marker genes are very important for the crop transformation technologies because theeffective selection marker genes allow us to identify and select the plants with the integrated DNAand to monitor the transformed progeny. Paraquat (methyl viologen)-resistant gene, pqr-5,isolated from bacteria Ochrobactrum anthropi KT-q077, was successfully used as an efficientselection marker for the development of transgenic plants in Arabidopsis, rice, corn, soybean andcotton. Transgenic plants resistant to paraquat were positive upon PCR and RT-PCR by pqr-5gene-specific primers. The plants without expression of pqr-5 gene were killed quickly byparaquat, while the transformed plants were resistant to paraquat with the recommendedconcentration as a weedkiller. This selection strategy is faster, clearer and more visible in contrastto some traditional selections. The existence of paraquat-resistant marker gene in transgenic cropscontain useful novel traits while marker genes usually are not needed once transgenic plants havebeen identified. Therefore, the lack of phenotype variation, characterization of paraquat herbicideand low cost of paraquat makes this selection marker an attractive alternative for the croptransformation. 54
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 4: New transgenic technologies, products and marketsEnhancing the Lysine in wheat grain by genetic transformation of a Lysine richprotein gene CflrMa HX*, Sun XB, Fang R, Xu L, Zhang P, Yu GH and Zhang XInstitute of Biotechnology, Jiangsu Academy of Agricultural Sciences* Email: hxma@jaas.ac.cnWheat is the major source of protein for the nutrition of humans and livestock, but is deficient incertain essential amino acids when used as food for monogastic animals. In particular, theycontain low levels of lysine (the first limiting amino acid) resulting from deficiencies of thisamino acids in the prolamin storage proteins, which account for about half of the total nitrogen inthe mature grains. Genetic engineering has great potential for increasing the essential amino acidcontent of wheat grains. Expression of additional genes for lysine-rich proteins may lead toincreased accumulation of lysine in proteins.To obtain a new lysine-rich protein gene from species of Solanaceae, a 390bp fragment wasamplified from pepper (Capsicum frutescens) cultivar ‘Jiangshu 7’ by using RT-PCR with itsmature pollen cDNA as the template and the conservative sequences of potato and tomatolysine-rich protein genes as the primers. A full-length cDNA with completed open reading frameof 223 amino acids was cloned by using the strategy of RACE (rapid amplification of cDNA ends).This cDNA was designated as Cflr (Capsicum frutescens Lysine Rich), which contained 920 bpwith an untranslated region of 84 bp at 5’ end and a polyA tail at the 3’ end. BLAST searchagainst NCBI showed that the Cflr gene shared 50% -60% identity with the lysine-rich proteingenes from potato and tomato in nucleotide and 40% -50% in amino acid. The lysine content ofCFLR protein was 21·2%, which was the highest in the reported natural Lysine-rich proteins, andthe Threonine content was also high with the content at 10·3%. Analysis of semi-quantitativeRT-PCR indicated that Cflr gene was transcribed in mature pollen and petal largely, less in leafand hardly in immature anther, stem and root.The Lysine rich protein gene was transformed to the immature embryo in wheat cultivar Yangmai12 by using gold particle bombardment with the expression vector being constructed withlysine-rich protein gene Cflr from pepper, the GluB-1 promoter from rice and the Bar gene forscreening. The resistant calli differentiated to seedlings and 114 transgenic plants were obtained.PCR testing and Southern blot showed that Lysine rich protein gene had been transformed to thegenome of Yangmai 12, and it could be stably expressed in its next generation. Analysis of realtime PCR and semi-quantitative RT-PCR indicated that the Cflr gene had been expressed in T1transgenic wheat cultivar Yangmai 12, and expression abundance of transgenic plants weredifferent. The content of protein and lysine in T2 plant seeds of 81 transgenic plant lines wereanalyzed. The test results showed that lysine content in grains of transgenic wheat was rangedfrom 0.32% to 0.97% with the average of 0.55% improved 71% compared to control. 55
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 4: New transgenic technologies, products and marketsTransgenic strategies for improving drought tolerance traits in chickpeaBhatnagar-Mathur P, Krithika A, Rao JS, Vadez V, and Sharma KKInternational Crops Research Institute for Semi-Arid Tropics (ICRISAT), Patancheru, Andhra Pradesh,Patancheru, AP 502 324Water deficit is the most prominent abiotic stress that severely limits crop yields, thereby reducingopportunities to improve livelihoods of poor farmers in the semi-arid tropics (SAT) where most ofchickpea cultivation is done. In chickpea, annual losses of over 3.7 million tones have beenestimated to be due to water deficit conditions alone. Sustained long-term efforts in developingchickpeas with better drought tolerance through conventional breeding have been met with onlylimited success mainly because of an insufficient understanding of the underlying physiologicalmechanisms and lack of sufficient polymorphism for drought tolerance-related traits. At ICRISATwe have used transgenic technology to speed the process of molecular introgression of stressresponsive genes such as P5CSF129A and DREB1A in chickpea. Over 50 transgenic events with35S:P5CSF129A and 18 independently transformed plants of rd29A:DREB1A were developed andanalyzed at molecular and physiological levels. The putative transformants were confirmed for thepresence and expression of the transgenes by using PCR, RT-PCR, semi-quantitative RT-PCR andSouthern blot analysis. The evaluation of the transgenic chickpea plants was done underprogressive water stress conditions to understand the physiological effect of the inserted genes atthe whole plant level. The transgenic events carrying the P5CSF129A and DREB1A genes werephenotyped under both well-watered and water-limited conditions in greenhouse based ontranspiration efficiency (biomass produced per kg of water), photosynthetic activity, stomatalconductance and root length. Several events had superior transpiration efficiency, photosyntheticactivity, stomatal conductance and total transpiration under water limited conditions incomparison to the untransformed parent. All the selected transgenic events had a transpirationdecline upon soil drying in drier soil than in the untransformed controls. Since it is critical toassess how these putative differences in the transpiration pattern under water deficit conditionsrelate to the component traits of yield. The focus is on phenotyping different transgenic events inconditions close to field like conditions using a trait-based approach dissecting yield intocomponents under water stress. The current status and future plans on engineering droughttolerance in chickpea will be discussed. 56
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 4: New transgenic technologies, products and marketsIdentification of stress-inducible and tissue-specific promoters in riceLi ZX, Liu M, Li XJ, Xia M, Zhou JL*National Center for Molecular Crop Design, Beijing, ChinaFor monocotyledonous crops transformation, there is a shortage of efficient promoters forstress-inducible or tissue-specific expression of transgene. Here, we report isolation andidentification of several drought-inducible or tissue-specific promoters in rice. We selectedcandidate genes from the available microarray data sets and then did RT-PCR assay for all genes.Promoters of the genes with stress-inducible or tissue-specific expression pattern were isolatedfrom rice genome. These promoters were linked to the GUS reporter gene, transformed into rice,and their activity was analyzed in transgenic plants at the stages of callus and young seedling (fortress-inducible promoters), flower, leaf and roots (for tissue specific promoters). So far, we haveisolated three drought-inducible promoters, one callus-specific promoter and one root-specificpromoter. The functional assay of these promoters is ongoing. Together, we have identified andanalyzed several stress-inducible or tissue-specific promoters which would be helpful for bio-safevector construction and crops transformation. 57
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 5: Molecular breeding for cotton, brassica and bio-energy cropsProgress toward genome sequencing of upland cotton Gossypium hirsutumYu SXCotton Research Institute, CAAS, Beijing 100081, ChinaGenome sequence analysis of a plant species provides detailed resources for genomics research,including structural, functional, and evolutionary genomic studies, significantly expanding themolecular foundation for improvement of its agronomic and biological Traits. To betterunderstand the relevance of genome structure, genome size variation, and polyploidization tocotton fitness and evolution, we screened 362 BACs,about 27.5Mb, from the libraries ofGossypium hirsutum L. based on DNA markers genetically mapped on chromosomes 12 and 26.We sequenced these clones and assembled the DNA sequences into contigs. We identified fulllength LTRs, analyzed their phylogenetic relationship and found two times amplification of themajor types of LTR, one accrued before allopolyploidization, the other after theallopolyploidization. We investigated the colinearity conservation between these cotton BACsequences and with other model plant genomes, revealing that a high level component of TEscaused increased recombination in Upland cotton. The phylogenetic tree of single copyorthologues genes among cotton, Arabidopsis, Poplar, grape, rice and maize shows that poplarwas a nearer relative to cotton than others. 58
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 5: Molecular breeding for cotton, brassica and bio-energy cropsMaternal effects and genetic improvement of seed oil content in Brassica napusWang HZ*, Liu GH, Hua W, Wang XF, Liu J, Zhan GMOil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan 430062, P.R. China.* Email: wanghz@oilcrops.cnSeed oil content is an important agronomical trait in rapeseed. Our genetic crossing results haveproposed that maternal genotypes contribute significantly to the seed oil content in Brassicanapus. And silique wall as a maternal organ was found to play more important role than leaf. Inour study, using one high and one low oil content rapeseed lines zy036 (oil content 50%) and51070 (oil content 36%), we explained the mechanisms how the maternal organs act on theregulation of seed oil content.By three alternative methods, 1) F2/F3 individual correlation analysis, 2) altering the localphotosynthesis in the silique wall and 3) transcriptome and comparative genomic analysis, wefound the photosynthetic activity in silique wall was highly associated with the filial seed oilcontents. Further, OC1 and WRI, two known oil synthesis related genes in seeds, were suggestedto be involved in the regulation pathway of silique wall photosynthesis affecting oil synthesis.Additionally, one related functional gene (GK) was isolated from zy036 and the transgenic resultsin Arabidopsis implied the feasibility of enhancing the seed oil content by increasing the siliquewall photosynthetic capacity in rapeseed through genetic engineering. At the same time, thecarbon transport ability from silique wall to seed was proved to be highly related with seed oilcontent by the physiology and biochemistry studies. In this study, we also found the maternalplant heat-resistant gives a stable condition for oil synthesis in seed, and one regulated gene(hsp17) in this pathway was also isolated and proved. In sum, three pathways in the maternalorgans were considered to be involved in the regulation of seed oil content.The knowledge acquired in the study would pave the way for high oil content rapeseed breeding.Now, the highest oil content winter rapeseed cultivar (Zhongshuang 11, 49%) was generated in thenational regional test, and 5 new rapeseed lines with oil content above 55% were already bred byour genetic breeding group. 59
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 5: Molecular breeding for cotton, brassica and bio-energy cropsTowards establishing a molecular breeding platform in Cotton: Progress andChallengesKumpatla SPDepartment of Trait Genetics & Technologies, Dow AgroSciences LLC 9330 Zionsville Road, Indianapolis, IN,USACotton is the worlds leading fiber crop and the second most valuable oilseed crop. Despite itseconomic importance, the pace of genetic mapping and genomics in cotton has been slowercompared to other important crop species. This is due to the lack of development of molecular andgenetic resources such as large number of molecular markers, high density genetic linkage mapsand comprehensive physical maps as well as the non-availability of whole genome sequence ofcultivated tetraploid cotton genome. The polyploid nature, complexity of the genome and lowlevel of intra-specific polymorphism hindered the much needed establishment of marker assistedbreeding in cotton. Global efforts since late nineties, though slow, gradually resulted in thedevelopment of thousands of simple sequence repeat (SSR) markers using multiple molecular andcomputational approaches and generation of several linkage maps. However, the number ofmarkers that could be used for high fidelity molecular breeding is still very low leading severalresearchers to initiate efforts in the discovery of single nucleotide polymorphism (SNP) markers.As predicted, SNPs are abundant in cotton genome but identification of SNPs with simpleinheritance in the tetraploid like cotton poses another challenge for researchers. Several efforts arecurrently underway for large scale discovery of SNPs using high throughput sequencing andbioinformatics methods as well as to identify informative SNPs. The non availability of tetraploidcotton genome sequence poses a major challenge to these efforts. Progress in marker and mapdevelopment and challenges in establishing a marker assisted breeding platform for routine use incotton will be discussed. 60
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 5: Molecular breeding for cotton, brassica and bio-energy cropsCytological studies of apomixis in Hickory (Carya cathayensis Sarg.)Huang JQ*, Zhang B, Wang ZJ, Huang YJ, Xia GHZhejiang Agriculture and Forestry University, A Nurturing Station for the State Key Laboratory of SubtropicalSilviculture,Linan, Zhejiang Province 311300 People’s Republic of China* Email: Huangjq@zjfc.edu.cnHickory (Carya cathayensis), which is an important oil tree with high economic value whichbears edible nut in China show the tendency of apomictic embryos develop as its polyembryonyphenomemon and previous research. Up to now, it has not been reported the origin of matureembryos and its mechanism of reproduction. Hickory was identified here as a new apomicticspecies cytologically by paraffin section and flow cytometry detection. Developmental pattern ofembryo sac formation in hickory represented typical polygonum-type and female gametophytedeveloped from functional megaspore which was selected from tetrad after meiosis ofmegasporocyte. Zygote embryos had not been found in numerous histological researches andembryos all originated from nucellar cells. Initial cells of nucellar embryo located both at themicropylar end and chalazal end of embryo sac and presented with cell enlargement andvacuolization inhomogenously. Mature nucellar embryo only developed at the nucellus beakregion although bud like nucellar embryo also distributed at the chalazal end of embryo sac insome ovules absence of endosperm development which was abortion later. Ploidy of Endospermin hickory was identical with leaves of mother tree and was diploid by flow cytometry detection.Development of endosperm in hickory was spontaneous type. 61
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 5: Molecular breeding for cotton, brassica and bio-energy cropsMining of Novel Genes for Cotton Fiber Improvement 1 1 1 2Yu JZ *, Percy RG , Kohel RJ , Abdurakhmonov IY1 USDA-ARS Southern Plains Agricultural Research Center, Crop Germplasm Research Unit, College Station,Texas, USA;2 Institute of Genetics and Plant Experimental Biology, Uzbek Academy of Sciences, Tashkent, Uzbekistan.*Email john.yu@ars.usda.govMolecular breeding for cotton lags behind that for other major crops. Low levels of DNApolymorphism and genetic bottlenecks through cotton domestication complicate the efforts toimprove the genetic gains in such traits as fiber quality and yield potential. While the cottonresearch community continues to develop essential tools including polymorphic markers andgenomic sequences, we begin to take advantage of natural genetic variation that exists in largelyuntapped cotton germplasm collections using a set of core DNA markers. Association analysisof the first thousand unrelated cotton accessions, representing dozens of national origins andseveral ecotypes or landraces, with a number of fiber traits identifies unique genotypes anddiagnostic markers for cotton molecular breeding programs. Effective mining and genome-wideselection for beneficial genes or genomic fragments conferring cotton fiber and other traitsbecome possible and this approach will expand with additional molecular tools developed by thecotton research community. 62
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 5: Molecular breeding for cotton, brassica and bio-energy cropsRational design and molecular breeding of sorghum, a dedicated bioenergycropHuang YHUSDA-ARS Plant Science Research Laboratory, 1301 N. Western Road, Stillwater, OK 74075and Department of Botany, Oklahoma State University, Stillwater, Oklahoma 74078, USAEmail: yinghua.huang@ars.usda.gov Sorghum (Sorghum bicolor L.) is a C4 grass species grown worldwide because itshigh-efficiency in photosynthesis, excellent tolerance to drought and prosperity on marginal lands.This plant is a dedicated biofuel crop and has already become a viable feedstock for bioethanol. Itoffers several options in ethanol production including from its grain, sugar in sweet sorghum stalk,and vegetative biomass of the entire plant. Sorghum grain currently is the No. 2 (behind corn)source of bioethanol in the U.S. Sweet sorghum variety, similar to sugarcane, has recentlyemerged as a renewable feedstock for simple ethanol conversion. To date, cellulosic biomass isbecoming an attractive energy feedstock as the supplies are more abundant, for which sorghumshows great potential due to its unique characteristics, such as high water-use efficiency, highbiomass productivity under sustainable low-input conditions, and amenable to manipulation of itscell-wall characteristics. Sorghum has been bred for biofuel production only recently. But the recent advances ingenomics research and plant biotechnology provide opportunities to facilitate the development ofnew sorghum cultivars and hybrids for high productivity of biomass with desirable quality ofcellulose. This paper reviews the significant advances in sorghum genomics studies and the recentbreakthroughs in sorghum plant biotechnology research, and demonstrates a the potential of thegenomic and biotechnological tools for both enhancing biomass productivity and manipulatingcell wall structure and composition, which can help improve the energy efficiency of bioenergycrops through the combination of rational design and molecular breeding approach. In thispresentation, the author will highlight crop-breeding options to address biofuel needs in the futureand will also discuss opportunities and challenges in developing biomass crops for the secondgeneration of biofuels. Furthermore, the fundamental knowledge learned from the sorghum as amodel bioenergy crop can be readily applied to other fuelstock candidates such as Miscanthus andswitchgrass for their genetic improvement. 63
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 5: Molecular breeding for cotton, brassica and bio-energy cropsMolecular breeding for cottonseed quality improvementZhu SJDepartment of agronomy, Zhejiang University, Hangzhou 310029.Email: shjzhu@zju.edu.cnCottonseed, about 2/3 of the seed cotton yield, is a very important by-product in cotton production.The world seed cotton production is about 28-30 millions tons annually, which will produce about20 millions tons of cottonseed. The cottonseed, contented about 27-45% of protein and 28-40% ofoil, is a huge natural resource in plant protein and edible oil, although it has not been utilizationcompletely due to the existing of gossypol in the seeds. So the improvement of cottonseed qualityis one of the important aims in cotton breeding, but a few reports have been published up to now.An immotalized F2 (IF2) population with 188 crosses in upland cotton was constructed by randommating among recombinant inbred lines (RIL) derived from a cross of HS46 ×MARCABUCAG8US-1-88. The map genomic regions associated with seed index (SI), kernelindex (KI), kernel percentage (KP), ratio of kernel and hull (K/H), protein content (PC), oilprotein (OC), and gossypol content (GC) was identified to facilitate the selection of these traits incottonseed breeding. A linkage map consisting of 388 molecular markers from this population wasused to identify QTL using QTLNetwork-2.0 software. Seventeen main effects and fifteenepistasis QTLs were identified for these traits. Phenotypic variation explained by each individualQTL ranged from 2.42 to 23.04%. Epistatic QTL for these traits were detected and important aswell. In addition, the difference between MAS and direct selection by NIR analysis was comparedto evaluate effect of these QTLs in the breeding program targeting development of cotton withimproved nutrient quality. The efficiency of MAS for oil content was 56.4~92.5% and one SSRmarket located in chromosome 15 was linkage closely with the oil content, which can be used inMAS of cotton seed oil content. Using this marker in MAS, the oil content of more than 1000upland cotton germplasms have been selected, and 19 of them were the trait of high oil contentwhich can be used in cottonseed improvement for oil content. 64
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 5: Molecular breeding for cotton, brassica and bio-energy cropsMolecular focus in commercial plant breedingRossouw JDAsia Corn Breeding Lead, Monsanto Singapore Co (Pty) Ltd, 151 Lorong Chuan, #06-08 New Tech Park,Singapore, 556741; Dr. R.G Cantrell, Cotton Discovery Breeding Lead, Monsanto, St. Louis, MO 63167, USA.Email: jdross@monsanto.comPlant breeders have various breeding ‘tools’ to their advantage but the challenge is how toincorporate these successfully into a commercial plant breeding program to deliver products totheir farmer customers. Commercial plant breeding programs are equipment with excellentscientist and research staff, global germplasm and new breeding technologies like molecularmarkers to drive genetic gains.Commercial plant breeding has evolved over the last few decades to where we are today bycontinuously adding new technologies. During 1970-1990 the focus were on quantitativegenetics, statistics and experimental designs and having winter nurseries to have two to threebreeding cycles per year to speed the breeding process. The followed computer technologies,testing mechanization, analytics, transgenic traits, dihaploids and molecular breeding became veryimportant to drive breeding and genetic gains. Molecular breeding has yielded some greatsuccess in commercial plant breeding with successfully combining it with other aspects of plantbreeding like people, germplasm and other technologies that have been around the last fewdecades.Monsanto has a strong history of technology innovation in cotton and corn demonstrated by thegrowth in global acreage planted to Monsanto germplasm and biotech traits for insect and weedcontrol. As core crops within Monsanto; biotechnology, germplasm, and molecular breeding arebeing integrated to develop new commercial products to drive simultaneous genetic gain for lintyield and fiber quality for cotton and yield and disease resistance in corn.Looking forward GWS (Genome Wide Selection) and Whole Genome Sequencing will be thenext step in molecular technologies. GWS also opens the door on more innovation that needs tohappen across multiple disciplines to support plant breeding in the future. 65
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 6: Maize molecular breedingQTL fine mapping of leaf angle and leaf orientation value in maizeChen YHHenan Agricultural UnviersityEmail: chy9890@163.comA major limiting factor for high productivity of maize in dense planting is light penetrationthrough the canopy. Plant architecture with a narrower leaf angle (LA) and an optimum leaforientation value (LOV) is desirable to increase light capture for photosynthesis and productionper unit area. However, the genetic control of the plant architecture traits remains poorlyunderstood in maize. In this study, QTL for LA were mapped using a set of 229 F2:3 familiesderived from the cross between compact and expanded inbred lines, evaluated in threeenvironments. The results showed that one key genome region controlling leaf angle wasidentified: qLA1. The QTL at nearest marker umc2226 on chromosome 1.02 accounted for 20.4%of the phenotypic variance. Based on this, using the BC3F2 population of 300 individuals from oneBC3F1-86 plant, we mapped qLA1 to a 4.9-cM interval between bnlg1803 and bnlg1484, whichexplained 35.7% of LA variation. The BC3F2:3 families from the BC3F2 population showed a verywide variation in LA, which suggested that it was qLA1 affected the variation of LA in thispopulation. Within a qLA1 interval, single/low-copy bacterial artificial chromosome sequenceswere exploited to develop 19 polymorphic markers to saturate the qLA1 region. A step by stepnarrowing-down strategy was adopted to pursue fine mapping of the qLA1 locus. Recombinantswithin the qLA1 region, screened from each backcross generation, were backcrossed to ‘Shen137’to produce the next backcross progenies. These progenies were individually genotyped andevaluated for leaf angle. Sequential fine mapping of BC3F2, BC3F3 and BC4F2 generations enabledus to progressively refine the qLA1 locus to a ~100-kb interval flanked by the markers SSR40 andSSR53. Because of qLA1 accounting for 35.7% of LA variation, therefore, the locus was capableof improving performance of plant architecture with regard to leaf angle. 66
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 6: Maize molecular breedingApplication of Molecular Techniques in Maize Haploid BreedingChang MT1*, Li JS1 and Cai Z21 National Maize Improvement Center of China, China Agricultural University, Beijing 100193, China;2 Jilin Academy of Agricultural Science, China.* Email: cshzhang2000@yahoo.com.cnHaploid breeding is a relatively direct and rapid breeding method. Using doubled haploids (DH)to select the best maize pure elite inbred is gradually replacing the conventional maize breedingmethodology. Haploids are those plants which derived from unfertilized egg cells or sperm cells.Those plants are also called haploid sporophytes. The chromosome number of a haploid plant isonly half of their sporophyte. The size of their cells, chloroplasts and plastids are relativelysmaller, and has less DNA content. Haploid plants can be identified by their morphology,chromosome number, flow cytometry, isozyme and molecular maker analysis. The first haploidseed plant was a dwarf sea island cotton and was discovered in 1920 by O. S. Atteck. The firsthaploid maize plant was discovered in 1929 by Stadler and Randolph. The frequency ofspontaneous occurred haploids is very low and it is about 1 in ten thousands. It is possible to usevarious methods to increase the frequency of haploids, including physical treatment, chemicaltreatment, genetic induction, microspore and megaspore culture, anther culture, wild crosses,unisexual propagation or apomixis, and pollen treatments. Practical applications of haploidsinclude instant fixation of genetic pure elite inbred lines, tools for population improvement, traitconversion and cytoplasmic male sterility conversion, materials for basic genetic, quantitativegenetic, trait analysis, gene location and gene mutation study, and useful materials for molecularbreeding and basic plant research studies. Combined haploid method and molecular technique,including genomewide analysis, QTL determination, and molecular marker assisted breeding; itwill be possible to speed up the success of whole breeding process. Successful applications hasbeen applied in transgenic trait conversion and forward breeding, backward breeding, molecularmarker assisted recurrent selection, genomic wide breeding prediction, and application of DHmethod and SNP whole genome aggressive breeding strategy. 67
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 6: Maize molecular breedingIdentification of gene marker sets for screening maize lines for resistance toaflatoxin contaminationLuo M 1*, Brown RL 1, Chen ZY 2, Menkir A 3, and Bhatnagar D 11 Southern Regional Research Center, United States Department of Agriculture–Agricultural Research Service,New Orleans, LA 70124;2 Department of Plant Pathology and Crop Physiology, Louisiana State University Agricultural Center, BatonRouge, LA 70803;3 International Institute of Tropical Agriculture, Ibadan, Nigeria.* E-mail: Meng.Luo@ars.usda.govAflatoxins are the secondary metabolites of Aspergillus flavus which can be highly toxigenic andcarcinogenic to humans or animals consuming contaminated food or feeds. Presently, asignificant number of countries have established or proposed regulations for controlling aflatoxinsin food and feeds; the US Food and Drug Administration (FDA) has limits of 20 ppb, totalaflatoxins, on interstate commerce of food and feed, and 0.5 ppb of aflatoxin M1 on the sale ofmilk. However, many countries, especially in the developing world, experience contamination ofdomestic-grown commodities to alarmingly greater levels.Aflatoxin contamination caused by A. flavus is a major concern in maize production prior toharvest and during storage, and also in many other crops, such as peanuts, cottonseed, tree nuts,rice etc. Severe aflatoxin contamination during maize production is often correlated to droughtstress and high temperatures. Although many maize lines have been screened for resistance to A.flavus infection and aflatoxin biosynthesis, levels of resistance are not yet adequate to preventunacceptable aflatoxin concentrations. To rapidly identify resistant genotypes and to easily followresistance traits in breeding populations, efforts have been made to develop genetic markersassociated with resistant traits. However, very few markers are available so far.To solve this bottleneck in developing genetic markers, we are developing gene markers based oncorrelating the expression of the selected genes with the aflatoxin-resistance phenotype. Previousresearch indicated that multi-genes were involved in the resistance. Our comparative proteomicswork identified kernel resistance-associated proteins (RAPs) and demonstrated their role in kernelresistance. Our microarray investigation demonstrated that RAP genes were expressed during theobserved period from 25 to 45 day after pollination, but were expressed at a higher level at latematuration phase. In this presentation, we show that gene marker sets could be used to assessaflatoxin resistant lines, but are dependent on the particular state of kernel development. Theseresults came from investigating kernels of two closely-related maize genotypes, generated by anaflatoxin-resistance breeding collaboration between the International Institute of TropicalAgriculture (IITA) and the Southern Regional Research Center (SRRC) of the USDA-ARS. 68
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 6: Maize molecular breedingMaize disease resistance gene discovery and utilization through association andlinkage mappingMahuku G *, Yan J, Magorokosho C, Makumbi D, Crossa J, and Mataka AInternational Maize and Wheat Improvement Center (CIMMYT), Km. 45 Carr. Mexico-Veracruz. Col El Batan.Texcoco-Edo Mexico. Mexico CP 56130* Email: g.mahuku@cgiar.orgAssessing the association between DNA variants and disease has been used widely to identifyregions of the genome and candidate genes that contribute to disease resistance. Identifying therelevant genes has been difficult, in part because each causal gene only makes a smallcontribution to overall heritability. Genetic association studies assess correlations between geneticvariants and trait differences on a population scale, and offer a powerful approach for identifyingand mapping causal genes with modest effects. In CIMMYT, we are using nine environmentsspread in Africa and Latin America to phenotype over 300 advanced maize lines against maizestreak virus (MSV), northern corn leaf blight (Excerohilum turcicum), southern corn leaf blight(Bipolaris maydis), common rust (Puccinia sorghi), gray leaf spot (Cercospora zeae-maydis), andfusarium ear rots (Fusarium verticillioides). The objectives of this study are to (i) identify diseaseresistance donors effective across environments and to use association mapping strategies todetect markers near to esistance genes in the maize genome and subsequent development ofgene-based markers for use to aid selection in our breeding programs. Our results showed highrepeatability (r = 0.7 to 0.99) within locations compared to between locations (r = 0.30 to 0.40).These results revealed that while some sources of resistance are good across localities, pathogenpopulations are also highly variable and might be different between locations. Therefore, potentialsources of resistance should be evaluated in multiple environments to select best genes to use in abreeding program. Marker-trait association using the general and mixed linear models identified49 SNPs that were significantly associated with disease resistance. Thirty-four of these SNPmarkers were unique. Haplotype analysis using significant SNPs revealed that additive geneaction appears to be more important in disease resistance than epistasis. The use of thisinformation to elucidate the genetics of disease resistance in maize, organization of diseaseresistance genes in the maize genome and prospects for development of gene-based markers arediscussed. 69
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 6: Maize molecular breedingForward to Molecular Breeding for High-oil MaizeYang XH 1, Hao XM 1, Ma HL 1, Gou YQ 1, Yan JB 1, 2, Song TM 1, Li JS 1*1 National Maize Improvement Center of China, China Agricultural University, Beijing 100193, China;2 International Maize and Wheat Improvement Center (CIMMYT), Apdo. Postal 6-641, 06600 Mexico, D.F,Mexico* Email lijiansheng@cau.edu.cnMaize oil is a highly valued crop as animal feed, human food and biodiesel. Phenotypic variationof oil content in maize kernel could be related to oil components like fatty acids and physicalcharacteristics of the kernel as embryo size and embryo to endosperm weight ratio. To determinethe genetic basis of oil content and composition in maize grain, a recombinant inbred populationderived from a cross between normal line B73 and high-oil line By804 was phenotyped using gaschromatography, and was genotyped with 228 molecular markers. A total of 42 individual QTL,associated with fatty acid compositions and oil content, were detected in 21 genomic regions. Fivemajor QTL were identified for measured traits, one each of which explained 42.0% of phenotypicvariance for palmitic acid, 15.0% for stearic acid, 27.7% for oleic acid, 48.3% for linoleic acid,and 15.7% for oil content respectively in the RIL population. Thirty-six loci were involved in 24molecular marker pairs of epistatic interactions across all traits, which explained phenotypicvariances ranging from 0.4 to 6.1%. These results demonstrated that a few major QTL with largeadditive effects could play an important role in attending fatty acid compositions and increasingoil content in used germplasm. A larger number of minor QTL and a certain number of epistaticQTL, both with additive effects, also contributed to fatty acid compositions and oil content.In order to further identify the causal variants for QTL controlling oil content in maize kernel,seven more oil related traits (embryo oil content, embryo oil density, embryo to endosperm weightratio, embryo volume, embryo width, and embryo length and embryo width to length ratio) weremeasured in the same RIL population. A total of 58 QTLs were identified for oil content in maizekernel and its component traits in 26 genomic regions across all chromosomes. Eight main-effectQTLs were identified for kernel oil content, embryo oil content, embryo oil concentration, embryoto endosperm weight ratio, embryo weight and embryo width to length ratio, each accounting forover 10% of the phenotypic variation in 6 genomic regions. Over 90 % of QTLs identified forkernel oil content co-localized with QTLs for component traits, validating their molecularcontribution to kernel oil content. As kernel oil content is dependent on many factors, identifyingQTL associated with component traits of oil content provided additional information on thegenetic basis of kernel oil QTL. On chromosome 1, the largest effect QTL for kernel oil content(qKO1-1) was associated with embryo width, while on chromosome 9, the QTL for kernel oilcontent (qKO9) was related to embryo to endosperm weight ratio. Embryo oil density and embryo 70
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京width were identified as the most important component traits controlling the second largest QTLfor kernel oil content on chromosome 6 (qKO6) and a minor QTL for kernel oil content onchromosome 5 (qKO5-2), respectively. The dissection of kernel oil QTLs will facilitate futurecloning and/or functional validation of kernel oil content, and help to elucidate the genetic basis ofoil content in maize kernel.Based on above results, two major QTL regions, qKO1-1 in chromosome bin 1.04 and qKO6 inchromosome bin 6.04, were targeted for introgression of the favorable alleles from the high-oilline into two elite inbred lines through molecular maker assistant selection. Chang7-2 andZheng58, which are the parents of widely extended hybrid Zhengdan 958 in China, were used forrecurrent parents. The BC5F2:3 lines were evaluated for oil content and background recovery. ForqKO1-1, the introgressed genomic fragment was about 20cM containing the targeted QTL, andthe oil content of maize ears with high-oil homologous genotype was increased by 0.6 and 0.3%for the improved lines from chang7-2 and zheng58, respectively. For qKO6, a function markerwas developed from the validated gene DGAT1-2 controlling qKO6 and used in maker-assistedselection. In BC5F2:3, the introgressed genomic fragment contained the entire gene DGAT1-2.The oil content of maize ears with high-oil homologous genotype was increased by 0.8 and 0.5%for the improved lines from chang7-2 and zheng58, respectively. These results show MAS is apromising way to improve the quantity traits using QTL with large effects in maize, especially forthe cloned QTL with validated function and developed function markers. 71
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 6: Maize molecular breedingGenome-wide association study identifies known as well as novel loci for maizekernel tocopherol content and compositionLi Q 1*, Xu ST 1, Yang XH 1, Gao SB 2, Zhang ZX 3, Cai Y 1, Zhang DL 1, Zhou Y 1, Li JS 1, *, Yan JB 1, 4, *1 National Maize Improvement Center of China, China Agricultural University, 100193 Beijing, China.2 Maize Research Institute, Sichuan Agricultural University, 625014 Yaan, Sichuan, China.3 National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, 430070 Wuhan, Hubei,China.4 International Maize and Wheat Improvement Center (CIMMYT), Apdo. Postal 6-641, 06600 Mexico, D.F., Mexico* Email: lijiansheng@cau.edu.cn; yjianbing@gmail.comTocopherols are important antioxidants for both human health and plant development. Here, wetry to identify genetic variants that can control maize kernel tocopherol content and compositionusing both genome-wide association study (GWAS) and candidate gene based association study.Three kinds of tocopherols, namely δ-, γ- and α-, were scored in a panel of 513 lines across threelocations. The contents ranged from 0.55 to 8.93 µg/g for δ-tocopherol, 3.35 to 141.07 µg/g forγ-tocopherol, 0.38 to 60.92 µg/g for α-tocopherol, 7.46 to 198.35 µg/g for total tocopherol;respectively. And the ratio of α-tocopherol to γ-tocopherol varies between 0.02 and 3.49. GWASwith about 50,000 SNPs identified 20 variants that are associated with at least one of the five traits(P < 1.02 × 10-6). The strongest association was on chromosome 5 for α-tocopherol, where acandidate gene from tocopherol biosynthesis pathway was located. Re-sequencing of this gene ina subset of 155 lines identified a total of 206 polymorphisms, most of which are in significantlinkage disequilibrium. An insertion/deletion polymorphism (InDel) was significantly associatedwith α-tocopherol and also segregated in a recombinant inbred line population where aquantitative trait locus for the same trait was identified. This InDel can increase/decreaseα-tocopherol content by 11.27-25.48 µg/g in four F2 populations with different geneticbackground. Other significant variants are from genes with no obvious function in the tocopherolbiosynthesis pathway, including transcriptional factor, genes targeted to chloroplast ormitochondria and so on. Re-sequencing of other five candidate genes from tocopherolbiosynthesis pathway didn’t identify any strong associations, indicating these genes might playlimited role in tocopherol phenotypic diversity. 72
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 7: Applied plant genomics: from genomics to fieldMolecular breeding in chickpea- still a dream or the reality now!RK Varshney1,2,*1 International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Patancheru-502324, India;2 Theme- Comparative and Applied Genomics, Generation Challenge Programme (GCP), c/o CIMMYT, IntAPDO Postal 6-641, 06600 Mexico DF, Mexico* Email: r.k.varshney@cgiar.orgChickpea (Cicer arietinum L.) is the most important cool season food legume crop, cultivated inarid and semi-arid region of the world. Terminal drought and Helicoverpa are two majorconstraints to chickpea production. Until a few years ago, implementation of molecular breedingprogramme was not possible in chickpea, due to dearth of sufficient genomic resources. With anobjective to enhance molecular breeding in chickpea, concerted efforts of ICRISAT incollaboration with several partners around the world, lead to development of large scale genomicresources (http://www.icrisat.org/gt-bt/ICGGC/homepage.htm). For instance, 1,655 novel SSRshave been isolated from the SSR-enriched library (311) and BAC-end sequences (1,344), a DArTarray has been developed with >16,000 features and >20,000 ESTs from drought and salinitystress challenged tissues based on Sanger sequencing. Further, a set of 443,969 sequence tagswere generated through FLX-454 sequencing from a pool of normalized cDNAs assembled fromdevelopmental stages and abiotic stresses challenged tissues of a reference chickpea genotype(ICC 4958). Analysis of Sanger as well as FLX-454 sequence data provided 103,215 tentativeunique sequences (TUSs). Alignment of >118.2 million sequence tags generated from droughtresponsive (ICC 4958 and ICC 1882) and Helicoverpa challenged tissues (ICCC 37 and ICC 506)with TUSs provided >90,000 nucleotide variants (SNPs) in these genotypes. In collaboration withUniversity of California-Davis, a pilot Illumina GoldenGate assay for 768 SNPs coming fromconserved genes across legumes has been developed. By using above mentioned resources, anintegrated genetic map with >1400 marker loci has been developed based on the inter-specificmapping population (C. arietinum ICC 4958 × C. reticulatum PI 489777). With an objective ofidentification of candidate markers associated with drought tolerance (root traits), twointra-specific mapping populations (ICC 4958 × ICC 1882 and ICC 283 × ICC 8261) have beenphenotyped for root traits in two environments and genotyped with >300 SSR markers.Marker-trait analysis has revealed several QTLs including one major QTL in both mappingpopulations that contributed up to 36% phenotypic variation. This genomic region is beingintrogressed into three elite chickpea lines (JG 11, ICC 92318 and KAK 2), using marker-assistedbackcrossing (MABC) approach, to develop superior cultivars with enhanced drought tolerance.Due to availability of these genomic resources, molecular breeding in chickpea comes of age andis expected to be the integral part of chickpea breeding in near future. 73
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 7: Applied plant genomics: from genomics to fieldSingle-base resolution DNA methylomes of rice and new regulatory roles ofDNA methylation in plant gene expressionLi XKunming Institute of Zoology, Chinese Acadmey of Sciences, No.32 Jiaochang DongluKunming 650223, Yunnan, ChinaEmail: lixin@mail.kiz.ac.cnDNA methylation plays important functional roles in plants and animals. To examine rice genomemethylation landscape and assess its functional significance, we generated the first single-baseresolution genome methylation maps for Asian cultivated rice Oryza sativa ssp. japonica, indicaand their wild relatives, Oryza rufipogon and Oryza nivara. The overall methylation level of ricegenomes is four times higher than that of Arabidopsis. Methylation in promoters represses geneexpression while gene-body methylation generally appears to promote gene expression, which isconsistent with the results reported for Arabidopsis but different from the previous epigeneticstudies on rice. Most interestingly, we discovered that methylation in gene transcriptionaltermination regions can significantly repress gene expression, and the effect is even stronger thanthat of promoter methylation, which suggests a new direction in the study of DNA methylation.Through integrated analysis of genomic, DNA methylomic and transcriptomic differencesbetween cultivated and wild rice, we found that the genetic factor reflected by DNA sequencedivergence may be the major determinant for methylation differences at the whole genome level,but DNA methylation difference can only account for limited gene expressional variation betweencultivated and wild rice. We also identified a number of genes with significant methylation leveldifference between wild and cultivated rice. 74
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 7: Applied plant genomics: from genomics to fieldInsertion Site-Based Polymorphism markers open new perspectives for genomesaturation and marker-assisted selection in wheatCubizolles N1, Laugier C1, Mastrangelo AM2, Faure S1, Choulet F1, Roger D3, Gauthier V3, Martinant JP3,Sourdille P1, Balfourier F1, Le Paslier MC4, Chauveau A4, Cakir M5, Gandon B3, Guerreiro L6, Gielen J7,Jaubertie JP8, Jack P9, Robert O10, Korzun V11, Feuillet C1 and Paux E1*1 INRA UBP UMR1095, 63100 Clermont-Ferrand, France; 2Centro di Ricerca per la Cerealicoltura, 71122Foggia, Italy; 3Limagrain Verneuil Holding, 63204 Riom, France; 4INRA-EPGV UR1279, 91057 Evry, France;5 Murdoch University, Murdoch WA 6150, Australia; 6Arvalis-Institut du Végétal, 75116 Paris, France ;7 Syngenta Seeds, 31790 Saint Sauveur, France; 8AgriObtentions, 78041 Guyancourt, France ; 9RAGT Seeds,Ickleton, Essex CB10 1TA, UK; 10Bioplante, 59930 La Chapelle dArmentières, France ; 11KWS LOCHOW,37574 Einbeck, Germany.*Email: etienne.paux@clermont.inra.frIn wheat, the deployment of marker-assisted selection has long been hampered by the lack ofmarkers compatible with high-throughput cost-effective genotyping techniques. Recently,Insertion Site-Based Polymorphism (ISBP) markers have appeared as very powerful new tools forgenomics and genetic studies in hexaploid wheat. To demonstrate their possible use in wheatbreeding programmes, we assessed their potential to meet the five main requirements forutilization in MAS: flexible and high-throughput detection methods, low quantity and quality ofDNA required, low cost per assay, and tight link to target loci and high level of polymorphism inbreeding material. Toward this aim, we developed a programme, IsbpFinder, for the automateddesign of ISBP markers and adapted three detection methods (melting curve analysis, SNaPshot®Multiplex System and Illumina BeadArray technology) for high throughput and flexible detectionof ISBP or ISBP-derived SNP markers. We demonstrate that the high level of polymorphism ofthe ISBPs combined with cost-effective genotyping methods can be used to efficiently saturategenetic maps, discriminate between elite cultivars, and design tightly linked diagnostic markersfor virtually all target loci in the wheat genome. All together, our results suggest that ISBPmarkers have the potential to lead to a breakthrough in wheat marker-assisted selection. 75
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 7: Applied plant genomics: from genomics to fieldIntegrating technologies for genetic improvement of quantitative traits insorghumMace E*, Sakrewski K, Hunt C, Shatte T, Cruickshank A, Henzella R, Jordan DDepartment of Employment, Economic Development and Innovation (DEEDI), Queensland, Australia;* Email: emma.mace@deedi.qld.gov.auOvercoming abiotic stresses and improving yield in crops remains one of greatest challenges inplant breeding. Understanding the genetic factors, and their interactions with the environment,that underlie such quantitative phenotypes is complex and requires integration across technologies,including high throughput genotyping and sequencing. Sorghum, a C4 cereal, is in a uniqueposition to make use of a range of technologies with its rich history of genome analysis, fromgenetic linkage and physical mapping, through to the recent completion and annotation of thewhole genome sequence, providing opportunities to fast-track progress towards betterunderstanding gene function in cereals. A set of coordinated germplasm and technology resourcesis described which have been developed within the sorghum breeding program (DEEDI) toenhance the genetic dissection and improvement of both qualitative and quantitative traits insorghum. These resources link to the whole genome sequence and include a Nested AssociationMapping (NAM) population associated with phenotypic data across multiple environments and amutagenised population consisting of approximately 5000 individuals, and encompass bothforward and reverse genetics approaches to gene function determination in sorghum, representinga unique resource for the sorghum community to deliver advances in efficiency in research anddevelopment programs focused on complex traits. We detail a study focusing on the determinationof the impact of allelic variation of key flowering time genes, and their interaction with theenvironment, detailing the combined use of the NAM populations, structured mappingpopulations and reverse genetics resources. 76
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 7: Applied plant genomics: from genomics to fieldIrradiation mutant mapping of wild beet translocation lines carrying resistancegenes against the beet cyst nematodeCapistrano G *, Jäger SC, Harloff H, Jung CPlant Breeding Institute, Olshausenstr. 40, 24098 Kiel, Germany*e-mail: g.capistrano@plantbreeding.uni-kiel.deThe beet cyst nematode Heterodera schachtii Schmidt (BCN) is a severe pest in sugar beet (Betavulgaris L.) and the only sources of complete resistance are the wild species Patellifoliaprocumbens and the related species P. patellaris. Sugar beet translocation lines A906001 andTR363 carrying a translocation of P. procumbens chromosome 1 are fully resistant to BCNs. Thenematode resistance gene Hs1pro-1 had been cloned from the translocation line A906001 using aYAC based 1st generation physical map. However TR363 does not carry this gene. Hs1pro-1 gavecomplete resistance in a complementation study done with sugar beet hairy roots but only partialresistance was found in whole sugar beet plants transformed using this gene. These were strongindications for a second resistance gene on this translocation. Because the whole translocation isexcluded from recombination, chromosome breakage mutants were produced. Seeds were 400 Gygamma irradiated to produce breakages within the translocation and thus narrow down to theresistance gene. The mutants were screened with translocation specific markers for theidentification of lines with smaller translocations. 2,670 seeds were irradiated and, after screeningof 578 M1 offspring with three molecular markers evenly spread around the translocation, twomutants were found. These mutants, named TR320 and TR659, are susceptible to the BCN and hadlost most of the translocation region. A BAC based physical map was established with a minimaltiling path encompassing 18 BACs covering 1,484 kb of the translocation region which had beenestimated to be 1,500 kb in size. The BAC contigs contain only two small gaps. By colinearityanalysis between four wild beet translocation lines, the region housing the nematode resistancegene could be narrowed down to ca. 180 kb. A candidate gene residing within this critical region isfunctionally characterized by transformation into sugar beet hairy roots and A. thaliana. 77
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 8: Rice molecular breedingDevelopment of 384-plex SNP marker sets for diversity analysis, mapping, andmarker-assisted selection in riceThomson MJ*1, Zhao K2, Wright M2, McNally KL1, Leung H1, McCouch SR21 International Rice Research Institute, DAPO Box 7777, Metro Manila, Philippines;2 Cornell University, Ithaca, NY 14853, USA*Email: m.thomson@cgiar.orgMarker-assisted selection can enable more precise and rapid breeding strategies, but limitations incurrent genotyping techniques has prevented markers from being integrated into manymainstream breeding programs. Multiplexed single nucleotide polymorphism (SNP) markershave the potential to increase the speed, efficiency and cost-effectiveness of marker genotyping,provided that an optimal SNP density is used for each application. To test the usefulness ofmultiplexed SNP genotyping in cultivated rice, we designed four GoldenGate oligo pool assay(OPA) sets using Veracode technology for the Illumina BeadXpress Reader. Validated markersfrom existing 1,536 Illumina SNPs and 44K Affymetrix SNP chips developed at CornellUniversity were used to select subsets of SNPs for maximum information and even distributionfor each application. A 96-plex OPA was developed for assigning a sample into one of the fiveOryza sativa population subgroups in rice: indica, tropical japonica, temperate japonica, aus andaromatic. One 384-plex OPA was designed to have evenly-spaced polymorphic markers forQTL mapping, background selection and diversity analysis for indica and aus germplasm, whiletwo additional 384-plex OPAs were selected for use in indica/japonica crosses and for DNAfingerprinting. The cost-effectiveness of each SNP set for different mapping populations willdepend on the percent polymorphism across different combinations of mapping parents. Afterinitial testing and validation on diverse germplasm accessions and populations, two of the SNPsets were re-designed to replace poor-performing markers. Further research is required toidentify relevant SNPs for additional germplasm pools such as wild Oryza species. Theavailability of optimized SNP sets will help increase the efficiency of DNA fingerprinting, QTLmapping and marker-assisted selection to enable more rapid variety development to meet thechallenges of rice improvement in the future. 78
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 8: Rice molecular breedingEpigenetic and Genetic Control of Drought Tolerance in Rice – A mergingstory of Larmarkism and MendelismLi ZK *,1,2, Dwivedi DK2,3, Lafitte R2, Liu SH1,2, Gao YM1,2, Xu JL1,2, Zhang F1, Fu BY1,2, Zheng TQ1,2, Ali J2,Wang Y1, Cui YR1, Domingo JR2, and Venus E21 Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China; 2 InternationalRice Research Institute, DAPO Box 7777, Metro Manila, Philippines; 3 Department of Biotechnology, N. D.University of Agriculture and Technology, Narendra Nagar (Kumarganj) 224 229, Faizabad (U.P.), India.*Email: z.li@cgiar.orgDrought is the most important factor limiting rice yields in the rainfed areas of Asia and Africa. Toovercome this problem, major efforts have been taken to develop drought tolerant (DT) ricevarieties and understand the genetic basis of (DT) in rice. Three sets of experiments wereperformed. First, we developed 221 IR64 introgression lines (ILs) with significantly improved DTselected from 13 BC2F2 populations from crosses between IR64 (recipient) and 7 diverse donors.Genetic characterization using SSR markers revealed an average of over-representation of thedonor homozygotes by 1.7 times and a genomewide reduction of heterozygosy by 68% in the ILs.The second set of experiments included genotypic and phenotypic characterization of 455 DT F3pyramiding lines (PLs) selected under severe drought from 9 F2 populations derived from crossesbetween 15 DT IR64 ILs. Genetic analyses with SSR markers revealed a genomewide epigeneticsegregation (ES) in the DT PLs characterized with complete loss of heterozygosity at mostsegregating loci, strong non-random associations between or among unlinked loci of ES, and thepresence of cryptic (inversions and translocations) variation in virtually all genomic regions of ES.Comparisons in segregation patterns between the drought selected PLs and random F2 progeny ofthe same 9 populations revealed two types of epiloci, ~30% of which inherited ES from theirparental ILs and the remaining 70% were drought induced. LD analyses revealed strongnonrandom associations among the favorable alleles at these epiloci, forming putative geneticnetworks underlying DT in rice. The PLs from each population consisted of only a few majorgroup (multi-locus) genotypes (GGs), which showed significantly improved DT over IR64, butdiffered considerably in yield and some other traits under irrigated conditions. The third set ofexperiments included 667 2nd round PLs selected under severe drought from the F2 populationsof 14 crosses between 8 1st round PLs. Surprisingly, the drought selected F2 progeny of thesepopulations restored completely or partially to the Mendelian segregation. Based on the results,we proposed a hypothesis and putative mechanism(s) based on which how and under whatconditions Mendelian and ES (Larmarkian) segregation could be converted to each other. Finally,we proposed an efficient molecular breeding strategy for genetic improvement of DT and otherabiotic stress tolerances in plants by designed QTL pyramiding and taking advantage of epigeneticcontrol of these traits. 79
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 8: Rice molecular breedingClustered QTLs for source leaf size and yield traits in riceWang P1, Zhou GL1, Cui KH1, Li ZK1,2, Yu SB1*1 National Key Laboratory of Crop Genetic Improvement, and The College of Plant Science and Technology,Huazhong Agricultural University, Wuhan 430070, China.2 National Key Facility for Crop Gene Resources & Genetic Improvement, Chinese Academy of AgriculturalSciences, Beijing 100081, China*Email: yusb@mail.hzau.edu.cnImprovement of plant type plays an important role for super-high yield breeding in rice. In thepresent study, a set of backcross recombinant inbred lines derived from the cross of ‘9311’ and‘Zhenshan97’, both elite indica hybrid rice parents, were developed to identify quantitative traitloci (QTL) for flag leaf size, panicle and and yield traits. Forty-seven QTLs were detected incommon for 14 traits in the two environmental trials, of which 9 genomic regions with clusteredQTLs affecting plant type and yield traits. Four co-localized QTLs on chromosomes 1, 6, 7 and 8involving QTLs for flag leaf size (flag leaf length, width and area) contained the QTLs for yieldtraits such as panicle weight (PW) and secondary branch number (SBN), and in all cases allelesfrom ‘9311’ increased source leaf size were associated with increased sink size and yield (SBNand PW). Using a subset of overlapping substitution lines for the QTL region on chromosome 1,we validated and narrowed the QTLs into a 990 kbp interval (RM3746-RM10435) with thepleiotropic effects on flag leaf size, PW, SBN and spikelet number per panicle. These QTLclusters with large effects on source leaf size and yield-related traits provide good targets formarker assisted breeding for high-yield potential in rice. 80
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 8: Rice molecular breedingMolecular breeding approaches for sustainable disease resistance in rice:Current and future strategiesVera Cruz CM1, Jena KK1, Choi IR1, Mauleon RP1, Kobayashi N1, Fukuta Y2, Ali J1, Li Z1,3, Satoh K3, KikuchiS3, Leung H11 International Rice Research Institute, DAPO Box 7777, Metro Manila, Philippines;2 JIRCAS, Tsukuba, Japan;3 Chinese Academy of Agricultural Sciences, 12 Zhongguancun South Street, Beijing 100081, China;4 NIAS, JapanRice diseases have always been a significant factor in rice supply as new rice varieties and hybridsare planted to millions of hectares. Minor diseases and new forms of pathogens also evolve withchanges in crop production practices and climate. Under these circumstances, disease resistance isstill considered the most effective and desirable disease control method in terms of economics andenvironment safety. Thus, breeding for disease resistance has been an integral component of riceimprovement and disease management strategies. Bacterial blight, blast, brown spot and tungroresistance are among the key diseases where qualitative and quantitative resistance associatedwith major genes and candidate defense genes have been identified and proven effective inendemic situations. Some effective or broad spectrum R genes are xa5, xa13, Xa7, Xa21, Xa23 forbacterial blight, Pi2, Pi9, Pi40, Piz, Piz5, Pik, Pi1, Pi7 for blast, tsv1 for tungro resistance andthese are available for incorporation in elite parental lines for inbred and hybrid rice breeding.QTL regions containing some members of the germins and germin-like gene family have beenassociated with blast resistance. A recessive gene (tsv1) in chromosome 7 encoding initiationelongation factor (IEF) is strongly associated with RSTV resistance. Availability of gene-basedmarkers and functional SNPs from these genes will contribute to mainstream molecular breedingfor combining disease resistance with abiotic stress tolerance and grain quality traits in targetenvironments.To sustain breeding for multiple disease resistance combined with other useful traits, it is essentialto continue the identification of resistance sources from a wide array of germplasm with the aid ofhigh-throughput genotyping and phenotyping methods. The evolution of the next generationsequencing approaches and transcriptome analysis can provide a genome-wide view of genes andQTL regions for disease resistance that would enable selection and combination of desirable genesand QTL regions. To improve breeding efficiency, we propose (a) the development ofbreeding-ready NILs for important major R genes and QTL, (b) greater exchange of breedingmaterials between countries for evaluation at disease hotspots in target countries, and (c) renewedmonitoring of pathogen populations with differential varieties or functional pathogen diagnosticmarkers. Strong partnerships and enhanced linkages within countries are needed for successfulimplementation of these approaches to develop sustainable disease resistance in breedingprograms. 81
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 8: Rice molecular breedingMAS pyramiding of disease and pest resistant genes into drought toleranthybrid riceLiu GL, Mei HW*, An ZS, Li TF, Feng FJ, and Luo LJ*Shanghai Agrobiological Gene Center (SAGC), Shanghai 201106, China*Email: hmei@sagc.org.cn or lijun@sagc.org.cn)To face the challenges of food and water safety, water saving and drought resistant rice (WDR) is the priorobjective of research actitities in SAGC. A series of varieties, both inbreds and hybrids, have been developedand released in past ten years. Among them, one hybrid combination “Hanyou No3” has good performancewhen tested in several provinces in southern China, including high yield potential, good grain quality, lodgingresistance and adapting to light cultivation like direct sowing with no tillage.Two parental lines of this hybrid, “Huhan 1B” as the CMS maintainer and “Hanhui No3” as the restorer, wereused as the recipients in this study. Following the gene mapping and cloning results reported in publications,in-gene or tightly linked molecular markers with polymorphism between donor and recipient were developedfor each gene. Marker assisted backcross breeding procedure was initiated two years ago to introgress, and topyramid several resistant genes to rice blast (Pi1, Pi2 and Pi9), bacteria leaf blight (Xa21 and Xa23) and brownplant hopper (Bph14 and Bph15) into both parental lines.Using both foreground and background selections in large-size segregating populations, a number of candidatelines have been developed that carry one to three two resistant genes. Preliminary tests showed the resistancesof lines with target genes had improved resistances in comparison with the recipient parents and sister lineswithout target genes. Genes controlling phosphorus uptaking (Pup1) and fragrance (fgr) were also included inthis breeding program. 82
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 8: Rice molecular breedingDevelopment of Single Nucleotide Polymorphisms (SNPs) Detection Platformsfor Genetic Analyses and Molecular Breeding of RiceChen HD1, *, He H1, Zou YJ 1, Chen W1, Yu RB1, Yang Y1, Gao YM2, Xu JL2, Zhou SC3, Li Y1, Li ZK2, andDeng XW11 College of Life Sciences, Peking University, Beijing 100871, China.2 Chinese Academy of Agricultural Sciences, Beijing 100081, China.3 Guangdong Academy of Agricultural Sciences, Guangdong 510640, China.* E-mail: chenhaodong@gmail.comMolecular markers are used more and more in rice breeding. Single nucleotide polymorphisms(SNPs) show a lot of advantages comparing to other markers, which can be used widely inhigh-density genetic maps construction, association mapping and marker assisted selection. Here,around 132,000 SNPs meeting the criterion for our SNPs detection assays were selected bycomparing the genome sequences of several rice varieties including Minghui 63, Zhenshan 97,PA64S, 9311 and Nipponbare. 3090 SNPs distributed evenly along the rice whole genome wereselected and confirmed by PCR and sequencing, and most of them were considered to be suitablefor both Illumina GoldenGate assay and optical thin-film biosensor chips, two platforms we havegenerated. A set of 384 SNPs were further selected to analyze around 350 global rice breedingvarieties and a pedigree of elite variety Huanghuazhan. The phylogenetic tree revealed severaldistinct groups, and the pedigree analysis showed interesting conserved and changed sites alongthe rice genome during the breeding process. These platforms provide a foundation for furtherdiversity analysis, mapping and marker-assisted selection in rice research. 83
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 8: Rice molecular breedingIdentification of a new blast resistant gene from Dacca6, a useful donor toimprove the wide spectrum resistance of Jin23 against rice blast fungi(Magnaporthe grisea) in Southeast ChinaShi BH1, Zhang JH1, Zheng YM2, Liu YQ2, Zheng TQ3,*, and Zhao MF2,*1 College of Life Sciences ,Fujian Normal University, Fuzhou, Fujian 350108, China;2 Rice Research Institute, Fujian Academy of Agricultural Sciences, Fuzhou, Fujian 350018, China;3 Institute of Crop Sciences/National Key Facility for Crop Gene Resources and Genetic Improvement, ChineseAcademy of Agricultural Sciences, 12 South Zhong-Guan-Cun St., Beijing 100081, China.* Email: tzheng@caas.net.cn & zhmingfu@163.com.Jin23 is a major parental line being widely used in China’s three-line hybrid rice breeding. Itsexcellent grain quality has filled the gap between the high yield potential and good grain qualitywhich bothering the hybrid rice breeding for quite a long time. However, it was quite sensitive torice blast fungi (Magnaporthe grisea), especially in Southeast China, such as Fujian province. Inorder to improve the resistance of Jin23, new genetic resources are highly required. In 2005, 130molecular breeding parental lines imported from the global rice molecular breeding networkinitiated at IRRI have been screened by natural inoculation at the blast endemic field in Fujian.Among them, Dacca6, a local variety from the Philippines, was found to possess wide-spectrumand high resistances to the major races in Fujian. The following report focused on the furtheridentification of the resistance by artificial inoculation and the mapping of the underlying geneticlocus as well as its application in the improvement of Jin23B (maintainer) for the blast resistances.Two subsets of a BC1F2 population derived from the cross of Dacca6 with Jin23B were inoculatedby spraying with two representative isolates, SM03023 and SH0512, respectively, at the seedlingstage. Both of the segregation results from these two subsets pointed to a same new singledominant gene from Dacca6. By traditional linkage mapping, this gene was tagged betweenmarkers RM5529 and RM211, the genetic distance between these two markers and the gene was3.8 cM and 4.1 cM, respectively, on chromosome 2. Furthermore, we assayed the DNA pools ofresistant and susceptible individuals from independent BC1F2 populations derived from eightBC1F1 resistant plants with 21 SSR polymorphic makers on chromosome 2 to identify theintrogression patterns. It was found that all the pools were heterozygous at the marker locus ofRM211, which indicated that the resistance gene was closer to RM211. This was consistent withthe results from the previous linkage mapping work. With the aid of comparative mapping tools,we compared the candidate position with the known blast resistant genes, and no reported genewas found in this region. The existence of a new resistant gene in this candidate region wasstrongly indicated.Based on the mapping results, after consecutive backcrossing followed by selfing with the aid of 84
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京molecular marker aided selection (MAS) by RM5529 and RM211, twelve Jin23B BC3F3 lineswere obtained. The resistances of these lines were confirmed by a field evaluation in the blastepidemic area, Jiangle County, Fujian. One of these elite lines was further evaluated for itsagronomic traits, and no significant difference was found in comparing with Jin23B. This study has preliminarily showed a way to breeding blast resistant Jin23B with new genefrom Dacca6 by MAS. The development of blast resistant Jin23 male sterile line is undergoingbased on the lines from this work. Meanwhile, the mapping information will also help on therelated gene cloning work in the near future. 85
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 9: Wheat molecular breedingGenomic and proteomic studies of gliadin proteins in bread wheat (Triticumaestivum L.)Wang D1, Wang DW1*, Zhang X2, Yue G1, Li Y1, Qin G21 State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and DevelopmentalBiology, Chinese Academy of Sciences, Beijing 100101, China2 Key Laboratory of Ion Beam Bioengineering, Zhengzhou University, Zhengzhou 450001, China* E-mail: dwwang@genetics.ac.cnGliadins are conserved seed storage proteins in wheat and related Triticeae species. Althoughclassical genetic and biochemical investigations have shown that the genes resided in Gli-1 andGli-2 loci encode gliadins, systematic information on the genomic organization, recombination,expression and functional characteristics of gliadin genes is still lacking. We have used genomicand proteomic approaches to study the gliadin genes in two elite bread wheat varieties Xiaoyan 54and Xiaoyan 81. The BAC clones harboring different types of gliadin sequences have beenidentified, several of which are being sequenced to aid our understanding of the genomicorganization of gliadin genes in bread wheat. A panel of mutants missing the expression ofdifferent types of gliadin proteins has been identified via the use of mass spectrometry. Thesedeletion mutants are being employed in proteomic experiments to establish the relationshipsbetween gliadin genes and their products accumulated in the seeds. A backcrossing program hasalso been initiated to purify the genetic background of the deletion mutants, which will make themuseful for further studies of the effects of gliadins on the processing quality and yield potential ofbread wheat. 86
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 9: Wheat molecular breedingDevelopment and Application of Molecular Markers for Improving ProcessingQuality in Common WheatHe ZH*1,2, Xia XC11 Crop Science Institute, Chinese Academy of Agricultural Science (CAAS), Zhongguancun South Street 12,Beijing 100081, China2 CIMMYT China Office, C/O CAAS, Zhongguancun South Street 12, Beijing 100081, China* Email: zhhecaas@163.comImprovement of processing quality in common wheat is an important breeding objective in Chinaand application of molecular markers will improve the efficiency of breeding program. Functionalmarkers for high molecular weight gluten subunits and low molecular weight gluten subunitsresponsible for gluten quality, phytoene synthase 1 (Psy-1) gene and lipoxygenase (LOX) activityassociated with yellow pigment and polphenol oxidase (PPO) activity, controlling for color ofend-use products, were developed and validated in Chinese and CIMMYT wheats. Our experienceindicated that integration of conventional testing and utilization of molecular markers willimprove the efficiency of quality improvement program. 87
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 9: Wheat molecular breedingNew Insights into the Organization, Recombination, Expression andFunctional Mechanism of Low Molecular Weight Glutenin Subunit Genes atthe Complex Glu-3 Loci in Bread WheatDong LL, Zhang XF, Liu DC, Fan HJ, Sun JZ, Hao ST, Li ZS, Wang DW, Zhang AM, Ling HQ*State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and DevelopmentalBiology, Chinese Academy of Sciences, Beichen West Road 1, Chaoyang District, Beijing 100101, China*Email: hqling@genetics.ac.cnThe bread-making quality of wheat is strongly influenced by the multiple low molecular weightglutenin subunit (LMW-GS) proteins expressed in the seeds. However, the organization,recombination and expression of LMW-GS genes and their functional mechanism inbread-making are not well understood. Here we report a systematic molecular analysis ofLMW-GS genes located at the orthologous Glu-3 loci (Glu-A3, B3 and D3) of bread wheat usingcomplementary approaches (genome wide characterization of gene members, expression profiling,proteomic analysis). Fourteen unique LMW-GS genes were identified for Xiaoyan 54 (withsuperior bread-making quality). Molecular mapping and recombination analyses revealed that thethree Glu-3 loci of Xiaoyan 54 harbored dissimilar numbers of LMW-GS genes and covereddifferent genetic distances. The number of expressed LMW-GS in the seeds was higher inXiaoyan 54 than in Jing 411 (with relatively poor bread-making quality). This correlated with thefinding of higher numbers of active LMW-GS genes at the A3 and D3 loci in Xiaoyan 54.Association analysis using recombinant inbred lines suggested that positive interactions, conferredby genetic combinations of the Glu-3 locus alleles with more numerous active LMW-GS genes,were generally important for the recombinant progenies to attain high Zeleny sedimentation value(ZSV), an important indicator of bread-making quality. A higher number of active LMW-GSgenes tended to lead to a more elevated ZSV, although this tendency was influenced by geneticbackgrounds. This work provides substantial new insights into the genomic organization andexpression of LMW-GS genes, and the molecular genetic evidence suggesting that these genescontribute quantitatively to bread-making quality in hexaploid wheat. Our analysis also indicatesthat selection for high numbers of active LMW-GS genes can be used for improvement ofbread-making quality in wheat breeding.This work was supported by the Ministry of Science and Technology of China (grants2006AA100102, 2006AA10A105) 88
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 9: Wheat molecular breedingQTL Mapping and Marker Assisted Selection for Some Quality Traits in BreadWheatGupta PK*, Balyan HS, Mir RR, Kumar J, Kumar A, Kumar S, Jaiswal V and Tyagi SMolecular Biology Laboratory, Department of Genetics and Plant Breeding, CCS University, Meerut-250004,India;*E-mail: pkgupta36@gmail.comQTL interval mapping and association mapping were undertaken in our laboratory for a number ofquality traits in bread wheat including pre-harvest sprouting tolerance (PHST), grain proteincontent (GPC) and grain weight (GW). For PHST, we identified several QTL including a majorQTL (QPhs.ccsu-3A.1) that explained >70% phenotypic variation across a number ofenvironments. This major QTL for PHST was introgressed into cv. HD2329 (PHS susceptible),carrying Lr24+ Lr28, using marker-assisted selection (MAS). As a result, seven BC3F3 progenieswere obtained, which had high level of PHS tolerance (PHS score 2-4 on a scale of 1-9) andhypersensitivity towards leaf rust pathotypes. For GPC, a major QTL (Gpc-B1) earlier identifiedby Dr. Jorge Dubcovsky Group at the University of California, Davis, was transferred into six eliteIndian bread wheat genotypes (each with low GPC and carrying Lr24+Lr28). As a result, 14BC3F4 lines carrying Gpc-B1 gene were developed. Ten (10) of these lines had significantly higherGPC, the increment ranging from 0.42% to 2.50% of the total protein content. The selected lineswere advanced to BC3F5 generation, and the progenies homozygous for Gpc-B1 are beingevaluated in replicated field trials over environments. Similarly, for GW, interval mapping led toidentification of 10 QTL, which included 4 major QTL each explaining >20% phenotypicvariation. For GW, association mapping was also conducted, which allowed identification of 9QTL including 7 known QTL that were earlier identified through interval mapping (including twoQTL that were identified by us at Meerut, India and remaining 5 identified elsewhere) and twonew QTL. Marker-assisted selection for improvement of GW using some of the above major QTLis also underway in our laboratory (BC3F1 and BC1F3 generations). The results of the present studydemonstrated successful tagging/mapping of QTL for three quality traits in bread wheat throughboth linkage and association mapping and their subsequent use in marker-aided selection forwheat improvement. 89
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 9: Wheat molecular breedingApplication of MAS for resistance to Fusarium head blight in a wheat breedingprogramW. Cao, G. Fedak, D. Somers, H. Voldeng, M. Savard and A. XueEastern Cereals and Oilseeds Research Centre, AAFC, Central Experimental Farm 960 Carling Ave. Ottawa,ON K1A 0C6. (D.S.) Cereal Research Centre, 195 Dafoe Road, Winnipeg MN R3T 2M9.The objectives of this study were to: 1) determine the effectiveness of marker assisted selection(MAS) relative to conventional visual selection (CVS) for resistance to FHB in a spring wheatbackcross breeding program and 2) develop a white seeded wheat with a high level of FHBresistance using MAS. BW301, a line susceptible to FHB, was crossed with HC374, a lineresistant to FHB and the F1 backcrossed to BW301. A MAS population (MAS BC2F5) wasdeveloped through F2-derived method, while two conventional visual selection populations (CVSBC1F6 and BC2F5) have been developed though single seed descent. Seven lines with all threeFHB QTLs were selected from the MAS population, while the top 10 resistant lines were selectedfrom CVS BC1 and CVS BC2 populations, respectively, based on FHB symptoms in the FHBnursery of 2005. The 27 lines, plus two parents, were further evaluated for FHB resistance in afour replicate field experiment in 2006 and 2007. A combined analysis from two years datashowed that the means for FHB incidence, severity and index and deoxynivalenol (DON)content for the MAS population were 39.2%, 27.3%, 11.2% and 4.6 ppm, for the CVS BC1population the values were 43.0%, 30.8%, 14.4% and 6.1 ppm and; for the CVS BC2population the values were 50.6%, 41.2%, 21.6% and 9.1 ppm. Marker profiles showed thatthe lines in the CVS BC1 population carried from 0 to 2 QTLs whereas none of the lines in CVSBC2 population carried the FHB resistance QTLs. The results confirmed that MAS is moreeffective than CVS for improvement of resistance to FHB in this wheat backcross breedingprogram and indicated that the more backcrosses, the greater the risk of loss of resistance genes inbackcross breeding if the CVS is used. The results also suggested that minor genes have animportant role in determining resistance to FHB in wheat.For development of white seeded wheat with resistance to FHB, Snowbird, a FHB susceptiblewhite hard white seeded wheat, was crossed to Sumai 3 as a female parent. Twenty thousand F2plants were produced and grown in the greenhouse. One thousand and five hundred white seedswere visually selected from the F2 population. This population was advanced to F5 by single seeddescent. At the seedling stage of F5, a MAS was performed for three FHB QTLs on chromosome5A, 3B and 6B. Two hundred and fifty F5 lines were selected with two or three resistance QTLs 90
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京and grown in a FHB nursery in 2008. Fifteen F6 lines were selected based on FHB resistance andagronomic performance. Seed of the 15 lines was increased in the greenhouse in the winter of2009. These 15 lines and two parents Sumai 3 and Snowbird, plus AC Vista as a check wereplanted in the FHB nursery with three replications and in a preliminary yield trial with two reps inthe summer of 2009. The results showed that several white seeded wheat lines had high levels ofresistance to FHB; significantly higher than Snowbird and matured earlier than Sumai 3. Qualityis also improved significantly compared to Sumai 3, based on the Glutomatic test.Keywords: Wheat, Fusarium head blight, marker assisted selection 91
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 9: Wheat molecular breedingGene Function and Modulation of DREB (dehydration-responsive elementbinding protein) Genes from SoybeanChen M, Xu ZS, Li LC, Ma YZ*National Key Facility for Crop Genetic Resources and Genetic Improvement (NFCRI), Key Laboratory of CropGenetics and Breeding, Ministry of Agriculture, Institute of Crop Science, Chinese Academy of AgriculturalSciences (CAAS), Beijing 100081, China;* E-mail: mayouzhi@yahoo.com.cnThe transcription factors DREB specifically bind the dehydration- responsive element/C repeat(DRE/CRT) cis-acting element and control the expression of many stress-inducible downstreamgenes in plants. By screening of drought-induced soybean cDNA library, we isolated four cDNAfor DREB homologs: GmDREB1, GmDREB2, GmDREB3 and GmDREB5. The expression ofGmDREB2 was induced by drought, high-salt (200 mmol/L NaCl), low temperature (4℃) stressesand ABA (200 umol/L) treatment, and GmDREB3 was induced by low temperature, and theGmDREB5 was induced by high-salt and drought stresses. Gel mobility shift assay showed thatthese four genes specifically bound to DRE/CRT element in vitro. The transcript activating assayshowed that those genes specifically activated expression of downstream genes fused with thepromoter containing three randomly repeated copies of the wild-type DRE/CRT sequence.Functional analysis indicated that expression of those GmDREBs enhanced tolerance to differentstresses such as drought, high salt or low temperature, suggesting that those GmDREBs arepotentially useful for improving stresses tolerance of crops. In addition, to elucidate themodulation of DREB transcription factors in the process of translation, some genes encodinginteractive proteins with these DREB proteins were isolated from soybean using yeast two hybridsystem. To date, some interactive proteins such as E2 lingase, TPR repeat containing protein wereisolated from soybean. Their interactive activity were proven using Put-down method.Furthermore, analysis of the GmDREB3 promoter elucidated its cold-induced modulation: apromoter fragment was involved in response to cold stress and its effect was detected for 1 h aftertreatment, and transcriptional repressor appeared to impair this response by binding to acis-element in anther region 24 h after cold stress. These results are useful for elucidatingregulative mechanism of these DREB proteins under various stress condition. 92
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 9: Wheat molecular breedingGenomic Distribution of Quantitative Trait Loci (QTL) for Yield andYield-related Traits in Common Wheat (Triticum aestivum)Zhang LY1, Liu DC1, Guo XL2, Yang WL1, Sun JZ1, Wang DW1 and Zhang AM1*1 The State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics andDevelopmental Biology, Chinese Academy of Sciences, Beijing 100101, China2 Biological College, China Agricultural University, Beijing 100193, China* E-mail: amzhang@genetics.ac.cnA major objective of quantitative trait locus (QTL) studies is to find genes/markers that can beused in breeding programs via marker assisted selection (MAS). We surveyed the QTLs for yieldand yield-related traits and their genomic distributions in common wheat in the available literature.We then performed a meta-QTL (MQTL) analysis to identify the major and consistent QTLs forthese traits. In total, 55 MQTLs were identified, of which 12 significant MQTLs were located onwheat chromosomes 1A, 1B, 2A, 2D, 3B, 4A, 4B, 4D and 5A. Our study showed that the geneticcontrol of yield and its components in common wheat involved the important genes such as Rhtand Vrn. Furthermore, several significant MQTLs were found in the chromosomal regionscorresponding to several rice genomic locations containing important QTLs for yield related traits.Our results demonstrate that meta-QTL analysis is a powerful tool for confirming the major andstable QTLs and refining their chromosomal positions in common wheat, which may be useful forimproving the MAS efficiency of yield related traits. 93
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 10: Molecular breeding platform and new technologiesThe Integrated Breeding Platform: Vision and PracticeMcLaren GGeneration Challenge Program c/o CIMMYT Apdo Postal 6-641, 06600 Mexico D.F. Mexico.Email:g.mclaren@cgiar.orgA corporate-led ‘gene revolution’ has boosted crop productivity and resilience in developedcountries. The Integrated Breeding Platform (IBP) is a publicly funded, collective effort to do thesame in developing countries, especially for smallholders in drought-prone environments.The Generation Challenge Programme (GCP) of the Consultative Group on InternationalAgricultural Research (CGIAR) is building the IBP with the collaboration of 14 initial ‘user cases’.These breeding projects – for eight crops in 16 African and Asian countries – help ensure that IBPdevelopment remains driven by demand. As the IBP gradually opens up to other users starting in2012, the initial user cases will advertise its utility as a one-stop shop for modern marker-assistedbreeding technology. Full service for all eligible requests is scheduled for 2014.The IBP has three components: a web-based portal and helpdesk, an open-source informationsystem, and breeding and support services. The service component is the most advanced, with IBPbrokerage of molecular marker laboratory services well underway and its informatics system formanaging breeding data available.The IBP is doing a lot of training and cleaning historical data to make it more useful. Otherpriority services under development with user cases are design and analysis for experiments,providing to users phenotyping sites and screening protocols, and assistance with drawing upbreeding plans.One project objective towards building the information system is to make available to usersexisting tools for managing pedigree information, field data and laboratory information. Inaddition, the IBP is developing a suite of decision-support tools that will be linked into aconfigurable workflow system, as well as a crop information network..Scheduled for inauguration by the end of 2010, the portal will be the online window throughwhich users select and download tools and instructions, order materials, and procure laboratoryservices. The portal’s helpdesk will facilitate its use and ensure access for users who cannotefficiently use the web interface by providing the elements they need via email, compact disc andother media.The project is funded by the Bill & Melinda Gates Foundation, United Kingdom’s Department forInternational Development, and European Commission until late 2014. Its business plan envisionsthe platform carrying on as a CGIAR service supported in part by annual fees paid by members ofa formal IBP user community. 94
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 10: Molecular breeding platform and new technologiesOptimization of NGS- based SNP discovery approaches for facilitatingmolecular breeding in orphan crop speciesThakur V1, Ruperao P1, Amindala B1, Farmer AD2, Jayashree B1, Studholme DJ3, May GD4, Jones JDG3, ShahT1, Varshney RK1,4,*1 International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Patancheru 502324, AndhraPradesh, India;2 National Center for Genome Resources (NCGR), New Mexico, Santa Fe, 87505 USA;3 The Sainsbury Laboratory (TSL), JIC, Norwich Research Park, Norwich NR4 7UH, UK;4 CGIAR- Generation Challenge Program (GCP), c/o CIMMYT, 06600 Mexico DF, Mexico* Email: r.k.varshney@cgiar.orgNext-generation sequencing (NGS) approaches are generally used for re-sequencing individualsfor discovery of variants (SNPs) for those species where a reference genome sequence is available.Several tools are available for analyzing the NGS data that can align the short sequence tags(SSTs) and identify SNPs in the individuals in relation to genotype for which reference genomesequence is available. Despite much advancement in genomics, several crops like chickpea,pigeonpea still lack appropriate genomic resources such as molecular markers, genetic maps thatare critical for undertaking molecular breeding. By using chickpea (Cicer arietinum) as anexample, it has been demonstrated that NGS technologies together with appropriate sequenceanalytical approach, can facilitate development of SNP markers for enabling molecular breedingin called ‘orphan crop’. In this context, a chickpea transcriptome assembly was developed basedon Sanger EST (21,491) and 454/FLX transcript reads (435,018). In parallel, 15.66 milliontranscripts reads (SSTs) for ICC 4958 and 22.09 million SSTs for ICC 1882 were generated byusing Illumina sequencing approach. For identification of robust set of SNPs between these twogenotypes, a consensus calling approach, based on depth and frequency of variant nucleotide wasinvestigated by using four commonly used tools Alpheus, MAQ, NovoAlign and SOAP. Anexamination of consistency in SNP prediction showed relatively large variation. For instance, inabove dataset, a total of 5,358 SNPs were identified by using above mentioned four tools;however, 78.6% SNPs were unique to the four tools and only 2.2% SNPs were common with allfour tools. On the other hand upto 4.1% SNPs were found common with two tools and amaximum of 3.5% SNPs with three tools. Based on validation of in silico predicted SNPs withdifferent tools, our study suggests that identification of reliable or robust set of SNPs is stillchallenging in crops like chickpea, without a reference genome sequence, read depth criteria isprobably the best parameter that can be used to call SNPs with higher precision. These resultsincluding optimized parameters will be discussed in details in the presentation. 95
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 10: Molecular breeding platform and new technologiesISMAB: A Data Visualization and Decision Support Tool for CropImprovementShah T*, Balaji J, Tuteja R, Nair S, Nepolean T, Hash CT, Punna R, Davenport G, McLaren G and Hoisington D*Email: tm.shah@cgiar.orgData visualization is critical in agricultural research, because of the large amount of complex datagenerated in genomics and crop breeding. A major challenge in crop improvement programs is ininterpreting and decision making from large data sets that combine information on genotypes,phenotypes and environments. Visual analytics allows scientists to view their information in agraphical environment, where they can more easily understand the relationships underlying thedata, identify trends and patterns, and make discoveries based on information visualization. Weare focusing on visualization of Marker-Assisted Breeding (MAB) experiment datasets, to providea workbench that can integrate information from various experiments, across generations andidentify the best individuals by visual analysis.We describe the development of an Information System for Marker-Assisted Breeding (ISMAB),which can be used as a tracking and visualization system for MAB. The first component, theMolecular Breeding Design Tool (MBDT), has been developed that assists in the selection ofsuitable parents, provides visual integration of genotypic, map and Quantitative Trait Loci (QTL)data and allows rich client-based interactivity and manipulation. 96
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 10: Molecular breeding platform and new technologiesBring genomic data to breeders: what we expect from IBP to help futurebreedingLiang CZInternational Rice Research Institute, DAPO Box 7777, Metro Manila, PhilippinesEmail:cliang@cgiar.orgData integration is a hot area in current bioinformatics research. It is the foundation of system biology toachieve a holistic, quantitative, and predictive understanding of complex biological problems through iterativeanalysis of prediction of mathematical models and experimental results. Nevertheless, numerous efforts havebeen done to integrate ‘-omics’ data with limited success. The major obstacles to achieve successful dataintegration include: 1) highly hierarchical data structure, usually forming network interleaved with each other, 2)dynamic data source and type, 3) lack of standardization in data format and nomenclature, 4) large data columnbut incomplete, 5) data error. In future plant breeding we need the most efficient usage of genetic variation,which call for prediction of phenotype from genotype (ie, genomics-assisted breeding). The complex breedingdata demands continuous development of new data management and visualization tools. The utilization of thegenomic data through high density molecular markers in breeding line prediction/selection requireswell-developed statistical models/computation algorithms (data mining tools) to be integrated with the datamanagement/visualization system.The GCP/BMGF funded Integrated Breeding Platform (IBP) aims to build a one-stop-shop for informationretrieval and analytical support on various aspects of breeding activities. A fully fledged IBP framework willhelp integrate phenotypic, genotyping, genomic and comparative genomic data, to improve the precision andefficiency of predicting phenotypes from genotypes, and thus increase breeding efficiency by deceasingbreeding cycles or increasing selection precision. At IRRI we aim to provide a fully integrated rice germplasminformation system on the top of IBP development to support the modern rice breeding strategies. Some currentand future activities include: 1. develop data curation and management tools, processing pipelines for association of heterogeneous data 2. integrate internal or external genotyping, phenotypic, genomic and comparative genomic data (eg, IRIS and Gramene) 3. develop and integrate web-based and desktop visualization tools, eg, pedigree viewer – for parental selection, and chromosome segregation viewer – to visualize the segregation of chromosome fragments in progeny lines derived from their parent, thereby identifying crossover events in breeding population 4. develop and integrate web-based and desktop data query and reporting tools for data query, download and graphical report 5. integrate breeding decision-making supporting tools, including prediction and simulation tools 97
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 10: Molecular breeding platform and new technologiesDevelopment and Optimization of the 50K Infinium Chip for Maize DiversityAnalysisGanal M1*, Polley A1, Durstewitz G1, Thompson M2 and Hansen M21 TraitGenetics, Gatersleben, Germany; 2Illumina, San Diego, USA*Email: ganal@traitgenetics.deIn a collaborative effort between multiple partners (Illumina, USDA, Syngenta, INRA andTraitGenetics), an Infinium array has been designed that consists of more than 50000 maize SNPs.Through the analysis of a first set of maize lines an optimized cluster file has been generated thattakes into account the difficulties caused by the high diversity of maize lines and the fact thatmaize is an ancient tetraploid species. Data will also be presented regarding the type, quality andlevel of polymorphism of the SNPs on the array in a set of mainly European maize breeding linesand we will also present some data regarding the potential use of the array to generate highdensity genetic maps with many thousands of markers. This array offers a tool to analyse diversityin the entire maize genome or chromosomal segments of the maize genome. Moreover, it offersthe potential to perform diversity studies in many of the maize genes that contain SNP markers onthe array. 98
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 11: Germplasm and genetic diversityCore collection-based genomic stocks in wheatJia JZ*, Kong XY, Gao LF, Zhang XY, Zhou RH and Dong YCInstitute of Crop Sciences, CAAS, Beijing 100081, China* Email: jzjia@mail.caas.net.cnOne core collection (CC) and one mini-core collection (MCC) consisting of 1,160 and 231accessions, respectively, were assembled from a collection of 23,090 Chinese wheat accessions.The CC and MCC collections captured more than 90 and 70% of the genetic diversity present inthe initial collection, respectively. Four hundreds and eighty-seven accessions including the MCC,accessions introduced from across the world and synthetic wheats were crossed with Chinesecommercial leading varieties followed by selfing and backcrossing. Consequently, 386populations of recombination inbred lines (RILs) and 411 populations of introgression lines (ILs)were developed. The MCC and its derived RILs and ILs are identified as MCC-based genomicstocks. The MCC-based genomic stocks have been deployed for gene discovery through linkageand association analysis, allowing us to detect hundreds of new agronomically importantgenes/QTLs, that affect yield components, grain quality, stress tolerance and disease resistance.Hundreds of elite lines selected from the ILs and RILs have been used as parents in Chinese wheatbreeding programs and some of them have been released as new cultivars. Our results havedemonstrated that the MCC-based genomic stocks are valuable for both functional genomicsstudies and breeding, and are expected to contribute significantly to wheat improvement in the nearfuture. 99
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 11: Germplasm and genetic diversityHigh-throughput SNP genotyping of a subset of lettuce landraces for geneticdiversity assessmentHu JG*, Kwon SJ and Hellier BUSDA-ARS Western Regional Plant Introduction Station, Washington State University, Pullman, Washington,USA.* Email: jinguo.hu@ars.usda.govLettuce is one of the most popular vegetables grown commercially worldwide. It ranks as one ofthe top ten most valuable crops in the U.S. with an annual value of over two billion dollars since2004. The USDA lettuce germplasm collection is maintained in seed form in the WesternRegional Plant Introduction Station at Pullman, WA. We are in the process of fingerprinting thewhole collection of approximately 1,600 accessions with the high-throughput SNP genotyping forgenetic diversity assessment. We used the service provided by the DNA Technologies CoreFacility of UC Davis Genome Center for SNP genotyping. The oligo pools (OPAs) targeting 384SNP loci were designed from the high quality EST (expressed sequence tag) sequences generatedby the US Compositae Genome Project (CGP) headed by Dr. Richard Michelmore of UC Davis.SNP genotyping was carried out by using the Illuminas custom VeraCode GoldenGateGenotyping Kits which can simultaneously genotype 384 loci in a single well of a standard96-well microplate. The scanned data from BeadXpress Reader were analyzed with theGenomeStudio software to generate genotype data for individual accessions. Our initial resultfrom 455 accessions demonstrated that this technique is very powerful in revealing geneticvariation. Three hundred and thirty-five or 87% of the 384 SNP produced polymorphic genotypedata and each of the 455 accessions could be discriminated by the dataset. Detail results from asubset of lettuce landrace germplasm will be presented. 100
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 11: Germplasm and genetic diversityGenetic Diversity Studies on Cool Season LegumesXuxiao Zong1*, Shumin Wang1, Jin Liu1, Rebecca Ford2, Bob Redden31 Institute of Crop Sciences/ National Key Facility for Crop Gene Resources and Genetic Improvement, ChineseAcademy of Agricultural Sciences, Beijing 100081; 2 BioMarka, Faculty of Land and Food Resources,Melbourne University, Victoria, Australia 3010; 3 Australian Temperate Field Crops Collection, GrainsInnovation Park, The Department of Primary Industries, Private Bag 260, Horsham, Victoria, Australia 3401.*Email: zongxx@mail.caas.net.cnTotally 2120 accessions including cultivated and wild species from 8 botanical unites under Pisumgenus which originated from 67 countries and China (28 provinces), were examined using 21polymorphic SSR primer pairs in ICS of CAAS. Significant difference has been identifiedbetween genetic resources of within and outside China in genetic diversity. Three gene pools havebeen detected among genetic resources under Pisum Genus. Genetic resources from Springsowing area of China differed significantly to that from winter sowing area of China in general.Genetic resources from Inner Mongolia and from Sha’anxi provinces of China are unique. Fourgene pool clusters were detected under Pisum genus by using PCA analysis. The UPGMAclustering results generally support the PCA clustering results. Gene pool “fulvum”mainlyconsisted of wild species Pisum fulvum, gene pool “abyssinicum” mainly consisted of P. sativumssp. abyssinicum, and genepool “arvense” mainly consisted of P. sativum ssp. sativum var.arvense. While gene pool “sativum” were composed by five botanical taxon units, they are P.sativum ssp. asiaticum, P. sativum ssp. elatius var. elatius, P. sativum ssp. transcaucasicum,P.sativum ssp. elatius var. pumilio and P. sativum ssp. sativum var. sativum. There weresignificant differences among most botanical groups under Pisum genus, with clear separation offour gene pools for genetic diversity structure. The research results partially support the traditionalbotanical taxonomy under Pisum genus.AFLP primer pairs were applied to examine 240 winter genotypes and 216 spring genotypes offaba been for genetic diversity studies in ICS of CAAS. 2D-, 3D-PCA graphs shown significantdifference between genetic resources of the Chinese local and from outside China in geneticdiversity. UPGMA shown greater difference in genetic diversity between spring sowing geneticresources and winter sowing genetic resources of Chinese local, to that between winter sowingaccessions from within China and from outside China. Both UPGMA and 3D-PCA shown Yunnanaccessions are unique in Chinese winter faba accessions. 3D-PCA graph shown significantdifference among continental originated groups of spring sowing faba bean genetic resources.ICARDA faba bean is unique. Three clusters were identified by NJ dendrogram. Four populationswere detected by Structure analysis: the population Pop 1 approximated to Sub-cluster A1, Pop 2to Sub-cluster A2 and Sub-cluster A3, Pop 3 to Sub-cluster A4 and Pop 4 to the combination ofCluster B and Cluster C. UPGMA dendrograms shown spring faba beans from Europe and Africa 101
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京are similar; Spring faba beans from Asia and ICARDA are similar; Spring faba beans fromAmerica are unique.440 lentil accessions were examined based on 14 SSR primer pairs in ICS of CAAS. Althoughrich genetic diversity existed in Chinese local lentil genetic resources, the diversity level of globallentil genetic resources is higher. Eight populations were identified by Structure analysis. Chineselocal populations are similar in genetic diversity; populations from outside China are diversifiedin genetic diversity. The results doubt the hypothesis “The species Lens culinaris was introducedto China from Indian from beginning”, and raised the possibility that Chinese lentil comes fromEurope. 102
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 11: Germplasm and genetic diversityThe genetic diversity, structure and classification of rice germplasm in ChinaZhang DL, Zhang HL, Li JJ, Li ZC*Dept of Plant Genetics and Breeding, China Agricultural University, Beijing 100193, China*Email: lizichao@cau.edu.cnChina is one of the largest centers of genetic diversity of common wild rice (CWR) (Oryzaruffpongon Griff.) and Asian cultivated rice (ACR) (oryza sativa L.), and one of the centers oforigin of ACR. There are about 70,000 accessions of rice germplasm in Chinese Rice Genebank.In terms of phenotype and SSR genotype, during the domestication process (from CWR to ACR)one third of the variations in CWR lost, and just 7.6% of the variations in ACR generated newly,and nearly 20% of the variations in local rice varieties (LRV) have not been used in commercialrice varieties. These results indicate that wild rice and local rice varieties (especially the former)are very important sources of genetic variation in improving the rice varieties.The genetic structure of CWR suggests that CWR has diverged into two ecotypic populations (anindica-like type and a japonica-like type), and further seven geographical populations. The naturalselection due to different climates, the geographical isolation and the water systems contributed tothe formation of the genetic structure. The genetic structure of ACR confirmed the primarydifferentiation between indica and japonica, but an intra-subspecific differentiation patterndifferent from the previous opinion. Japonica appeared more distinct differentiation among threesoil-watery ecotypes, whereas indica was more clearly subdivided by three seasonal ecotypes.Within each soil-watery ecotype and seasonal ecotype, various geographical-ecotypic populationswere detected. Different environments were the primary factor of the differentiation of twosubspecies, whereas the different cropping systems and geographic isolation imposed on twosubspecies led to their further differentiations. Based on the above results, we suggested a newclassification system of cultivated rice, and we screened a set of SSR markers to distinguishdifferent populations in the system.It has been approved that establishment of core collection (CC) offers one effective approach tothe conservation and use of the genetic diversity in a large germplasm resources. In establishmentof core collection of Chinese rice germplasm resources, we recommend two principles on thedetermination of the CC size and a hierarchical CC system. In the system there are different coresets with different population scales and genetic diversities, such as C2000, C1500, CC, C500 andmini core collection (MCC), and thus allow a flexible use of genetic resources. The CC comprises1.7% (932) of the accessions in the basic collection, and retains more than 85% of both the SSRand phenotypic variations; the MCC comprises 0.3% (189) of the accessions in the basiccollection, retains 70.65% of the SSR variation and 76.97% of the phenotypic variation. Beingabundant in diversity and practical in scale, MCC is an ideal population of gene discovery andbreeding. On the one hand, we have developed approximately 60,000 introgression lines and 103
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京isogenic lines based on MCC. On the other hand, using 300 genome-wide SSR markers we havefinished the association mapping of more than 50 traits phenotyped at three locations (Beijing,Hangzhou, and Hainan) for 2 years.In association mapping, the GLM model integrating with population structure (Q) performs betterthan the EMMA model integrating with both population structure and relative kinship (Q+K) forall traits. Approximately one third of the markers associated with more than one trial. It impliesthat the expression of most genes is induced by different environment factors. Some markersassociate with more than one traits and thus may contribute to the correlation among traits andpleiotropism. Take yield as an example. We detected a complex genetic network controlling riceyield and each yield component. Finally, several genomic regions were recognized as the target toidentify genes related to rice yield; and a set of materials with novel high-yield genotypes werepicked out as the potential parents to develop super-rice.In conclusion, these efforts will help us to determine the extent of natural variations in rice and themolecular mechanisms underlying complex traits. 104
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 11: Germplasm and genetic diversityPlantain (Musa sapientum L) Molecular diversity Reveals Narrow Genetic Baseof Local Ghanaian AccessionsMarian D. Quain*, and Beloved DzomekuCSIR-Crops Research Institute, Ghana* Email: marianquain@hotmail.comPlantains are important sources of high-calorie energy in Ghana. They are also of greatsocio-economic importance in the country. Plantains are also very important sources of ruralincome. However, plantain production has been saddled by several constraints among whichinclude pests and diseases and lack of improved technologies. As regards to genetic study,morphological characterization has identified about 100 local accessions to exist in Ghana.However, no molecular characterization has been done on plantains. Characterization based onmorphologic characteristics alone may be limited since the expression of quantitative traits issubjective to strong environmental influence. Alternatively, molecular characterization techniquesare capable of identifying polymorphism represented by differences in DNA sequences. Theobjective of this research was therefore to conduct molecular characterization of Ghanaian localaccessions of plantain. This study project sampled 38 accessions of plantains in Ghana from theAshanti and Eastern region. Sampling covered all known morphologically categories. Molecularcharacterization was carried out on the assembled germplasm. Simple Sequence Repeats (SSRs)were used to assess diversity. The SSR marker revealed that 24% of the plantain accessions have agenetic distance of zero units. “Asamienu” was most distantly related to all the other varieties. Anintroduced hybrid Fhia 21 was also distantly related to all the local accessions, however, it wasfound to be closely related to Apantu Red and Apantu Dichotomy. “Orishele Eumusa” an AABPlantain of Nigerian origin was related at distance Zero to some of our local varieties. Whereas“Yade Eumusa” an AAB Plantain of Cameroon origin was closely related to some of the localaccessions. The dendogram generated two major clusters. “Asamienu” was distantly related to allassessions whereas FHIA 21 closely related to two of the local Apantu collections. The SSR datapointed out that the plantain accessions in Ghana are very closely related and hence breedingprograms need to focus on broadening genetic base. This is the first report on the use of molecularmarker tools to carry out diversity studies on plantains in Ghana. The information generated willbe used as a document to encourage breeders to work towards broadening genetic diversity inplantains. 105
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 11: Germplasm and genetic diversityThe strategy and potential utilization of temperate germplasm for tropicalgermplasm improvement-a case study in maize (Zea mays. L)Wen WW1, Guo TT2, Tovar VHC1, Yan JB1, 2, Taba S11 International Maize and Wheat Improvement Center (CIMMYT), Apartado Postal 6-640, 06600 Mexico, DF,Mexico;2 National Maize Improvement Center of China,China Agricultural University, Beijing 100193, China.Organization of maize germplasm into genetically divergent heterotic groups is needed forbreeding high yielding hybrids by exploitation of heterosis. In this study, 94 CIMMYT inbredlines (CML) and 54 US Germplasm Enhancement of Maize (GEM) lines were assembled andcharacterized using 1,266 SNPs with high quality. Based on principle component analysis (PCA),GEM and CML lines were clearly separated. There were two groups of GEM lines classified byPCA that seemed to correspond to the stiff stalk (SS) and non-stiff stalk (NSS) heterotic groupsbred by GEM. CML lines did not form obvious subgroups by PCA. Each of 1,266 SNP allelefrequency differed in GEM and CML. 3.6% alleles (46/1,266) of CML were absent in GEM, and4.4% alleles (56/1,266) of GEM were absent in CML. CML heterotic group A with GEM heteroticgroup SS and CML heterotic group B with GEM heterotic group NSS were crossed and developed654 F1. Association mapping was performed based on both 148 CML, GEM lines and 654 F1s forseven agronomic traits at two locations. For plant height (PH) and anthesis time (AT), genomicestimated breeding values (GEBVs) for a testing set of 215 F1s were predicted based on thetraining data of 430 F1s using a best linear unbiased prediction (BLUP) method. The accuracybenefitted from the adoption of markers associated with QTLs for both traits. However, it does notnecessarily increase with the raise of associated marker density. High correlations between actualand predicted phenotypic values of F1 indicated the possibility to choose high performing parentalcombinations of phenotypic traits using SNP markers. It is hoped that we can enhance the tropicalmaize germplasm by incorporating temperate germplasm more efficiently, through genome wideselection. Clear heterotic patterns of the GEM lines and unique alleles can facilitate tropical maizehybrid breeding. 106
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 12: Molecular breeding in legumes and trees cropsConcentration of genetic diversity for gene discovery and broadening geneticbase of modern cultivar in soybeanQiu LJInstitute of Crop Science, Chinese Academy of Agricultural Science, Beijing 100081. ChinaEmail: qiu_lijuan@263.netSoybean is the Chinese traditional food source of plant protein and oil. It has been cultivatedover 4500 years. More than 1500 cultivars have been developed and released in China since1920. These cultivars can be traced back to ~300 ancestral lines, which take about 1% of the over30000 accessions reserved in National Crop Genebank in China. In order to broadening thecultivar genetic base with selected divergent genetic resources, we evaluated Chinese soybeangermplasm with passport data, morphological traits and chosen a set collection as primary corecollection (Qiu et al., 2003), and submit for diversity assessment with SSR markers (Li et al.,2008). Then a subset of accessions was selected as core collection (Wang et al, 2006) andfurther concentrated to a mini core collection. The representative of the mini core collection wasused for survey gene diversity of newly cloned determinate growth gene GmTFL1 (Tian et al.2010) as well the other useful traits and genes. Meanwhile, these accessions had beencooperated into soybean breeding program as donors to cross and backcross with the localcultivars (Qiu et al., 2009) for broadening genetic base of modern cultivars. 107
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 12: Molecular breeding in legumes and trees cropsDevelopment and application of genomic resources for molecular breeding ingroundnut (Arachis hypogaea L.)Varshney RK1,2*, Gowda MVC3, Radhakrishnan T4, Pandey MK1, Gautami B1, Sujay V1,3, Koppolu R1,Senthilvel S1, Vadez V1, Nigam SN1, Upadhyaya HD1, Isobe S5, He G6, Bertioli DJ7, Knapp SJ8, Cook DR91 International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Patancheru, Greater Hyderabad502324, India; 2 Generation Challenge Programme (GCP), c/o CIMMYT, 06600 Mexico DF, Mexico;3 University of Agricultural Sciences (UAS)-Dharwad, 580005, INDIA; 4 Directorate of Groundnut Research(DGR), Junagadh, 362001, Gujarat, India; 5 Kazusa DNA Research Institute (KDRI), Kisarazu, Chiba 292-0818,Japan; 6 Tuskegee University, Tuskegee, AL 36088, USA; 7 Universidade Católica de Brasília (UCB), Brasília,Brazil; 8 The University of Georgia, Athens, GA 30602, USA; 9University of California-Davis, CA 95616, USA.*Email: r.k.varshney@cgiar.orgGroundnut (Arachis hypogaea L.) is an important oilseed crop grown extensively throughout thesemi-arid tropics of Asia, Africa and Latin America. Genetic yield potential has been adverselyaffected due to terminal drought and foliar diseases. Marker-assisted selection offers an importanttool to enhance tolerance/resistance to above stresses which are difficult through conventionalbreeding. However, a very limited amount of genomic resources were available until recently.As a collaborative effort with several partners, large scale genomic resources such as simplesequence repeat (SSR) and Diversity Array Technology (DArT) markers have been developedrecently. These markers together with others available in public domain or accessible throughcollaborators are being/have been used to detect polymorphism between parental genotypes offive mapping populations namely TAG 24 × ICGV 86031, TAG 24 × GPBD4, TG 26 × GPBD 4 ,ICGS 44 × ICGS 76 and ICGS 76 × CSMG 84-1. In general, 6-10% polymorphism has beenobserved with the SSR markers tested. Genotyping of polymorphic markers has facilitateddevelopment of genetic maps with moderate marker density (83-191 per cross) for respectivemapping populations. Moreover, a consensus genetic map comprising 290 SSR loci spanning 20linkage groups has been developed based on 3 mapping populations of cultivated groundnut.Detailed QTL analyses for drought component traits and disease resistance has detected a total of125 QTLs. For majority of traits studied, QTLs identified had small phenotypic effect. However,one major QTL, contributing up to 54.4% phenotypic variation for rust resistance has beenidentified and validated among diverse germplasm/alternate mapping populations. This enabledinitiation of marker-assisted introgression of this major QTL into elite groundnut cultivars. Insummary, molecular markers, genetic maps and QTLs will be of great help for facilitatingmolecular breeding and improving crop productivity of cultivated groundnut. 108
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 12: Molecular breeding in legumes and trees cropsSearching for QTLs to enhance yield stability in durum wheatMaccaferri M1, Sanguineti MC1, Graziani M1, Bassi F, Corneti S1, Stefanelli S1, Demontis A2, Massi A2, AmmarK, Paux E3, Feuillet C3, Tuberosa R1*1 Dept. of Agroenvironmental Science & Technology (DiSTA), U. of Bologna, 40127 Bologna, Italy; 2SocietàProduttori Sementi di Bologna PSB, 40050 Argelato, Bologna, Italy3 CIMMYT, 56134 Texcoco, Mexico;4 UMR INRA-UBP 1095, Domaine de Crouelle 234, 63100 Clermont-Ferrand, France.*E-mail: roberto.tuberosa@unibo.itDurum wheat (Triticum durum Desf.) is an important staple in the diet (e.g. cous-cous, pasta,bread, etc.) of Mediterranean countries, where drought is the main stress curtailing yield. AtDiSTA, we have undertaken a long-term program to identify and eventually clone genes/QTLsable to mitigate the negative effects on grain yield of drought. To this end, a two-prongedapproach based on both linkage and association mapping has been instrumental to identify anumber of relevant major loci. In particular, we have identified two major QTLs on chrs. 2BL and3BS that influenced leaf senescence, kernel weight and grain yield in a RIL population (Kofa ×Svevo) tested across 16 environments characterized by a wide range of soil moisture regimes(Maccaferri et al. 2008; Genetics 178:489-511.). For the fine mapping of these two QTLs nearisogenic lines (NILs) have been derived. Field trials conducted with the NILs of the chr. 3BS QTL(QYld.idw-3B) have confirmed the effects originally described for this QTL. The analysis of thesequencing data obtained by Feuillet and coworkers (TriticeaeGenome project) have assignedQYld.idw-3B to a BAC contig (3156 kb-long) that contains 41 ORFs.In parallel to biparental mapping, association mapping has highlighted the role of additional QTLsfor yield in a panel of elite durum accessions grown across different water regimes (Maccaferri etal. 2010; submitted). To streamline the genetic profiling of mapping populations and panelssuitable for association mapping, in collaboration with KeyGene we have undertaken thediscovery of SNPs to implement a Golden Gate Illumina based assay. So far, we have identifiedand validated 768 SNPs, 159 of which have been mapped in two RIL populations. Eventually, thecloning of major QTLs able to influence the sustainability of durum wheat production, willprovide the means for a more targeted and accurate mining (e.g. EcoTILLING) and manipulationof valuable alleles towards the release of improved cultivars. 109
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Concurrent session 12: Molecular breeding in legumes and trees cropsGenomics tools to aid cassava breeding for drought toleranceRabinowicz P1, Myburg Z2, Luo MC3, Lopez-Lavalle AB4, Okogbenin E5, Fregene M6, Morris J2, Rey C71 Institute for Genome Sciences, Univerity of Maryland School of Medicine, USA;2 University of Pretoria, South Africa;3 Univeristy of California, Davis, USA;4 International Center for Tropical Agriculture, Colombia;5 National Root Crops Research Institute, Nigeria; 6Danforth Plant Science Center, MO, USA; 7University of theWitwatersrand, South Africa.Cassava is a major staple crop in developing countries, especially in sub-Saharan Africa whereenvironmental conditions are often extreme due to severe drought. In order to generate genomictools to aid cassava breeding for drought tolerance we are developing a genome-wide singlenucleotide polymorphism (SNP) resource for cassava. SNP markers were developed from twosources: i) BAC-end sequences from a cassava BAC fingerprint map, and ii) gene modelsannotated in the cassava genome released by Roche and the US-DOEs JGI. The cassavafingerprint physical map that we generated contains nearly 60,000 BAC clones assembled into2,104 contigs and 5,054 singletons. A minimum tiling path (MTP) derived from this map spans710 Mbp and includes nearly 7,000 clones. 384 SNPs were derived from BAC-end sequencesspread across the MTP and another 384 SNPs were derived from gene models from JGIs cassavagenome annotation. Genotyping of these SNPs will be carried out in a mapping populationderived from a cross between cassava lines with contrasting drought tolerance phenotypes. Thesame population will be phenotyped in Brazil and Colombia to conduct QTL analysis for droughttolerance in cassava. We will also leverage the castor bean genome sequence for cassavagenomics research. 110
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京 Posters 111
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京S 1.01 - Analysis of DNA methylation patterns of different physiological age from PhyllostachysPubescens based on MSAPGuo GP, Yuan JL, Yang K, Gu XP *(guxp@fy.hz.zj.cn)Research Institute of Subtropical Forestry, CAF, Fuyang 311400, Zhejiang, ChinaThe research of DNA methylation of woody plant indicated that there is may be a close relationship with the ageof plants. And we will have a further study for the relationship through research the tender leaves ofPhyllostachys pubescens of three different ages (5 years, 30 years and more than 60 years),using a newtechnique called methylation sensitive amplified polymorphism (MSAP). After a conventional statistics, wecome to a conclusion: DNA methylation level was ascended with the proceeding of senescence. The rate ofDNA methylation in amplified loci was 24.47%, 28.23% and 32.13%, and the total rate of that was 16.57%,19.41% and 21.23% respectively and after a variance analysis about full methylation level, we find themethylation- sensitive amplied polymorphism show an stable upward tendency with the raise of age ofPhyllostachys pubescens and it is very salient in three chosen age class. The ANOVA confirmed there is sixpairs (E3/HM2 (X1), E3/HM6 (X2), E3/HM7 (X3), E4/HM5 (X4), E4/HM6 (X5) and E5/HM5 (X6) of primersamong the thirtyfive pairs of primers we selected have a marked influence of physiological age on DNAmethylation, and the differences between different physiology age up to extremely significant.. The futheranalysis using Main points of law indicate that there is a aggregative indicator which can show the grossinformation content of DNA methylation level of the Phyllostachys pubescens, and the expression is Y1 = 0.173X1+0.172 X2+0.172 X3+0.178 X4+0.176 X5+0.180 X6. Then, we test more ages Phyllostachyspubescens,consist of 2 years, 6years, 13 years, 18 years, 31 years, and more than 61 years. Take advantage ofSPSS conduct a curve fitting in the ration of methylation of different physiology age and show an incrementalrendering. Besides, the relatons between physical age and DAN methylation level was refined. That is:y=0.542x2-0.003x+17.999, R2=0.970.S 1.02 - Mapping of Nuclear Male-sterile Genes ms14 using SSR markers in CottonHu BM , Hu L and Wang PZXinjiang Condy Seed Science & Technology Development Co. Ltd, Urumqi, 830011.* Email: Condywpz@126.comNuclear male sterility (NMS) is a very important character for cotton breeding and genetics programs.Identification of linked molecular markers with NMS will greatly facilitate breeding for this trait. Among all ofthe Nuclear male-sterile genes, only Dong-A (ms14) and ms5ms6 lines were utilized successfully in hybridproduction of cotton. Dong-A (ms14), a recessive NMS line developed from spontaneous mutation in uplandcotton, has been applied most widely and played an increasing role in hybrid cultivar development in China. Anintraspecific F2 population comprised of 180 individual plants was developed by crossing a cultivar of Kang A(Dong-A derived Lines, Gossypium hirsutum L.) to a cultivar of 601588 (Gossypium barbadense L.). Base onthis population the ms14 gene was mapped on chromosome 2 and closely linkaged with BNL3971 (simplesequence repeat markers) within a genetic distance of 16.7 cM. The BNL3971markers can be used for themarker assisted selection in breeding a new inbred line, and provide the foundation for gene isolation by mapbased cloning. 112
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京S 1.03 - Genetic Analysis of Quantitative Traits on Hybrid F1、F2 of Elymus canadensis × Elymus sibiricusLi JH1 Yun JF*,21 College of Life Science and Technology, Inner Mongolia Normal University, Huhhot 010022,P.R.of China; 2College of Ecol. And Env.Sci., Inner Mongolia Agric.Univ., Huhhot,Inner Mongolia 010019,P.R.of China.* Email:lijinghuan0816@126.com;Genetic Analysis of Quantitative Traits on Distant Hybrid F1、F2 of Elymus canadensis × Elymus sibiricuswere investigated by QTModel-Win-0.8.4Beta .And heterosis of hybrid F1,F2 was predicted. The results showedthat plant height,grass production,tiller number,Leaf Characters,Stem diameter,node number and part ofspike characters of parents , F1 and F2 were different in different environment.All this indicated that thegenotype and environment was interacted. Plant height was controlled by the interaction effect environment xadditiveand environmen x dominance, especially the interaction effect environmen x dominance. Leaf length,leaf width , leaf area, tiller number, Stem diameter,node number and spikelet number per spike were controlledby additive effect. grass production and spike length were controlled by dominant effect .Thus plant height,grass production,tiller number and floret number per spikelet should be studied in different environment.Narrow heritability of tiller number, node number, Stem diameter, spikelet number per spike and floret numberper spikelet was 51%,62%,44%,96%,99% respectively.And character improvement of this five character shouldbe done in early generation. All these results might provide evidence for Distant Hybridization Breeding in E.canadensis × E. sibiricus.S 1.04 - Genetic Improvement of Jatropha Curcas L. in HainanWen MF, Fu YH, Wang HY, Lu C and Wang WQ *Institute of Tropical Bioscience & Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou571101, China.* Email: wquanw@hainan.netJatropha Curcas L originated in Central and South America is a perennial shrub or tree in the Euphorbiaceaefamily. Jatropha widely distributed in tropical and subtropical region of the world is known as a huge potentialbio-diesel source for its oil enriched seeds (over 40%) and tolerance to drought and barren soil. However,presently its seed productivity is far too low to be commercialized because of lacking high yield and high oilcontent varieties. The real bottleneck is less knowledge about genetic resources and background in genetic andgenomics.In the study, 154 Jatropha accessions were collected from south China and other eight countries in South EastAsia, South America and Africa and conserved in Hainan. A set of 241 SSR markers cover 187 EST-SSRs and54 genomic SSRs has been developed based on transferability from near relative species cassava. Geneticdiversity evaluation of all collections by part of SSR markers shown that the population genetic diversity index Ireach to 0.55 means higher genetic diversity being, and part of domestication has took place in Yunnan andHainan population. A BIBAC genome library was constructed first timely in Jatropha, the library contains28293 clones and inserts with an average size of 131.9 kb. Meanwhile, nine genes involved in the fatty acidmetabolism have been identified from the BIBAC library. Field economic trait evaluation shown seeds oilcontent varied from 15.0% to 44.8%, and more than 400 plant lines have been selected by systemic breeding 113
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京and hybridization, part of elite lines inter yielding trail test. A molecular genetic map has been developingaccording to an experimental population with 450 lines. All these progress will help to genetic improvement ofJatropha in the future.S 1.05 - Evaluating a genepool of new type Brassica napus for its potential in intersubgenomic heterosisbreeding of rapeseedWu C, Xiao Y, Zou J and Meng JL *National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan 430070,China.* Email: jmeng@mail.hzau.edu.cnA substantial ArArCcCc genepool of new type Brassica napus has been established by combination of Arsubgenome from 135 cultivars of B. rapa and Cc subgenome from 72 cultivars of B. carinata (Xiao et al., 2010).Lines from the genepool were selected to test their combinability with traditional cultivars of B. napus(AnAnCnCn). Forty F5 lines of ArArCcCc new type B. napus were cross-pollinated with two traditional cultivars,Tapidor (from Europe) and Zhongshuang 9 (a Chinese elite cultivar), resulting 80 intersubgenomic hybrids. Thehybrids were grown in Wuhan with three replications and 225m2 for each plot. Strong intersubgenomic heterosiswas observed from the hybrid plants. Forty percent of intersubgenomic hybrids exceeded the elite commercialhybrid (control) for the seed yield, and seven intersubgenomic hybrids surpassed the control for the seed yield atthe significant level (p=0.01). The results demonstrated that the genepool of new type B. napus has a greatpotential for hybrid breeding in rapeseed.S 1.06 - An intergeneric crossing of Bambusa multiplex × Dendrocalamus latiflorus and hybrididentification by morphological and molecular analysisYuan JL*, Yue JJ, Ma NX and Chen YTResearch Institute of Subtropical Forestry, Chinese Academy of Forestry, Fuyang 311400, Zhejiang, China.*E-mail: yjl_369@sina.comBambusa multiplex, a caespitose bamboo species with wide distribution in Asia, Africa, Australasia, southAmerica, etc, is evidently popular in China on account of its relatively cold-resistance, especially in Chinesetemperate regain where other sympodial bamboo resources were scarce for winter killing, which shares goodfibre for paper making simultaneously.Dendrocalamus latiflorus, the other sympodial bamboo species with giant culms up to 25 m, is a both timberand shoot-used bamboo species which grows in Asia-temperate and Asia-tropical, however, its cultivation wasrestricted in southern China by an obstacle of winter killing.Intergeneric crossing of B.multiplex × D.latiflorus was conducted with an aim to combine the cold-hardiness ofB.multiplex and high yield of D.latiflorus. Selfing of B.multiplex and D.latiflorus were developedsimultaneously for a morphological contrast of hybrids since the difference of infants and adults, and theparental death after flowering and fruit-bearing.10 putative hybrids were selected at four months’ age by a phenotypic contrasting to paternal selfed progenies,which present an evidently large leaf than that of the maternal selfed progenies. 10 putative hybrids demonstrate 114
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京an intermediate trait between the parental selfed progenies in leaf length, leaf width, diameter, leaf veins, leafsheath, leaf auricle, etc.AFLP molecular markers were used to confirm the hybridity. 8 pairs of primer combinations were selected forgenetic analysis of parental and 10 putative hybrids samples. All of those 10 putative hybrids consist paternalpeculiar loci, with an range of 21.97%-28.10%; Maternal peculiar loci account for 15.92%-18.70%;Non-parental loci emerge with 1.88%-4.05%.Those hybrids exhibit vigorous growth performance currently in field trial and give a maximum 3.3 cm diameterand 4.0 m height, 33 cm leaf length and 6.0 cm leaf width at fifteen month’s age, which made it possible fornext elite selection after field trial. More crossing should be conducted for genetic improvement and geneticanalysis.S 1.07 - Increasing lysine content of waxy maize through introgression of o2 and o16 genes into waxy lineby molecular marker-assisted selectionZhang WL 1, 2, 4 , Yang WP 2, 3, * , Wang MC 2 , Yang LQ 2 , Wang W 2 , Zeng GP 5 , Chen ZW 1, 2 , Peng Y 4 andCai YL 11 College of Agromomy and Biotechnology, Southwest University, 2 Tiansheng Road, Beibei District,Chongqing 400715, P.R. China; 2Guizhou Institute of Upland Food Crops, Guizhou Academy of AgriculturalSciences, 1 Jinnong Road, Jinnong, Xiaohe, Guiyang, Guizhou 550006, P.R. China; 3Guizhou Key Laboratoryof Agricultural Biotechnology, Guizhou Academy of Agricultural Sciences, 1 Jinnong Road, Jinnong, Xiaohe,Guiyang, Guizhou 550006, P.R. China; 4Guizhou General Seed station, Guizhou Agricultural Committee, 62Yanan Road, Guiyang, Guizhou 550001, P.R. China; 5Agriculrural College of Guizhou University, Xiahui Road,Huaxi, Guiyang, Guizhou 550025, P.R. China.* Email: ywpmaize@hotmail.comWaxy maize (Zea mays L. sinensis Kulesh), namely sticky maize, is widely used for food product, fresh foodand dish of human, and feed and forage of livestock and birds, etc.. However, lysine content of its kernel isrelatively low, not sufficient to nutritional requirement for human, livestock and birds. In order to improve thelysine level of waxy maize endosperm, a three-way cross, [(Taixi19×QCL3021) ×QCL5019], was made andfollowed by multiple back cross and selfing, using two high lysine mutant lines, i.e. opaque-2 (o2) line Taixi19and opaque-16 (o16) line QCL3021 as donor parents and one waxy (wx) mutant line QCL5019 as receiverparent. Simultaneously, foreground selection was conducted by using SSR markers of phi027 within wx locus,phi112 within o2 locus and umc1141 linked tightly to o16 locus, so as to accurately and effectively introduce o2and o16 genes into wx line. For the offspring chosen, background selection was performed by using SSRmarkers covering whole maize genome, and the progeny seeds harvested via pollination were detected for theirlysine content by using the procedure of Acid Orange-12 Dye Binding Lysine Colorimetry and for theirstickiness character by using I2-KI dyeing method and Dual Wave Length Spectrophotometry, to ensureprogenies with high lysine, stickiness and higher restoration ratio to recurrent parent genetic background. In theBC2F2 generation, 7 plants with wxwxo2o2O16_ genotype, 19 plants with wxwxo16o16O2_ genotype and 3plants with wxwxo2o2o16o16 genotype were gained. The amylopectin content of the BC2F3 seeds derived fromthe aforementioned plants with the three genotypes was an average of 96.67%, 96.87% and 96.62%, respectively,and equivalent to that of QCL5019 seed (96.84%); the lysine content of the same BC2F3 seeds averaged out to 115
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京0.555%, 0.380% and 0.616%, and increased by 75.1%, 19.9% and 94.3% compared with that of QCL5019 seed(0.317%), respectively. The seeds of 10 pedigrees with 2 genotypes of wxwxo2o2O16_ and wxwxo2o2o16o16were reserved for planting. The restoration ratio of the 10 pedigrees ranged from 93.4% to 96.3% on QCL5019background. Therefore, pyramiding wx, o2 and o16 genes in maize by molecular marker-asisted selection canincrease lysine concentration of waxy maize or improve stickiness of high lysine maize, obtaining new type ofbreeding materials with elite nutritional and edible quality. It will be of significance for meeting food nutritionneed of human, livestock and birds, and for developing maize quality breeding.S 1.08 - 稻瘟病抗性基因等位性测验与遗传学分析张扬 稻瘟病(Pyricularia grisea)是水稻生产中的最严重病害之一,严重限制了水稻持续高产稳产。选育和利用广谱持久抗瘟的水稻抗病品种(组合) 已被证明是控制这一病害最安全、经济、环保的方法。目前生产上的问题是种植的抗性品种的抗性大多数时间短, 它严重制约了抗性品种生产潜力的发挥。为此,世界各国水稻病理学和育种学家们把水稻持久抗瘟性资源( 基因) 的发掘和持久抗瘟性育种作为抗病育种研究热点。 前人通过对抗性材料的等位性测验等方法发掘抗稻瘟病新基因,并且已经定位克隆了这些新抗病基因。结合本实验室的资源,一批高抗材料和近等基因系材料,拟通过等位性测验的方法来分析高抗材料的抗性遗传背景,发掘高抗材料持久抗瘟性基因,并进行初步定位研究。为田间持久抗瘟育种,选育广谱持久抗性水稻种质资源新品种提供有价值的参考。S 2.01 - Gene Function and Modulation of DREB (dehydration-responsive element binding protein)Genes from SoybeanChen M , Xu ZS , Li LH and Ma YZ *National Key Facility for Crop Genetic Resources and Genetic Improvement (NFCRI), Key Laboratory of CropGenetics and Breeding, Ministry of Agriculture, Institute of Crop Science, Chinese Academy of AgriculturalSciences (CAAS), Beijing 100081, China;Present address: Beijing Engineering and Technical Research Center for Hybrid Wheat, Beijing Academy ofAgricultural and Forestry Science, Beijing 100097, China.* Email: mayouzhi@yahoo.com.cnThe transcription factors DREB specifically bind the dehydration- responsive element/C repeat (DRE/CRT)cis-acting element and control the expression of many stress-inducible downstream genes in plants. Byscreening of drought-induced soybean cDNA library, we isolated four cDNA for DREB homologs: GmDREB1,GmDREB2, GmDREB3 and GmDREB5. The expression of GmDREB2 was induced by drought, high-salt (200mmol/L NaCl), low temperature (4℃) stresses and ABA (200 umol/L) treatment, and GmDREB3 was inducedby low temperature, and the GmDREB5 was induced by high-salt and drought stresses. Gel mobility shift assayshowed that these four genes specifically bound to DRE/CRT element in vitro. The transcript activating assayshowed that those genes specifically activated expression of downstream genes fused with the promotercontaining three randomly repeated copies of the wild-type DRE/CRT sequence. Functional analysis indicatedthat expression of those GmDREBs enhanced tolerance to different stresses such as drought, high salt or low 116
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京temperature, suggesting that those GmDREBs are potentially useful for improving stresses tolerance of crops. Inaddition, to elucidate the modulation of DREB transcription factors in the process of translation, some genesencoding interactive proteins with these DREB proteins were isolated from soybean using yeast two hybridsystem. To date, some interactive proteins such as E2 lingase, TPR repeat containing protein were isolated fromsoybean. Their interactive activity were proven using Put-down method. Furthermore, analysis of theGmDREB3 promoter elucidated its cold-induced modulation: a promoter fragment was involved in response tocold stress and its effect was detected for 1 h after treatment, and transcriptional repressor appeared to impairthis response by binding to a cis-element in anther region 24 h after cold stress. These results are useful forelucidating regulative mechanism of these DREB proteins under various stress condition.S 2.02 - Effects of several GDSL lipases regulated by DELLA proteins on seed fatty acids in ArabidopsisChen MX, Jiang LX﹡Institute of Crop Science, College of Agriculture and Biotechnology, Zhejiang University,268 Kaixuan Road, 310029 Hangzhou, People’s Republic of China* Email: jianglx@zju.edu.cnGibberellins (GAs) act throughout the whole life cycle of plants regulating vegetative growth and reproductivedevelopment, such as floral and embryo development. DELLA proteins including RGA, GAI, RGL1, RGL2 andRGL3, form the group of negative regulators in the GA signaling pathway in Arabidopsis. Compared to thewild-type (WT), the fatty acid content of the mature seed of the penta mutant (ga1-3 rga-t2 gai-t6 rgl1-1 rgl2-1)increased significantly. Moreover, the microarray data showed that several GDSL lipase genes weredown-regulated by DELLA proteins in unopened flower buds. In the current study, we investigated the alleliceffects of several genes (At1g54790, At1g58430, At3g48460, At4g18970 and At5g45960) coding GDSL lipaseson seed fatty acids in Arabidopsis. The corresponding T-DNA insertion mutants were backcrossed with WT forthree generations. The main fatty acids of the mutant seeds were detected by Gas chromatography. The resultsindicated that the total fatty acid content of the mutants, where the gene coding a GDSL lipase was interrupted,changed significantly, and meanwhile the major fatty acid composition was also changed. On the other hand, weover-expressed these GDSL lipase genes in Arabidopsis. The results showed that the main fatty acids in T3seeds also changed greatly. Moreover, the complementation experiments gave rise to the recovery of fatty acidcontent in the mutants to the WT level. Furthermore, q-RT-PCR was used to detect the transcriptional level ofthese genes between the Penta mutant and the WT in developing seeds. A great increase of the transcription ofthese genes in the penta mutant during 12 or 14 days after pollination were observed in contrast to WT. Inaddition, we obtained nine double mutants of the five GDSL lipase genes and analyzed the major fatty acids ofall the double mutants. The result showed that the cross-talk existed among the GDSL lipase genes. Takentogether, the study suggests that the GDSL lipase genes affect the seed fatty acids and the transcriptional patternof the GDSL lipase genes were regulated by DELLA, the negative regulators on GA signaling pathway.S 2.03 - Characterization of genes for starch metabolism from full-length cDNA libraries in cassava(Manihot esculenta Crantz)Chen X, Zhang Y, Wang HY, Xia ZQ, Wang WQ *The Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences,Haikou, China, 571101. 117
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京* Email: wquanw@hainan.netA starch metabolism related full-length cDNA library has been constructed in cassava. Two representativeManihot species, Manihot esculenta, the cultivated variety-Arg7, which had high root starch content (~32%);Manihot flabilifollia, the relative ancestral species-W14, which had low root starch content (~5%). Owing to thegreat divergence in starch content between Arg7 and W14, both total RNA samples of root and leaf wereextracted to construct cDNA library respectively, and want to find out the difference in starch accumulation andphotosynthesis between these two species. Totally, we got more than 32 000 recombinant clones, and the lengthof insert cDNA was evaluated by random samples and it possibly ranged from 300bp to 2kb. Five thousand andsixty-six 5066 recombinant clones were randomly chose to sequence and finally 5044 EST sequences were gotwhen removing low-quality sequence. All 5044 sequences were assembled into 476 contigs and 782 singletonsby using the soft of phrap and the average length of 1258 unigenes was 563.20bp. Among these 1258 unigenes,934 unigenes were annotated, and most of the annotated unigenes were located in 20 metabolism process classes,such as translation, ribosomal structure and biogenesis, energy production and conversion, carbohydratetransport and metabolism, and so on. And there are many unigenes involved in specific metabolism pathway,including glycolysis/gluconeogenesis, photosynthesis, starch and sucrose metabolism, oxidativephosphorylation, flavonoid metabolism, phosphatidylinositol signal system and ubiquitin mediated proteolysis,etc. Furthermore, part of unigenes that involves in important metabolism showed differential expressed betweenARG7 and W14 basing on their unigene size, and these unigenes can be considered as the key candidate genesto elaborate the mechanism of differential efficiency of starch accumulation in ARG7 and W14 root.S 2.04 - Cloning of PpCBF1 Gene from Peach Fruit and its Expression Characteristics during Ripeningand under Different Cold Storage ConditionsChen X, Wang GX*, Liang LS, Ma QH, Dang JLResearch Institute of Forestry,Chinese Academy of Forestry,beijing100091;2.Key Laboratory of Tree Breedingand Cultivation,State Forestry Administration,Beijing100091.*E-mail: wanggx0114@126.comDuring post-harvested cold storage of Peach fruit, chilling injury usually occurred, which made a serious declinein the quality of peach fruit and caused great economic loss; CBF play an important role in cold hardiness, butlittle is known about its roles in fruit. In this work, according to analysis ESTree database, a full-length cDNAsequence of C-repeat/dehydration-responsive element binding factor (CBF/DREB) gene was obtained frompeach fruit ( Prunus persica cv. ‘okubo’) using eletronic-PCR clone and RT-PCR, named PpCBF1 (GenbankAccession No. HM228387). Sequence analysis revealed that PpCBF1 is 776 bp in full length and contains a5-untranslated region (5-UTR) of 46 bp, a 3-UTR of 37 bp, and an opening reading frame (ORF) of 693 bpencoding a 330 predicted amino acids residues which possessed conserved AP2/EREBP domain for the CBFfunction. Sequence alignment and phylogenetic analysis revealed that PpCBF1 shared more than 90% identitywith CBF homologues from other Rosaceae plants.In addition, PpCBF1 expression profiles during fruit ripening (mature green fruit (0 days after harvest, MF),partially ripen (2 days after harvest, PR), middle-ripen (3 days after harvest, MR), fully ripen (5 days afterharvest, FR)) were analyzed by semi-quantitative RT-PCR method. With fruit ripening development, ethylene 118
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京production enhanced, simultaneity PpCBF1 transcript level was up-regulated, and this revealed that PpCBF1may be involved in ripening process. And four different treated peach fruit including ambienttemperature-storaged control fruits, 0℃storaged for 60d, 5℃ storaged for 7d, and 8℃storaged for 60d and soon were also used to study peach transcript expression patterns under cold storage conditions, the result will beanalyzed in combination with cold-chilling incidence findings index of peach fruit. Compared to ambienttemperature-storaged control peach fruit, cold-storage treatment caused increased levels of PpCBF1 expressionin peach fruit, and PpCBF1 expression abundance of 5 ℃ storaged for 7d is highest, the minor was 0℃storagedfor 60d,only little expression of 8℃ storaged for 60d was detected;The results demonstrated that PpCBF1expression showed low temperature-induced behavior in peach fruit. On the other hand, we concluded thatPpCBF1 expression was consistent to cold chilling index; PpCBF1 can be used as a transcriptional marker ofcold stress in ripening fruits of peach. We concluded that PpCBF1 may involved in cold acclimation andripening course of peach fruit. This work will improve our understanding on fundamental mechanism of peachmetabolism barrier under cold storage,on the other hand,results will lay the foundation for finding ameasurable method to maintain better quality during cold storage and carrying out genetic improvement ofchill-tolerant peach cultivar.S 2.05 - Molecular cloning, sequence and expression analyses of transcription factor ZmC4HC3 in maizeCui QX, Li YL*, Wang QL, Zhou YGCollege of Agriculture, Henan Agricultural University, 95 Wenhua Rd, Zhengzhou, China.* Email: yuling_li@126.comEndosperm accounts for more than 80% of grain weight in maize. The development, proliferation and filling ofendosperm cell play determined role in the formation of grain weight and quality. Although many studies aboutkernel endosperm have been conducted, most of them were focused on the features of starch and the roles ofkey enzymes in the process of starch synthesis. Little is known about the development mechanism of grainendosperm in different periods, especially in the molecular level. In our previous study two typical inbred linesfor grain size, a popcorn inbred N04 with very small grain and a normal dent inbred Dan232 with large grain,were chosen to reveal the differentially expressed genes in the key periods of endosperm development. FourcDNA libraries were constructed by suppression subtractive hybridization (SSH) method, using 10 d and 20 dafter pollination (DAP) endosperms of the two inbreds. In this study an EST (PE12C5), highly homologous toC4HC3 RING-type finger protein of rice, was chosen as a query probe for in silico cloning. To verify the resultof in silico cloning, specific primers were designed for RT-PCR amplification and a 1504 bp cDNA fragmentwas obtained from 10 DAP endosperm of inbred N04. This fragment was fully sequenced and considered as anew maize C4HC3 RING-type finger protein (GenBank accession no. GQ131520.1). The full-length cDNA,termed ZmC4HC3, was consisted of a 185 bp 5’-untranslated region (UTR), a 377 bp 3’UTR, and a 942 bp ORFwhich encoding for a putative protein of 314 amino acids. The deduced protein had a C4HC3 RINGv domainand 95%, 85% and 70% similarities with those in sorghum, rice and Arabidopsis, respectively. By performing ablast search against B73 RefGen_v1, ZmC4HC3 was located at bin 1.10. Fluorescent quantitative RT-PCRshowed that ZmC4HC3 expressed differently among inbreds, tissues and developmental stages. In most cases, itexpressed higher in inbred Dan232 than in inbred N04, especially for 3 DAP grain. For endosperm in 10, 15, 20,30 and 40 DAP, it expressed highest in 15 DAP for both inbreds. Also, its expression showed no tissue-specific, 119
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京with simultaneous expression in pericarp, embryo, root, stem and leaf. In 10 DAP, it expressed much higher inleaf and pericarp than in other tissues for both inbreds. But the tendency was not the same for inbred N04 in 20DAP, in which it expressed much higher in stem. Bioinformatic analysis showed that ZmC4HC3 encoded aC4HC3 RING-type finger protein. GW2, a QTL controlling rice grain width and weight, encoded a previouslyunknown RING-type protein with E3 ubiquitin ligase activity. Therefore, ZmC4HC3 might play role incontrolling gain weight in maize and could be studied in further researches.S 2.06 - Functional Variation of PSY1 for Total Carotenoids and Haplotype Effects with lcyε and CrtR-B1on the Final Biofortification of Provitamin AFu ZYTo realize the maximum improvement of provitamin A (ProVA), reducing the transformation of β-carotene todownstream carotenoids based on enlargement of the flux to the carotenoid pathway and the zeaxanthin branchshould be the most efficient way. We studied the function of phytoene synthase 1 (PSY1) gene, which encodesthe first rate-limiting enzyme in maize carotenoid biosynthetic pathway, on total carotenoids and ProVAcomponents using association mapping, as well as QTL mapping, expression and haplotype analysis. Thepresence of functional polymorphism on PSY1 was verified by linkage mapping. The functional sites for totalcarotenoid on PSY1, InDel1 upstream of the transcript start site and SNP7 on the fifth exon, were validated byassociation mapping using temperate and tropic/sub-tropic yellow maize germplasm. Uncovered cis- or trans-factors closely linked with functional site, SNP7, should exist, because it resulted in 3.3-fold expression changesbetween B73 and BY804. Haplotypes of SNP7 on PSY1, SNP216 on lcyε, and 3’TE on CrtR-B1 greatlyincreased the contribution proportion to phenotypic variation of the five major components, total, and ProVA.Additionally, lcyε and CrtR-B1 would be the prime genes for ProVA biofortification in tropic/sub-tropicgermplasm, PSY1 also should be considered when improving temperate germplasm. The convergence offavorable alleles of PSY1 from tropic/sub-tropic germplasm, lcyε and CrtR-B1 from temperate germplasmwould be the final solution to biofortify ProVA in maize.S 2.07 - Molecular cloning of the genes regulating abiotic stress tolerance from wild barley (Hordeumspontaneum C. Koch) genetic resource in Tibetan PlateauGuo WL1, Tu SW1, Wu LL1, Jiang YX1, Cong YX1, Zhang GP1, Jiang LX1, 21 Institute of Crop Science, Zhejiang University, 268 Kaixuan Road, 310029 Hangzhou, People’s Republic ofChina.2 E-mail: jianglx@zju.edu.cnAbiotic stress is a critical factor limiting crop productivity, and the improvement of crop stress tolerance is anurgent task. Our previous physiological studies demonstrated that wild barley (Hordeum spontaneum C. Koch)was more tolerant to stresses, such as salt, drought, acid, heavy metal, etc. compared to cultivated barleycultvars (Hordeum vulgare Linn.), And other investigations also indicated that wild barley adapted in harshenvironmental conditions of Tibetan Plateau and acquired stress tolerance with divergent anti-stress mechanisms.The wild barley genetic resource is therefore valuable for crop improvement. In this study, several genes relatingto abiotic stress tolerance were cloned from wild barley and their functions were analyzed. Referring to the 120
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京genomic sequences and/or ESTs of the orthologs in Arabidopsis, wheat, sorghum, rice, cultivated barley andmaize, we designed degenerated primers to isolated the alleles of 12 stress-tolerant loci, viz. HsNAH2,HsOSISP8, HsCBL2, HsCBL3, HsCLB6, HsCBL8, HsHAL3, HsHKT1, HsHKT4, by using circular first-strandcDNA-mediated RACE (cRACE). Our study showed that the DNA sequences of the loci like HsNAH2,HsOSISP8, HsCBL2, HsHKT1 are not different from the cDNAs or ESTs of cultivated barley (H. vulgare Linn.,data from web http://www.shigen.nig.ac.jp/barley/). However, the DNA sequences of other 5 loci were differentfrom their respective cultivated barley alleles. The locus HsCBL6 changed with one amino acid (S to G at 29sites) in sequence, whereas the rest loci changed with more amino acids. It might be that some abiotic stresstolerant genes of cultivated barley had changed functions in the process of artificial domestication. The allelesof these loci in wild species could be employed to improve rice cultivars by genetic engineering. Moreover, weisolated the 5-end flanking sequences of HsHAL3, HsHKT1, HsHKT4, by inverse PCR (IPCR) and nested PCR,and are investigating the capacity of the regulating sequences in model crop Arabidopsis as well as in ricecultivars.S 2.08 - Effect of the Expression of Transcription Factor Gene Mfhb-1 on the Content of 10 Kinds ofMental Element in the Somatic Embryogenesis in Alfalfa (Medicago falcata L.)He NN, Wei QC, Xue XF, Zhang SL, Zhou Y*Department of Biotechnology, School of Life Science and Technology, Henan Institute of Science andTechnology, Xinxiang, 453003, Henan, ChinaEmail:yanzhou@hist.edu.cnSubtractive cDNA cloning was used to isolate genes expressed after the early induction stage of direct somaticembryogenesis from alfalfa leaves. Some 100 different clones were identified by sequence homology andcategorized by function. The largest group of genes encoded a range of proteins involved in ribosomebiosynthesis, protein synthesis and post-translational modification. Other groups of genes coded forstress-related proteins, cytoskeletal components and regulatory proteins. One of the regulatory genes found inthis study (Mfhb-1) encodes an orthologue of the Arabidopsis HD-Zip transcription factor, Athb-1. In order tocharacterise the role of Mfhb-1 in somatic embryogenesis, constructs containing regions of the HD-Zip geneMfhb-1 in sense and antisense orientation, under the control of the CaMV35S promoter, were introduced intoalfalfa tissue by Agrobacterium-mediated transformation procedures. Transformants were successfullyregenerated. Relevant studies have indicated that the Mfhb-1 gene closely associated to somatic embryogenesisin alfalfa.In this study, young leaf cell from sense and anti-sense transgenic plants with Mfhb-1 gene was inducted by2.4-D to process the somatic embryogenesis, 10 kinds of metal elements were surveyed by inductively coupledplasma emission spectroscopy (ICP-AES) technology. The results showed that the expression of Mfhb-1 genemost likely promotes the increase of the content of Ca and Na elements while inhibits the content of Mg and Znelement during the somatic embryogenesis in alfalfa.; alternatively, the expression of Mfhb-1 had not showed anobvious impact on the change of the content of Cu, K, Fe and Mn, respectively. Further study is required toillustrate the relationship between the expression of the Mfhb-1 gene and the content of mental elements so thatto take a step leading to reveal the insight of the detailed molecular mechanism of the Mfhb-1 gene. 121
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京S 2.09 - Preliminary Study on Nucleic Male Sterile Gene by RAPD in Brassica napus L.Huang XQ1*, Hang T1,Hou GZ 2, Li L1,Hou Y2, Peng HY11 Guizhou institute of Biological Technology,Guizhou Key Laboratory of Biological Technology,GuizhouGuiyang Jinzhu, 550006,China,2 Guizhou Institute of Rape,Guizhou Guiyang Jinyang 550008,China* Email: huangxq2000@yahoo.comIn order to investigate the genetics backgrounds for Guizhou genic male sterile (GMS) rapeseed, two GMS lines(Brassica napus L.), “827AB” and "Youyan 10" were used as experiment material in this research. The latterwas bred by Guizhou Institute of Rapeseed in Guizhou Province and is widely cultivated in the upper of YangtzeRiver. Bulked segregation analysis (BSA) strategy was employed to identify the Ms gene by random amplifiedpolymorphic DNA (RAPD) markers.A total of 78 arbitrary 10-mar oligonucleotide primers were screened onthe DNA of fertile and sterile bulks, and the 16 polymorphic primers were found. DNA from individual plantsof each bulks were then used as templates for screening with these primers. The results showed that the primerBA2084 gave repeatable polymorphism between the two bulks and the individual plants in “Youyan 10” , whilethe primer LC02 did between the two bulks and the individual plants in “827AB”.The polymorphic bandgenerated by the primer LC02 was verified in the population F2 and the forecast accuracy was of 95%. Thepolymorphic bands generated by the primer BA2084 and by the primer LC02 were confirmed linkage to the Msgene of GMS.By sequencing and comparison in the GeneBank, the results showed that “Youyan 10” clip withthe primer BA2084 is partly similar to a sequence on the chromosome 1 in arabidopsis (Arabidopsis thaliana L.)at the similarity rate of 89%, and "827AB” clip with the primer LC02 is partly similar to the chromosomesegments of cabbage (Brassica rapa subsp.pekinensis) at the similarity rate of 79%.S 2.10 - Identification of Ethylene-responsive Genes in Ethrel Treated Stem Tips of Cucumber bySuppression Subtractive HybridizationJin XX, Qin ZW*, Zhou XY, Wu TDepartment of Horticulture, University of Northeast Agriculture, Harbin, 150030, China* Email: qzw303@126.comEthylene was one of the most important hormones for flowering of cucumber (Cucumis sativus L.). Themolecular interaction between cucumber and ethylene was investigated by construction and characterization of asuppression subtractive hybridization cDNA library, which was constructed from stem apex of cucumber treatedby ethrel. To screen the differentially expressed genes, dot blotting was used with cDNA probes prepared frommRNAs of control and ethrel-treated samples. Acquired positive clones were subjected to sequence andhomology search analysis.In total, 103 unique ESTs were identified from 458 positive. Among these ESTs, 73 ESTs showed significantsequence similarity to known genes, 26 had unknown functions, 4 clones did not match any known sequences.Functional categories showed that these ESTs had a wide range of roles in different biological processes. Mostof the ESTs with known functions were involved in photosynthesis, metabolism and transcriptional regulation.The complexity of cucumber in response to ethrel-treated suggested that a large number of genes were involvedin ethrel response pathways. Our study provided an overview of gene expression in ethrel-treated cucumber, as 122
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京well as useful information of the sex differentiation in plant to ethrel. Further characterization of the precisefunctions of these unique ESTs during ethrel-treated by reverse genetics will greatly contribute ourunderstanding the fundamental mechanism of ethylene response for sex differentiation of cucumber.S 2.11 - Identification of SUB1A alleles from wild rice Oryza rufipogonLi ZX1, Septiningsih EM2, Quilloy-Mercado SM2, McNally KL2, Mackill DJ21 Institute of Crop Genetics and Breeding, Yangtze University, Jingzhou City, Hubei Province, P.R.China.2 International Rice Research Institute, DAPO Box 7777, Metro Manila, PhilippinesSubmergence stress is a major abiotic stresses in the rainfed lowlands in South and Southeast Asia. Most rice(Oryza sativa) cultivars are intolerant to flooding and die within a week of complete submergence, while a smallnumber of accessions are submergence-tolerant for up to two weeks or more. These cultivars have the tolerantallele of the SUB1A gene, one of three ERF genes at this locus on rice chromosome 9. In all O. sativa varieties,the SUB1A gene is limited to a subset of indica accessions of O. sativa. Thus far, there has been no report of theSUB1A gene in wild rice species. Here we report the first evidence of the SUB1A gene found in wild species ofO. rufipogon accessions by the use of degenerate primers corresponding to the most highly conserved regions ofthe SUB1 locus. The results indicated that two SUB1A-like alleles, e.g. OrSub1A-1 and OrSub1A-2, wereidentified from two O. rufipogon accessions. Submergence treatment shows that both of the accessions withSUB1A-like genes were submergence-intolerant. This preliminary study provides insight into the origin andallelic variation of SUB1A, an agronomically important gene that is rapidly being introduced into widely-grownrice cultivars.S 2.12 - Cloning and analyzing of pathogenesis-related Gene (SiSGT1) in Setaria italica BeauvLi ZY, Dong ZP, Weng QY, Dong L, Ma JF, Quan JZ,Millet Institute,Hebei Academy of Agricultural and Forestry Sciences, Shijiazhuang 050031Leaf rust is one of the most widespread and regularly occurring diseases on foxtail millet which restrict theplanting of foxtail millet. Shilixiang is the only resistant variety from 16 000 germplasm collections beinginoculated with Uromyces setariae-italicae, but their agricultural property is very poor. Cloning rust resistantgene is helpful for breeding rust resistant foxtail millet cultivars.cDNA library of Shilixiang was constructed by SSH(suppression subtractive hybridization) method afterinoculation with Uromyces setariae-italicae isolate 93-5. Eleven sequences related to resistant gene of Setariaitalica Beauv against leaf rust infection were obtained. One sequence had 91% to 97% homology with SGT1 ofSorghum bicolor, Zea mays, Hordeum vulgare and Thinopyrum intermedium and it was namedSiSGT1(setariae-italicae SGT1, GenBank No. GU930355). The full-length cDNA sequence of SiSGT gene wasobtained by RACE amplification and its promoter region was cloned by Genome Walking. The SiSGT1 encoded359 amino acid residues and InterProScan analysis predicted that SiSGT1 included all typical SGT1 domains ofTPR, VR1, VR2, CS and SGS.The promoters of SiSGT1 were analyzed by the software of Plantcare. It indicates that the basic core elements ofTATA-box and CAAT-box, as well as stress-induced elements such as TC-rich repeats. The promoters of SiSGT1also contained cis-motif conferring response to jasmonic acid and salicylic acid such as CGTCA-motif, 123
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京TGACG-motif, and TGA-element, which suggest the gene may be induced by jasmonic acid and salicylic acidSouthern blot indicated that there was three copy of the SiSGT1 gene in the genome of Setaria italica. TheSiSGT1 gene was induced after being inoculating with Uromyces setariae-italicae by Semi-quantitative PCRmethod.The results showed that SiSGT1 had relation to resistance response of Foxtail millet against Uromycessetariae-italicae infection.Next step we will identify the function of the gene by VIGS andAgrobacterium-mediated tansformation.S 2.13 - Climatic change with bio-breedingLiu LJ, Tang XF, Yang Z, Wei LSoybean Research Institute, Heilongjiang Academy of Agricultural Sciences.* Email: nkyssbd@126.comIn recent years, climatic showed anomalies around the world. Especially in China, extreme weather such asdroughr, chilling injury, heavy rain and high temperature has occurred frequently. By 2010, in the first fewmonths, drought in southwest of China makes a huge area crops aberrant. During the same time, the north partstayed at a low temperature, flooded in Xinjiang, and a storm in northwest. All these have an big effect on ouragriculture production. The way used in conventional breeding has a lot of limites in raiseing the resist of plantsto the adversity. Molecule breeding overcomes the feature of conventional breeding, weak specificity.Additional it can improve targeted the genetic background of plant in molecule angle.Heilongjiang province locates in the north part of China, the most environmental stresses are drought, soilsalinization and chilling damage. Based on these stresses, we did some research with few soybean cultivarswhich were planted widely in Heilongjiang province. We tansfered seperatedly several relevant genes to theresistance of the adversity. These genes included 1) the hsf8 gene which heat-resistant related, 2) the bows genewater stress related, 3) the brcs gene chilling and salty resistant related, 4)the ipt gene crop production related, 5)the GmGT-2A gene and cdf4 gene which are related with water stress and the resistant to chilly and salt, and 6)the badh gene, cmo gene, GmNHX1 gene, and GmCLC1 gene which are related to the salt tolerance. And wealso constructed the dual vector which has hsf8 gene and brcs gene.Until now, we already got an array of tranfered plants. 1) The resisted plants which tansfered the hsf8 gene aretesting the T6. From these plants, we get 14 lines of early maturation variety (105 growing days), 25 lines ofmiddle maturation variety (116 growing days), and 43 lines of late variety (126 growing days). Drought stressexperiment certified these strains have improved the drought resistant and ripening enduring ability. We didsome crosses from the hsf8 transferred soybean and major soybean cultivars in Heilongjiang provience, and gota few germplasm which are under the stage of generation acceleration culture. 2) The resisted lines whichtransferred with the Bows gene, brcs gene and cbf4 gene stepped into the testing of T2. 3) The hsf8 and brcstransgenic soybean. 4) The soybean, the badh gene, cmo gene, GmGt-2A gene, GmNHX1 gene and GmClC1gene transferred seperatedly, are working on the resistant testing. 4) The ipt transgenic soybean has got stablepopulation. Steady material of the ipt transgenic has the control on the growth of negative gravity of the mainstem, increasing the node umber of the main stem, improving the growth of lateral bud, changing the apicaldominance, and changing the relation of resource and sink to increase the ratio. The outstanding expressions arethe winding number, the chlorophyll content and the pod production of semi-determinatevarieties increased,advanced the cure stage 5-7 days. At the same time, the ipt gene can stimulate the plant protein compounded and 124
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京the polysome compounded and activated, so it can increase the horizontal resistance. In resistance identificationof virus disease and Grapholitha inopinata Heinrich, incidence decreased dramatically. These showed thattransfer breeding lays the foundation of moculure designed breeding, and has the vital significance on changingthe resistant of current varieties.S 2.14 - Cloning of protein phosphatase 2A regulatory subunit gene and its expression patterns incommon wheat under abiotic stressesLiu SH1,2, MaoXG1, Liu HM2, Li A1, Jing RL1*1 National Key Facility for Crop Gene Resources and Genetic Improvement/Key Laboratory of CropGermplasm Resources and Utilization, Ministry of Agriculture/Institute of Crop Science, Chinese Academy ofAgricultural Sciences, Beijing 100081;2 College of Bioengineering, Shanxi University, Taiyuan 030006.* Email: Jingrl@caas.net.cnProtein phosphatase 2A (PP2A) is one of the major serine/threonine protein phosphatases and plays a variety ofregulatory roles in metabolism and signal transduction in plants. The PP2A holoenzyme is a composition of acatalytic subunit C, a scaffolding subunit A and various forms of regulatory subunit B. Regulatory subunit Bfamily consists of four families: B or PR55, B or PR61/56, B or PR72, B or PR93/PR110 subunit family. Theexpression levels of different B type regulatory subunits are highly diverse depending upon cell types andtissues. Therefore, the regulatory subunits determine the substrate specificity and functions of PP2A. However,their functions in abiotic stress response are largely unclear. In the present study, a novel gene, TaBβ-1, wascloned from common wheat Hanxuan 10 by in silico cloning and RT-PCR, which encodes a regulatory subunitof PP2A. Real-time quantitative PCR was used to examine TaBβ-1 expression patterns responses to differentabiotic stresses. The results showed that TaBβ-1 was up-regulated by drought, salinity, cold stresses andexogenous ABA in wheat seeding. Transgenic Arabidopsis plants overexpressing TaBβ-1 improved tolerances tosalt, freezing and drought stresses. These results indicate that PP2A regulatory subunit TaBβ-1 plays a role inplant responding to drought and osmotic stresses, and its expression levels could influence the plant tolerance tosuch stresses.S 2.15 - Towards map-based cloning of a dominant genic male sterility gene in rapeseed (Brassica napus)Lu W, Xin Q, Han X, Wan LL, Hong DF, Yang GSNational Key Laboratory of Crop Genetic Improvement, National Center of Rapeseed Improvement (WuhanBranch), Huazhong Agricultural University, Wuhan 430070.*E-mail: dfhong@mail.hzau.edu.cnRs1046AB is derived from a spontaneous dominant genic male sterility mutant Yi3A, which provides apromising alternative to the CMS system for hybrid breeding in Brassica napus. The male sterility ofRs1046AB has been proved to be control by the Ms locus which includes three alleles. However, it remainsunknown that how the multiple alleles condition the development of male gametophyte, and whether the Msgene also causes the arrest of female gametogenesis. In this study, we attempted to isolate the Ms gene by amap-based cloning strategy, in combination with the integration of different genetic mapping results and 125
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京comparative genomics between Arabidopsis and Brassica napus. Presently, the Ms gene has been restricted in asingle Brassica napus BAC clone (from a wild type canola cultivar), in which the nearest flanking markers hasdelimitated it in a physical region of 20 kb. Moreover, we also identified a BAC clone contig homologous to thetarget BAC clone. Sequence analysis of the revealed that the two homologous regions in Brassica napus genomeshare very high identity, and they show a better conserved microcolinearity with the Arabidopsis orthologue.However, gene deletion and insertion occur obviously in the target region, and among the five candidate genesthree ones can not find orthologues in Arabidopsis. Considering the fact that the pollen mother cells can notovercome the meiosis stage in the male sterile plants of Rs1046A, we pay much more attention to a LIMtranscript factor from all the candidate genes for its possible function in cytokinesis. Now we tried to isolate thecandidate genes fragment by PCR amplification from the DGMS line and construct complementary vectors, dueto the lack of the BAC clone library of it. These results have made a solid foundation for the final isolation ofthe Ms gene.S 2.16 - A gene of the xylem-specific expression in riceLu XC 1, Cong L1 , Yang T1 , Zhao KH1 and Tao CG11 Liaoning Academy of Agricultural Sciences, Shenyang, 110161, China.E-mail: luxiaoc@yahoo.com.cn or aabb2211@126.comXylem plays an essential role in the transport of water and nutrients and provides mechanical support forvascular plants survival. Many genes that were reported involved in xylem vessel differentiation, such as ligninbiosynthesis and programmed cell death. It was seldom reported that how the genes expressed in the xylemwere exclusively regulated to adapt environmental cues, such as drought and nutrient deficiency. Here, wereported the XS (xylem-specific) gene expressed specifically in the xylem of stem and leaf in rice, but in roots,the gene specifically expressed in the root cap, not in the root xylem. The gene origins from the wild rice (OryzaRufipongon Griff.) with clearly geography distribution among cultivars tested. The root volume of T1 RNAitransgenic plants was significantly decreased than wild type. Based on the gene expressed patterns and itspredicted bioinformatics, it was hypothesized that the gene may function in perceiving outer signals in the rootcap, then adapting with regulation of shoot growth of rice, the decreased root volume may be regulated by thefeedback of overground shoot growth, more details of the gene function is ongoing. Otherwise due to thediscovery of the gene, the gene promoter fused with GFP or GUS would be used as one specific marker ofxylem to study deeply xylem expression profiling under stressed conditions.S 2.17 - Comparative DNA methylation analysis of powdery mildew susceptible and resistantnear-isogenic lines in common wheatPan Lina, Liu Xiaoying, Li Xueping, Li Jiawei, Peng Yongkang, Wang Zhenying *College of Life Science, Tianjin Key Laboratory of Cyto-Genetical and Molecular Regulation, Tianjin NormalUniversity, Tianjin 300387, P. R. China; Dr. Zhenying Wang, College of Life Science, Tianjin Key Laboratoryof Cyto-Genetical and Molecular Regulation, Tianjin Normal University, Tianjin 300387, P. R. China.*Email: wzycell@yahoo.com.cnPowdery mildew is a wheat fungal disease, caused by Blumenia graminis (DC.) Speer f.sp. tritici, is one of the 126
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京most serious diseases of common wheat in the world. There are some physiological races of wheat Powderymildew, and they have a great variation speed, which results in a number of resistance genes lose effect.However, to culture new kinds of resistant wheat strains, filter new type of resistance genes are crucial forPowdery mildew prevention. To explore the changes of gene expression in common wheat infected by powderymildew, a fine wheat variety Jing 411 and its resistant near-isogenic lines (NILs) were used for MSAP(methylation-sensitive amplified polymorphism) analysis. 2891 and 3060 clear and reproducible bands weredetected in susceptible recurrent parent Jing 411 and its resistant NILs respectively, by using 64 pairs of EcoR I+ Hpa II /Msp I primer combinations. It was found that the total methylation rate in NILs was reduced from72.7% to 67% contrast to Jing 411, both the fully methylated at the internal C and hemi-methylated at theexternal C were reduced, from 27.5% to 25.4% and 45.2% to 41.6% respectively. Locus-specific DNAmethylation analysis has shown that extensive changes (both demethylation and hypermethylation) in MSAPtypes presented in NILs. It can be seen that there were 566 (17.7%) demethylation bands detected in NILs,while only 209 (6.5%) hypermethylation bands existed in NILs. By blastn analysis for 32 isolated fragmentswhich shown different changing patterns in NILs, we obtain 3 demethylation genes include Triticum urartuacetyl-CoA carboxylase (Acc-1) gene, Triticum turgidum cytosolic acetyl-CoA carboxylase (Acc-2) andHordeum vulgare subsp. vulgare seed imbibition protein (Sip1) gene. These results suggest that acetyl-CoAcarboxylase may take part in the physiologic responses against wheat Powdery mildew; moreover, furtherexprements need to be exploring for the Specific mechanisms. All together, our data support the idea that theDNA methylation plays an important role in the resistance response to Powdery mildew, and 3 DNAdemethylation genes were found. These finding provide a basis for further research on the resistant againstPowdery mildew by epigentic modification, and the culture of new kinds of resistant wheat race.S 2.18 - Expression of Cd stress response genes in ramieShe W1, Jie YC1,2 *, Lu YW, Xing HC1,2, Zhu SJ1, Wang L1, Jiang J11 Institute of Ramie, Hunan Agriculture University, Changsha 410128;2 Hunan Provincial Key Laboratory of Crop Germplasm Innovation and Utilization, Changsha 410128;*E-mail: ibfcjyc@vip.sina.comRamie (Boehmeria nivea) is a promising plant species for the ecorestoration of mining land. Ramie has beencultivated for over 5 thousand years by Chinese people and thus also named “China grass”. To investigated themechanism of Cd tolerant in ramie, two RNA libraries of Cd stress(100mg/kg) and control of ramie root weresequenced using solexa, and over 3.5 million short sequence reads were produced, Clean tag accounted for93.18% and 93.76% separately. 3887 differently expressed genes of ramie root under Cd stress were selectedreferring to the significance of digital gene expression profiles, including 2883 up-regulated and 1004down-regulated. By blasting search to NCBI, we found most of differently expressed genes were unknown. Theresults of gene ontology functional enrichment analysis for differently expressed genes showed that mainfunctions of these genes were binding and catalytic activity and the main cell component which these genesenriched were plasma membrane and vacuole. The main pathways of these genes were glutathione metabolismand cysteine and methionine metabolism. This laid foundation for understanding the mechanism of Cd tolerantin ramie from level of gene expression. 127
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京S 2.19 - Identification of a new blast resistant gene from Dacca6, a useful donor to improve the widespectrum resistance of Jin23 against rice blast fungi (Magnaporthe grisea) in Southeast ChinaShi BH1, Zhang JH1, Zheng YM2, Liu YQ2, Zheng TQ3,*, and Zhao MF2,*1 College of Life Sciences, Fujian Normal University, Fuzhou, Fujian 350108, China;2 Rice Research Institute, Fujian Academy of Agricultural Sciences, Fuzhou, Fujian 350018, China;3.Institute of Crop Sciences/National Key Facility for Crop Gene Resources and Genetic Improvement, ChineseAcademy of Agricultural Sciences, 12 South Zhong-Guan-Cun St., Beijing 100081, China;*Email: tzheng@caas.net.cn & zhmingfu@163.comJin23 is a major parental line being widely used in China’s three-line hybrid rice breeding. Its excellent grainquality has filled the gap between the high yield potential and good grain quality which bothering the hybridrice breeding for quite a long time. However, it was quite sensitive to rice blast fungi (Magnaporthe grisea),especially in Southeast China, such as Fujian province. In order to improve the resistance of Jin23, new geneticresources are highly required. In 2005, 130 molecular breeding parental lines imported from the global ricemolecular breeding network initiated at IRRI have been screened by natural inoculation at the blast endemicfield in Fujian. Among them, Dacca6, a local variety from the Philippines, was found to possess wide-spectrumand high resistances to the major races in Fujian. The following report focused on the further identification ofthe resistance by artificial inoculation and the mapping of the underlying genetic locus as well as its applicationin the improvement of Jin23B (maintainer) for the blast resistances.Two subsets of a BC1F2 population derived from the cross of Dacca6 with Jin23B were inoculated by sprayingwith two representative isolates, SM03023 and SH0512, respectively, at the seedling stage. Both of thesegregation results from these two subsets pointed to a same new single dominant gene from Dacca6. Bytraditional linkage mapping, this gene was tagged between markers RM5529 and RM211, the genetic distancebetween these two markers and the gene was 3.8 cM and 4.1 cM, respectively, on chromosome 2. Furthermore,we assayed the DNA pools of resistant and susceptible individuals from independent BC1F2 populations derivedfrom eight BC1F1 resistant plants with 21 SSR polymorphic makers on chromosome 2 to identify theintrogression patterns. It was found that all the pools were heterozygous at the marker locus of RM211, whichindicated that the resistance gene was closer to RM211. This was consistent with the results from the previouslinkage mapping work. With the aid of comparative mapping tools, we compared the candidate position with theknown blast resistant genes, and no reported gene was found in this region. The existence of a new resistantgene in this candidate region was strongly indicated.Based on the mapping results, after consecutive backcrossing followed by selfing with the aid of molecularmarker aided selection (MAS) by RM5529 and RM211, twelve Jin23B BC3F3 lines were obtained. Theresistances of these lines were confirmed by a field evaluation in the blast epidemic area, Jiangle County, Fujian.One of these elite lines was further evaluated for its agronomic traits, and no significant difference was found incomparing with Jin23B.This study has preliminarily showed a way to breeding blast resistant Jin23B with new gene from Dacca6 byMAS. The development of blast resistant Jin23 male sterile line is undergoing based on the lines from this work.Meanwhile, the mapping information will also help on the related gene cloning work in the near future. 128
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京S 2.20 - DNA allelic variations at the loci putatively implicated in seed oil formation among Brassicaoilseed cultivarsShi C 1, Zhu Y 1, Li Y, Guo W, Chen K, Shamsi I, Hua S, Zhong Z, Zhou W 2 and Jiang L 2Institute of Crop Science, Zhejiang University, 268 Kaixuan Road, 310029 Hangzhou, China.Email: wjzhou@zju.edu.cn; jianglx@zju.edu.cnKnowledge about the genes implicated in lipid biosynthesis acquired from the model plant Arabidopsis is usefulin understanding the formation of seed oil in Brassica oilseeds. In this paper, we report the screening ofpolymorphic markers at the loci putative for the seed oil formation between two geographically differentgenotypes: the Chinese cultivar Ningyou-7 and the European cultivar Tapidor. These primer pairs (150) weredesigned based on 75 Brassica genes that were Arabidopsis orthologues implicated in the oil formation. A totalof 52 out of the 150 primer pairs associated with 47 of the 75 genes showed polymorphisms between the twogenotypes. The type of polymorphisms that could be detected on capillary electrophoresis images and theirrespective visual futures are described. Further, we selected 34 polymorphic markers to scan allelic variationsand found rich DNA polymorphisms among the 54 Brassica oilseed cultivars. On the average, each primer pairresulted in 5.6 alleles at the region that was covered. The correlation between the alleles and seed quality traitsrevealed that the alleles of BnFAD7 were related to the variation of linolenic acid (C18:3) contents among thecultivars. The allele FAD7-ics11170 (3/4)-b that was significantly correlated with high linolenic acid content canbe used as an efficient marker for the selection of breeding materials with high linolenic acid content.S 2.21 - An Improved Method for Extraction of Rice Cytoplasmic GenomeWan WT 1,2, Dong Y3, Yu Y2 , Li LJ2, Zhang J2 , Li Q2 , Hu FY 2*1 Academy of Agriculture and Biotechnology,Yunnan Agricultural University,Kunming,650201;2 Food Crops Research Institute,Yunnan Academy of Agricultural Sciences,Kunming,650205;3 State Key Laboratory of Genetic Resources and Evolution,Kunming Institute of Zoology,Chinese Academyof Sciences,Kunming,650223* Email: hfengyiyaas@yahoo.com.cnChloroplast and mitochondria of plant have their own independent replication and inheritance genome. Due tothe small genome,how can we separate and purificate the high-quality cytoplasmic genome is the prerequisiteto research cytoplasmic effect on DNA level. In this research,we have been developing a new technique basedon combination of sucrose step-gradient centrifugation and whole genome amplification(WGA). Through thedeveloped method,we can get high quality and quantity(OD2801.8~2.0)chloroplast DNA and mitochondrialDNA with about 30ng/ul of concentration and 1.5ug of grosses from only 20g rice leaves comparing withtraditional methods. The chloroplast DNA and mitochondrial DNA can fulfill the needs of many cytoplasmicgenome study,even the needs of new generation sequencing technology.S 2.22 - TaABC1, a member of the activity of bc1 complex protein kinase family from common wheat,confers enhanced tolerance to abiotic stresses in ArabidopsisWang CX , Mao XG , Chang XP , Li Ang ang Jing RL *The National Key Facility for Crop Gene Resources and Genetic Improvement, Institute of Crop Science,Chinese Academy of Agricultural Sciences, Beijing 100081, China.*Email: jingrl@caas.net.cn 129
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Abiotic stresses such as drought, salinity and low temperature have drastic effects on plant growth anddevelopment. However, the molecular mechanisms regulating biochemical and physiological changes inresponse to stresses have not been well understood. Protein kinases are major signal transduction factors amongthe reported molecular mechanisms mediating acclimation to environmental changes. Protein kinase ABC1(Activity of bc1 complex) is involved in regulating coenzyme Q biosynthesis in mitochondria in yeast(Saccharomyces cersvisiae), and in balancing oxidative stress in chloroplasts in Arabidopsis (Arabidopsisthaliana). In the current study, TaABC1 (Triticum aestivum L. activity of bc1 complex) protein kinase waslocalized to the cell membrane, cytoplasm, and nucleus. We further investigated the effects of overexpressingTaABC1 in transgenic Arabidopsis plants on responses to drought, salt and cold stress. Transgenic Arabidopsisoverexpressing the TaABC1 protein showed lower water loss and higher osmotic potential, photochemistryefficiency and chlorophyll content, while maintained cell membrane stability and controlled reactive oxygenspecies (ROS) homeostasis. In addition, overexpression of TaABC1 increased the expression ofstress-responsive genes, such as DREB1A, DREB2A, RD29A, ABF3, KIN1, CBF1, LEA and P5CS detected byreal-time PCR analysis. Our results suggest that TaABC1 overexpression enhances drought, salt and cold stresstolerance in Arabidopsis, and imply that TaABC1 may act as a regulatory factor involved in a multiple stressresponse pathway. In addition, we developed SNP markers of TaABC1, and mapped it on chromosome 3AL,3BL and 3DL by the ITMI recombinant inbred lines (W7984 × Opata85), respectively.S 2.23 - Identification and characterization of two microRNAs and their target genes in OryzalongistaminataWang D 1, Zhang T 1,2, Hu FY 1,3, Zhu LH 1, Fu BY 1 and Li ZK 1,41 Institute of Crop Sciences/National Key Facility for Crop Gene Resources and Genetic Improvement, ChineseAcademy of Agricultural Sciences, 12 South Zhong-Guan-Cun St., Beijing 100081, China;2 College of Life Sciences, Wuhan University, 430072, China;3 Institute of Food Crops, Yunnan Academy of Agricultural Sciences, Kunming 650205, China; 4InternationalRice Research Institute, DAPO Box 7777, Metro Manila, the Philippines.In this experiment, the expressions of 90 miRNAs in five kinds of tissues (rhizome tip, rhizome internodes,shoot tip, shoot internodes, root) of Oryza longistaminata were studied. The results displayed miR221 andmiR16 expressed differently in five kinds of tissues. MiR221 and miR16 expressed more highly in rhizomeinternodes and root than in other three tissues. Via bioinformatics methods, the two miRNAs’ target genes werepredicted. Real-time PCR were used to identify the predictions. Results showed miR221 and miR16 regulate theexpression of different genes in different tissues. MiR221 regulate the expression of LOC_Os04g57610 in allfive tissues, which encode auxin response factor8. MiR16 regulate the expression of LOC_OS05g34220.2 inrhizome tip, rhizome internodes, shoot tip, shoot internodes, which encode vrga1.S 2.24 - A NCED4-like gene from Caragara korshinskii is involved in responding to variety of abioticstressWang XM, Gao HW * 130
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Inst. of Animal Science, Chinese Academy of Agricultural Science, Yuan-Ming-Yuan West Rd., Haidiandistrict, Beijing 100193, P. R. China.* Email: Gaohongwen@263.netAbscisic acid(ABA)is an essential phytohormone that plays critical roles in the regulation of physiology,growth and development in higher plants, and also is involved in plant adaptation to environmental stresses suchas salinity, drought and cold (Leung and Giraudat, 1998). In higher plant, 9-cis-epoxycarotenoid dioxygenase(NCED) cleaves C40-cis-epoxycarotenoids to produce xanthoxin, which is the key regulatory step in ABAbiosynthesis (Kende and Zeevaart, 1997).The first NCED cDNA was obtained from the ABA-deficient Vp14 mutant of maize (Tan et al., 1997). Up tonow, a number of NCED genes have been cloned and characterized in different plant species. In many species,NCED-like genes comprise a small multigene family, with different expression pattern and genomic function(Chernys and Zeevaart, 2000; Rodrigo et al., 2006).The expression of NCED genes, and the subsequent regulation of ABA biosynthesis, has been studiedextensively in several plant species. Most research focused on NCED1 gene, whereas few studies on otherhomolog, especially NCED4 gene, have been reported. In Arabidopsis, AtNCED4 gene was not induced bydrought stress.Caragara korshinskii is a deciduous perennial shrub of sandy grassland and desert. The species is distributedacross half-fixed and fixed sandy regions in northwest China and Mongolia (Fu, 1989). Although it has beendemonstrated in the literature that the species is highly stress tolerant (Zhang, 1994; Wang et al., 2007), mostresearch on C. korshinskii has focused only on the introduction and domestication of the species. Using RACEtechnology, a full-length cDNA-encoding NCED4 gene was isolated and characterized from the leaves ofCaragana korshinskii (Peashrub). The 2067-bp full-length CkNCED4 had a 1758-bp ORF, which encodes apeptide of 586 amino acids. The deduced amino acid sequence of CkNCED4 protein shared high identity withother NCED4 and shared 37.5% identification with CkNCED1, which is a NCED1-like gene and was reportedin our previous research. Southern blot analysis revealed that the gene CkNCED4 was a single copy in thegenome of C. korshinskii. CkNCED4 gene sustained abundant expression under normal condition. Quantitativereal-time PCR analysis showed that dehydration and salt stress all down-regulated the mRNA accumulation,while exogenous ABA and low-temperature stress could not affect the gene expression. The results indicatedthat CkNCED4 maybe played a negative regulation role in drought and salt tolerance in plant, at least inC.korshinskii.S 2.25 - Transform the Cre/ Lox-3 Gene into Indica Rice and Analysis of Its Storage DurabilityWei YD 1,2, Xu HB 1,2 , Zhang JF 1,2 * and Xie HA 1,2 *1. Key Laboratory of Crop Germplasm Utilization, Ministry of Agriculture, P.R.China; Fujian EngineeringLaboratory of Crop Molecular Breeding; Fujian Key Laboratory of Rice Molecular Breeding; Fuzhou Branch,National Rice Improvement Center of China, Fuzhou, 350003;2.Rice Research Institute, Fujian Academy of Agricultural Sciences, Fuzhou, 350018 ;*Email: jianfzhangrice@yahoo.com.cnRice is one of the most important grain crops in the world. Almost half of the world’s population takes it as the 131
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京main source of food and energy. Therefore the stable increasing of rice yield ensures the world’s grain supply.Furthermore, as the model plant in grass family genome research, rice plays an irreplaceable role in thebio-science research.In China, the largest rice consuming and producing country, there is more than 65% population living on it. Forover half a century, with the development of molecular biology technology, using molecular marker-assistedbreeding, genetic engineering and other related technologies, our country has achieved remarkable success inrice quality improvement and grain yield growth. However, with the large population of our country, the level ofper capita share is low, and there is also a large loss caused by aging deterioration, pests and mildew during ricestorage. Therefore, carrying out a series of related researches on rice storable characteristics and breeding newstorable rice varieties, just like improving rice production, are also very important.This study was designed to transform Cre/Lox-3 gene into indica restorer line “Hang No.2”, expecting to selectlipoxygenase low-expression lines from transgenic plants, breed new varieties with high storage resistance, andachieve the purpose of improving rice resistance to storage at last.S 2.26 - Tanscriptome profile analysis of ramieXing HC 1,3, Zhong YL 1, Zhou JL 2, Jie YC1,3 , She W 1, Zhu SJ1 and Wang L 1 Jiang J 1, Zhou JH 1 *1 Institute of Ramie,Hunan Agriculture University,Changsha 410128;2 College of Life Sciences, Hunan Normal University, Changsha,Hunan 410081, PR China;3 Hunan Provincial Key Laboratory of Crop Germplasm Innovation and Utilization, Changsha 410128;* Email: ibfcjyc@vip.sina.comRamie (Boehmeria nivea (L.) Gaud) (Urticaceae, Boehmeria) is perennial herb, originated in China. Ramie ismainly for the purpose of harvesting fiber to cultivation, but also can be used to make paper, feed, industrialalcohol and medicine. Transcriptome information represents the full transcription of genome, it was necessaryfor understanding of genomic functional elements and verifying changes of interaction among phenotype,genotype and environmental factors. The new generation sequencing technology makes researchingtranscriptome easily. For perennial features, heterozygosity and the genetic background is complex in ramie, theprogress has been slow in molecular biology of ramie, especially in functional genomics. Therefore, it couldprovide gene information for the study on molecular biological on ramie, that the transcription of all expressedgene in a certain period were obtained by solexa sequencing technology. In this paper, RNA of rootlet, bark,stem apex, leaves and bud on ramie varieties "XiangZhu 3" were extracted in earlier stage of inflorescence, thenthe transcriptome profile of ramie were obtained by solexa sequencing technology firstly. 1989040650ntnucleotides were measured and 26,530,524 reads were got, then 48,547 unigene were assembled bySOAPdenovo software. 29,689 unigenes which were blastx in nr database had similar proteins. When blastxwere done in Swiss-Prot database, 21,607 similar functional proteins were discovered. While compared unigenewith COG database, only 10,903 similar proteins were found. When compared with the KEGG database, weobtained 14,074 unigene similar proteins. And each unigene metabolic pathways were predicted in the KEGGdatabase, all the 14,074 unigenes belonging to 218 metabolic pathways. Using the best blast results to determinethe direction of the Unigene, when the Unigenes were not similarity the above four databases, we used thesoftware of ESTScan to predict the coding region and determine the direction of sequences. 32,346 unigenescould determine the orientation in this research. About 33.37% of unigenes were no orientation. GO 132
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京classification analysis was done in all functional unigenes, it could be divided into three categories, 30.76% ofthe functional unigenes were involved in biological process, 44.45% of the functional unigenes belonged tocellular component, 24.79% of the functional unigenes were participated in molecular function. It could providesupport for transcriptome research of ramie and other perennial, heterozygosity plants using solexa sequencingtechnology. It also laid the foundation of the other molecular biology study in ramie.S 2.27 - Gene expression changes in response to chilling stress in maizeYang G, Su SZ, Li SP, liu HK and Yuan YP *College of Plant Sciences, Jilin University, Changchun 130062, China.* Email: yapingyuan@hotmail.comMaize (Zea mays L.) is the crop with the largest cultivated area in the Northeast of China. Chilling stress is oneof the most important abiotic stress limiting growth, development and distribution of maize. In the present study,to understand the underlying molecular mechanism in response to low temperature, we investigated theexpression of genes in maize under low temperature using cDNA-amplified fragment length polymorphism(cDNA-AFLP). For chilling stress treatment, the plants were kept in a 4°C refrigerator for 2 h,6h and 12h,respectively. According to the cDNA-AFLP analysis, approximately 10,000 transcript-derived fragments (TDFs)were displayed using total 336 primer combinations, included 112 MseI/EcoRI (14×8) and 224 MseI/PstI(14×16) primer combinations. A total of 30-50 AFLP bands have been detected per combination onautoradiography, ranging from 70 to 600 bp. Four hundred and forty one TDFs induced by low temperaturewere obtained and 114 TDFs were successfully sequenced. Sequence alignments showed that several candidateTDFs shared high levels of identity with corresponding genes from Avena sativa, Oryza sativa, Arabidopsisthaliana, and other species. These candidate TDFs were involved in metabolism, photosynthesis and energy,transport, signal transduction, secondary metabolism and defence response, suggesting that maize undergo acomplex adaptive process in response to low temperature. The expression patterns of seven novel candidategenes were confirmed by semi-quantitative RT–PCR analysis which might associated with chilling tolerance inmaize. A significant outcome of this study is the identification of novel candidate genes that can provide newinsights and better understanding of the underlying physiological, biochemical, and molecular events involvingin maize adapting to low temperature environments. Further work will focus on the determination of the role ofthese genes in maize by transgenic approach.S 2.28 - Identification of differentially expressed miRNAs and their target genes between salt-tolerant andsalt-sensitive cotton cultivars under salt stressYin ZJ 1, Liu YD 1, Yu JW 2, Han XL 1 and Shen FF 1*1 State Key Laboratory of Crop Biology, College of Agronomy, Shandong Agricultural University, Tai’an,Shandong 271018, P.R. China;2 Cotton Research Institute, Chinese Academy of Agricultural Sciences, Anyang, Henan, 455100, P.R. China.Email: ffshen@sdau.edu.cnMicroRNAs (miRNAs) are a highly conserved class of small noncoding RNAs that regulate gene expression atthe post-transcriptional level. In plants, miRNAs play important roles not only in multiple developmental 133
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京processes but also in the response to environmental stimuli. Salt stress is one of the important elements forreduced crop production. In order to adapt to high soil salinity, plants express a variety of genes to enhance theirtolerance. However, the regulatory networks governing these genes are poorly understood. Cotton (Gossypiumhirsutum L.) is considered a relatively salt-tolerant non-halophytic species. Variations in salt tolerance have beenobserved among different cultivars. In this study, the salt-tolerant cotton cultivar (TOL cv.) SN-011 and thesalt-sensitive cotton cultivar (SEN cv.) LM-6 was used to detect and annotate differentially expressed miRNAsinvolved in salt tolerance. Using miRNA microarray analysis and a bioinfomatic approach, 12 conservedmiRNAs were identified in cotton. Among them, ghr-miR156d, ghr-miR159, ghr-miR167a, ghr-miR167b, andghr-miR399b were newly identified. Seven miRNAs were confirmed with a genotype-specific expression modelin response to high salinity. They were grouped into three categories: (1) miR156a, miR156d, miR156e, andmiR169 were dramatically down-regulated by salt stress in TOL cv., but were not affected in SEN cv.. (2)miR159 was more sustained in TOL cv., but was down-regulated in SEN cv.. (3) miR167a and miR399a weredramatically up-regulated in TOL cv., but were not affected in SEN cv.. These differentially expressed miRNAscould be of interest in determining the contrasting responses to salt stress of the two cotton genotypes. To gaininsight into their functional significance, 23 target genes were further predicted. Ghi.13781, Ghi.16537,Ghi.18144, and Ghi.10300 were targeted by miR156. They encoded squamosa-promoter binding-like proteins(SPL) which involved in vegetative phase changes. Ghi.14555, Ghi.20503, Ghi.21682, and Ghi.2038 weretargeted by miR167. They encoded Auxin response factors (ARF6/8) which can bind to auxin responsepromoter elements and mediate gene expression responses to auxin. Two miR169 targets, Ghi.713 and Ghi.9444,encoded heme activator protein (HAP2). These target genes have been verified previously. Quantification oftheir transcripts revealed that these target genes showed significant inverse correlation with correspondingmiRNAs in expression under salt stress conditions. Our results suggested that to enhance the salt tolerance ofcotton, the down-regulated miR156 and miR169 enhance the amount of SPL and HAP2 transcription factorsrespectively, further with an ultimate operation on the physiological and morphological adaptation to salt stress.The salt-induced miR167 leads to the down-regulation of ARF and acts as a responsive component of auxinsignalling pathways to enhance salt tolerance.S 2.29 - Single nucleotide polymorphism and molecular mapping of the drought tolerance geneTaSnRK2.7-B in common wheatZhang HY 1, 2, Mao XG 1 , Chang XP 1 , Xie HM 2 and Jing RL 1*1. The National Key Facility for Crop Gene Resources and Genetic Improvement, Institute of Crop Science,Chinese Academy of Agricultural Sciences, Beijing 100081, China;2. College of Agronomy, Northwest A & F University, Yangling 712100, Shaanxi, China.*Email: jingrl@caas.net.cnTaSnRK2.7, a SnRK2 (sucrose non-fermenting1-related protein kinase 2) member of wheat, confers enhancedmulti-stress tolerance in carbohydrate metabolism. Here, nucleotide diversity of TaSnRK2.7 was investigated in52 wheat accessions with different drought-resistant features. One copy, named TaSnRK2.7-B, was detected in25 accessions, which mainly exhibited drought tolerance. The TaSnRK2.7-B evolutionary dynamics wereexamined in these cultivars, which have been subjected to population bottlenecks and intensive selection inbreeding. 134
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Nucleotide diversity (π) in the entire TaSnRK2.7-B region was 0.00238, and nucleotide diversity in non-codingregions (0.00261) was higher than that in the coding region (0.00232). Sliding-window analysis demonstrated3932-4535 bp was a variation-enriched region, covering the eighth intron, the ninth exon and the 3-flankingregion. As predicted, neutrality tests revealed that TaSnRK2.7-B protein was relatively conservative, andpopulation bottlenecks or purifying selection may have acted on TaSnRK2.7-B. Furthermore, strong linkagedisequilibrium between SNP loci extends across the entire TaSnRK2.7-B region. Phylogenetic analysis showedthat two drought-sensitive and two high drought-resistant accessions were clustered in the same clades,respectively. A SNP marker of TaSnRK2.7-B was developed and mapped it on chromosome 2AL flanked byWmc179.4 and Wmc401, which were co-located in the same or adjacent chromosome intervals with QTLs forphosphorus utilization efficiency and accumulation efficiency of stem water-soluble carbohydrates. Our resultsindicated that TaSnRK2.7-B may be useful for carbohydrate metabolism and drought tolerance improvement inwheat.S 2.30 - Isolation and characterization of the Secale NPR1 (Sc NPR1) from a Chinese rye varietyJinzhouHeimaiZhang X, Ma HX, Xu L, Sun XB and Yu GHInstitute of Biotechnology, Jiangsu Academy of Agricultural Sciences, Nanjing, 210014, P. R. China.Email: Xuzhang@jaas.ac.cnThe development of transgenic wheat plants highly resistant to a range of pathogens using traditional signalgene expression strategies has been largely ineffective. Modification of systemic acquired resistance (SAR)through the over expression of a controlling gene such as NPR1 (non-expressor of PR genes) offers an attractivealternative for augmenting the wheat plants innate defense system. To find new gene resource for wheatpathogen-resistance genetic engineering, Secale cereale L. var. JinzhouHeimai was used as a starting material toclone full-length cDNA of NPR1 gene by homologous cloning and smarter-RACE techniques. In the ScNPR1molecule, there are four “W- boxes” in the upstream of ATG that are necessary for both induction of NPR1transcription and NPR1-mediated activation of plant defense responses. The gene contains the BTB and ankyrinrepeat domain that is known to be the molecular basis for NPR1 function in Arabidopsis. The predicted aminoacid sequence of ScNPR1 had highest sequence similarity to TaNPRI and OsNPR1 with 72% and 65% ofidentities but lower homology (37%) to the AtNPR1 proteins.There was some apparent induction of ScNPR1gene after SA treatment to JinzhouHeimai at three-leave stage.The gene expression level continued to increase in the plantlet, peaking at 8 h with thirty-fold increase, anddecreased rapidly to twofold increase by 24h. Increased ScNPR1gene expression was also observed after JAtreatment. A greater than forty-fold increase in gene expression was detected at 8 h, compared to twofoldexpression level at 24h. Similar expression level change of ScNPR1gene was detected during 24h after ethylenetreatment which indicated that ScNPR1gene was the key positive regulator both in the SA induced SAR signaltransduction pathway and in the JA/ET dependent ISR pathway.ScNPR1gene expression level periodically changed after inoculated by isolate of Blumeria graminis f. sp. Tritici.It was reported that formation of primary germ tube (PGT) and germ tube of appressorium (AM) of conidiaspores were observed at 3 hours after inoculation (hai) and 10 hai, respectively, while AM were observed at 24hai which induced papillae formed by infected cell. The increased ScNPR1gene expression occurred after the 135
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京time of PGT or AM formation which indicated that the expression of ScNPR1gene might respond to theinfection of pathogen.The modified pAHC25 plasmid, containing the ScNPR1 and bar genes as well as nos terminator under thecontrol of the maize ubiquitin promoter, was transferred into wheat by particle gun bombardment of immatureembryos. The transgenic wheat plants expressing ScNPR1gene has been generated. Resistance to fungaldiseases of Fusarium head blight, Powdery mildew and Sharp eyespot would be evaluated in T1 and T2generations.S 2.31 - Identification of Differentially Expressed Genes by cDNA-AFLP Technique during Water Stressin Wheat LeavesZhang YQ, Li JF, Wang Z, Fan ZR and Wu ZLInstitute of Nuclear and Biological Technologies, Xinjiang Academy of Agricultural Sciences.E-mail: fzr518@163.comXinjiang is one of the most arid areas in China, within which the oasis only occupies 4.27% of the total area.Drought is one of the main environment stress factors limiting spring wheat productivity in Xinjiang regions.The study on mechanism of corps drought tolerance has become one of hotspots in the field of crops sciencebecause of serious shortage of water resource worldwide. To improve crops productivity, it is worthful to findthe molecular mechanism of drought-resistance crops for crop breeding and cultivation techniques.In this study, seedlings of spring wheat cultivar Xinchun6 with drought resistance and high water-use-efficiencywere treated with water defict and full irrigation, and the differential expressed sequence tags (ESTs) with thedrought resistance and the water use efficiency were investigated using cDNA-AFLP technique. A total of 21nonredundant ESTs were idenfified.These ESTs were classified into several functional groups with Blastxanalyses and functional annotation, mainly including signal transduction, transcription regulation, energymetabolism, cell growth and development, stress responses. The details are as follows: (i) The signaltransduction-related genes in response to water stress: TDFHh3-3 was related to the TonB-dependent receptorgene; (ii) The transcriptional regulation-related genes in response to water stress: TDFHh1-3, TDFHh6-4,TDFHh7-1, and TDFHh20-2 showed higher similarity to the retrotransposon and reverse transcriptase genes; (iii)The energy metabolism-related genes in response to water stress: TDFHh2-1, TDFHh4-3 and TDFHh25-2 weresimilar to the cell wall-associated hydrolase gene and uracil-DNA glycosylase gene; TDFHh16-2 was similar tothe pyruvate kinase gene; TDFHh14-2 was similar to the Serine/threonine kinase gene; TDFHh15-2 was similarto the mitochondrial DNA in rice; TDFHh8-2, TDFHh12-3, and TDFHh26-1 were similar to the amino acidtransporter-related gene; (iv) The defense gene-related in response to water stress: TDFHh9-3 was homologousto the nuclear protein EAST gene; TDFHh13-2 was homologous to the stem rust resistance protein RPG1 gene;and TDFHh19-2 was homologous to the ischemia/reperfusion inducible protein gene; and (v) The unknownfunctional gene in response to water stress: TDFHh10-2, TDFHh17-3, TDFHh21-5, and TDFHh27-2 washomologous to the hypothetical or putative protein gene. These results provide information for furtherelucidation of the molecular mechanism of drought-resistance in wheat.S 2.32 - Cloning and functional characterization of a NBS-LRR that participates in wheat defenseresponse to BYDV infection 136
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Zhao D1,2, Li N 1, Huang X 1,2, Zhao JR 1, Du LP 1,2, Huang ZJ 2, Zhang ZY1*1 National Key Facility for Crop Gene Resources and Genetic Improvement / Institute of Crop Sciences, ChineseAcademy of Agricultural Sciences, Beijing 100081, China; 2 Biology College, Hebei Normal Unversity,ShiJiaZhuang050016, China.* Email: zhangzy@mail.caas.net.cnMany plants contain nucleotide-binding domain and leucine-rich repeat (NB-LRR)- type immune receptor thatfunction during defense against pathogens. Barley yellow dwarf virus (BYDV) causes significant yield losses ofwheat worldwide. No resistance to BYDV has been found in common wheat. Thinopyrum intermedium, a wheatwild relative, shows highly resistant to BYDV infection. The distal segment of Thinopyrum intermedium 7Ai#1chromosome long arm carries a BYDV resistance gene (Bdv2), and has been introgressed into wheat-T.intermedium translocation lines. In this study, a full-length cDNA encoding NB-LRR protein namely TNBL1was isolated from wheat-T. intermedium translocation line YW642 based on a differentially- expressingfragment identified by cDNA-AFLP analysis. The deduced protein consists of 1059 amino acid residues, andcontains a coiled-coil (CC), and a NBS domains, and LRR region. Sequence alignment and phylogeneticanalysis indicated that the protein TNBL1 is a new member of NB-LRR family. Like active-functionalresistance protein MLA in barley, subcellular localizations of TNBL1 were the cytoplasm and nucleus,evidenced by transient expression of yellow flurenscence protein fused with TNBL1 in onion epidermal cells.To investigate if the gene TNBL1 plays roles in wheat defense responses to BYDV, we generated transgenicwheat plants with the gene over-expressing or RNA interfering constructs. In the loss-of-function experiments,compared to the resistant recipient-parent, wild-type wheat alien translocation YW642, the expression levels ofTNBL1, SGT1 and RAR1 in these TNBL1 RNAi plants were reduced, importantly, the knocked-down of TNBL1transcript caused the BYDV resistance loss. In the gain-of-function results, compared to the susceptiblerecipient-parent, wild-type wheat Zhong8601, the expression levels of TNBL1 in the positive transgenic plantswere markedly increased, in turn, these transgenic plants overexpressing TNBL1 displayed resistance to BYDV.These results suggest that TNBL1 gene upstream of SGT1 and RAR1 participates positively in defense responseto BYDV infection.S 2.33 - Cloning and Sequence Analysis of Ramie β-amylase GeneZhou JH1, Xing HC1,2, Jie YC1,2 , Zhong YL1, She W1, Zhu SJ1, Jiang J1, Wang L1 *1 Institute of Ramie, Hunan Agriculture University, Changsha 410128;2 Hunan Provincial Key Laboratory of Crop Germplasm Innovation and Utilization, Changsha 410128;*E-mail: ibfcjyc@vip.sina.comThe liveness of the β-amylase are closely related on starch output and quality, there was no report that theβ-amylase gene has been cloned in ramie. In this study, on the basis of ramie transcriptome sequencing to searchβ-amylase genes, we has been obtained Unigene6733.According this sequence to design 5RACE and 3RACEspecial primers, the full length of the β-amylase gene sequence has been successfully cloned. The ramieβ-amylase gene has been compared with nucleic acid sequences in Genbank. The Ramie β-amylase Gene sharedgreater than 90% homology with Arabidopsis thaliana、Ricinus and potato, and shared greater than 80%homology with Euphorbia, barley, cucumber and tobacco, and shared greater than 70% homology with non 137
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京glutinous rice and soybean. The login ID in Genbank is EF617599. The result of our experiment and sequenceanalysis showed that the full length of the β-amylase gene cDNA is 1934bp, the opening reading frame is 459bp,which coded 152 amino acids, the Pi is 8.01 and the Mw is 16230.62. Our experiment could lay the foundationfor the further research on ramie β-amylase.S 3.01 - Transformation of cowpea (Vigna unguiculata L. Walp.) by Agrobacterium infiltrationAdesoye A1,2*, Togun A1, Machuka J31 Department of Crop Protection and Environmental Biology, University of Ibadan, Nigeria;2 Biotechnology Research Unit, International Institute of Tropical Agriculture (IITA), Ibadan, Nigeria.*Email: aadesoye@yahoo.comUntil recently, stable genetic transformation of cowpea through tissue culture technique could not be established.The aim of this work was to optimize inoculation and cocultivation medium factors for cowpea transformationand avoid in vitro regeneration procedures in obtaining transgenic cowpea through vacuum infiltration ofembryos.Using plasmid pCAMBIA 1301, influence of inoculation and cocultivation media compositions on transientgene expression were determined. Embryos were inoculated on Murashige and Skoog (MS) or yeast extractbroth (YEB) solutions supplemented with either acetosyringone, 0.05% silwet L-77, both or none and thencocultivated on MS solid medium for 24 hours. Gus assays showed that inoculating explants in MS containingacetosyringone and 0.05% silwet L-77 gave the highest transformation frequency (55.3%). When tested, thepresence of acetosyringone in the cocultivation medium increased transformation frequency by 15.35%. Whenuntransformed control seeds were screened on hygromycin and phosphinotricin 100% shoot and root growthinhibition were obtained at 50mg/l and 5mg/l, respectively. Secondly, cowpea embryos were transformed withAgrobacterium cells carrying two plasmids, ptjk 142 and pCAMBIA 1301, by inoculating them in mediaoptimized as above and subjecting to two rounds of vacuum infiltration. Thereafter they were cocultivated fornine days on MS selection-free medium where they germinated and produced mature plantlets. T1 seeds wereselected on antibiotic concentrations determined as above. Plants surviving both phosphinotricin andhygromycin showed gus positive reactions when subjected to gus histochemical assay and amplified the gusprimer during molecular analysis. Phosphinotricin resistant plants also amplified bar primers. Percentagetransformation based on total number of T1 seeds and number of plants with positive PCR reaction using bothprimers was 2.5 % for pCAMBIA 1301 and 3.9 % for ptjk 142 plants.The integration of transgenes into cowpea by Agrobacterium infiltration of embryos germinating on selectionfree medium has been demonstrated. This technique has the potential of rapidly producing stably transformedcowpea and avoiding all the limitations imposed by de novo regeneration in tissue culture.S 3.02 - Genetic Erosion in 2/12 Regions of Ethiopian Durum WheatAlganesh Tesema G.1,2, Giovanni G. Venderamin 4, E.Porceddu 2,31 Institute of Biodiversity Conservation and Research Addis Ababa, Ethiopia;2 International Doctoral Programme in Agrobiodiversity, Scuola Superiore, Sant Anna, Pisa, Italy;3 Dipartimento di Agrobiologia e Agrochimica, Universita degli studi della, Tuscia, Viterbo,Italy;4 Istituto di Miglioramento Genetico Piante Forestali, C.N.R. Via Atto Vannucci 13, 50134 Firenze, Italy. 138
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京*Email:Adishihu@yahoo.comEthiopia is an important centre of genetic diversity for Triticum durum. Genetic diversity is an indispensableresource for improving and stabilising crop yield. The objective of this study was to estimate the amount ofgenetic diversity loss. A total of 200 plants of durum wheat belonging to eight landrace populations, wereanalysed by two microsatellite markers per chromosome. Samples were collected at 15 years interval in tworegions of Ethiopia, Tigray and Shewa, the first severely affected by drought at different times, the other locatedclose to a business area. The average number of alleles per locus ranged from 2 to 10 with a mean of alleles perlocus of 4,35 in Koraro, Tigray, 1965; 15 years later the number ranged from 7 to 1 with a mean of 3.64,indicating that about one allele per locus got lost in that period. The number of alleles per locus was higher inShewa samples and an apparent lack of erosion was noticed. The number of alleles per locus was higher ingenome B than in A. The heterozygosity was higher at the beginning of the period analysed in Tigray (10% vs5%) than in Shewa (7% vs 5%) material; however the difference was statistically significant only in the Tigraymaterial but not in the Shewa one. In Tigray the first component accounted for 25.98% and the second for20.93% of the variation, whereas in Shewa the values were 27.09% and 21.63% respectively. In conclusion, thepresence of allele erosion in both regions, and erosion was more severe in Tigray than in Shewa.S 3.03 - Transformation of Brassica napus L. with an ihpRNA vector targeting the ∆-12 desaturase gene toachieve high oleic acid germplasmChen S*, Pu HM, Zhang JF, Gao JQ, Chen F, Long WH, Hu ML, Qi CKInstitute of Industrial Crops, Jiangsu Academy of Agricultural Sciences, Nanjing 210014.* Email chensong1963@126.comThe ∆-12 desaturase gene (fad2) is a key gene which contributes the most of the triacylglycerol polyunsaturatedfatty acid, such as linoleic and linolenic, in the seed of Brassica napus L. Down regulation of the activity of thegene in transgenic rapeseeds may results in an increase of oleic acid and decrease of linoleic in the seed. Anumber of methods have been reported to effectively silence the desaturase gene by gene transformation,including expressing fad2’s antisense, co-suppression, dsRNA or ihpRNA of fad2. Here we reported someprogress on the research of transgenic Brassica napus L.transformed with Agrobacterium harboring an ihpRNAvector targeting the fad2 gene.The transgenic oilseed rape (Brassica napus L) plants were obtained by Agrobacterium-mediated transformation.The cotyledon with approximately 2mm petiole from 4-day-old seedlings of Brassica napus L. cv. westar wereselected as explant for transformation. The cotyledon petioles were co-cultivated with Agrobacteriumtumefaciens strain LBA4404 harboring the vector pCNFIRnos, an ihpRNA vector. In its T-DNA region there aretwo expression cassettes: one is an inverted repeat unit of the 530bp segment of the fad2 gene with a spliceableintron sequence in between, controlled by a seed-specific napin promoter, another is a 35S-NPTII-Nos cassette,conferring kanamycin resistance. After being infected, the explants were sub-cultured on a shoot initiationmedium containing MS medium with 20mg/L kanamycin. After 4 to 6 week selective culture, kanamycinresistance green adventitious buds regenerated and were further rooted on the rooting medium. The resultingplants were transferred to soil to set seed.In 2008, the seeds of the T0 plant were harvested and the fatty acid composition of the seed was analyzed. The 139
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京result showed that some of transgenic lines have increased oleic acid content in seed oil compared to the control.Especially the line W-4 were found 7.65% increased oleic acid content compare to the non-transgenic controls.A year later, the selfed seeds of 49 T1 individuals from the transgenic line W–4 were harvested again and theseed fatty acids were analyzed by gas chromatogram further. The results showed that the variation range of oleicacid level among the T2 seeds of 49 progenies was 55.87% to 84.73% much wider than that of the control,and that of the control was about 55.28% to 67.43%. There were 19 individuals that contained greater than 75% oleic acid in the seed oil. including 11 individuals with more than 80% oleic acid. which accounted for 22.4% of the total plants. These high oleic acid lines were also found considerable reductions in linoleic acid andlinolenic acid. The oleic acid desaturation parameter(ODP)of the high oleic acid lines was only 5%-12% muchlower than that of the control. which had an ODP value of around 20%-36%. Based on the ODP value, it couldbe assumed that about 60%-83% fad2 activity in the seeds of high oleic acid lines was effectively inhibited,probably owing to the expression of ihpRNA for fad2 gene in the seeds of the transgenic plants, therebypreventing the oleic acid to be desaturated to linoleic acid and linolenic acid further. A southern blot analysis ofthe W-4 lines, probed with NPTII gene, showed that the W-4 lines contain a single copy of the transgene.S 3.04 - Agrobacterium-mediated maize C4-PPDK gene transformation of northern Japonica rice(Oryzasative L.)Cui ZH, Jia YG, Zhang LJ*, Fan JJ, Ruan YY, Zhu YS, Wang C, Hu KSchool of Biotechnology, Shenyang Agricultural University, Shenyang 110161.* Email: lijunzhang8@yahoo.com.cnRice (Oryza sative L) is one of the important food crops in China. The production of rice has a decisiveinfluence on food security. It is a dream for rice breeding expert to struggle for that select high-yielding varietyrice base on the super rice. Its not easy to get further increasing yield of super rice by traditional breedingtechnique. The hope will be true to increase the yield of super rice using high photosynthetic efficiency breedingtechnique by genetics engineering. So far the C3 pathway of rice could be changed into the pathway that hassome C4 pathway characters by transformation some photosynthetic key enzymes of C4 pathway, increasing thephotosynthetic efficiency and the resistance. In this paper, two varieties of northern Japonica rice of China,shennong265 and liaoxing1 were triumphantly induced the maize C4-PPDK gene by Agrobacterium-mediatedsystem. The system of transformation rice was set up under the condition of our current laboratory. Weidentified the resistance plants of the two rice varieties by Southern-blotting, and then we got some transgenicC4-PPDK rice plants. The effects of photosynthetic characters of transgenic rice were investigated. All in all, theincreasing of photosynthesis of the transgenic rice plant was not significance, but the expression of PPDK geneand the activities of PPDK increased available.S 3.05 - Effects of various modified strains of Agrobacterium tumefaciens on Genetic Transformation ofBanana and Plantain (Musa spp.L.)Esuola CO*1, Tripathi L2and Fawole I31 Fruits and Biotechnology Programme, National Horticultural Research Institute, P.M.B. 5432, Idi-Ishin Jericho,Ibadan, Nigeria;2 International Institute of Tropical Agriculture- Uganda; 3 Bells University, Otta, Ogun-State.* Email: bolajiius@yahoo.com 140
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Investigations were conducted to select the appropriate Agrobacterium tumefaciens strains for efficienttransformation of the banana cultivar ‘Williams’. Three A.tumefaciens strains (EHA105, C58, and GV2260)were used for the study. The ability of these strains to transfer β-glucuronidase gene into banana tissues wasassessed through GUS assay. Results obtained demonstrated that GUS activity was more enhanced in bananameristem pieces that were co-cultivated with EHA105 strain than in tissues that were co-cultivated with theother two strains. The least transient GUS expression was recorded in banana tissues that were co-cultivatedwith C58. The effect of the strains on efficient regeneration also indicated that EHA105 is more efficient thanthe other two strains. Therefore, EHA105 appears to be the appropriate strain for efficient transformation ofbanana.S 3.06 - The expression of ipt transcriptionally fused with AtGolS2 in frame of AtGolS2-ipt in transgenictobacco,plants showed drought-, salt-, cold-tolerances, and vigorous growthGuo JC 1 *, Hu XW 2, Duan RJ1 and Fu SP11 State Key Laboratory of Tropical Crops Biotechnology, Institute of Tropical Bioscience and Biotechnology,Chinese Academy of Tropical Agricultural Sciences, Haikou 571101, China2 Hainan University, 571104 Haikou ChinaCytokinins are a group of phytohormones with activities in many aspects of plant growth and developmentincluding cell division, initiation of shoot growth. ipt gene isolated from the plant pathogenic bacteriumAgrobacterium tumefaciens encodes an enzyme isopentenyl transferase, which catalyzes the rate-limiting stepof cytokinin biosynthetic pathway (Akiyoshi et al. 1984; Barry et al. 1984). Transgenic plants or tissues with iptunder control of constitutive promoter have been clearly associated with a marked increase in cytokinin content,which resulted in drastic changes in the morphological characteristics including non-elongated shoots, severelack of apical dominance leading to bushy phenotype, small round or serrated leaves, undeveloped roots,delayed leaf senescence, and sterility (Smigocki 1991; Mckonzie et al. 1994; Geng et al. 2001; van der Graaff etal. 2001).Gene fusion generally distribute to transcriptional and translational types. In transcriptional fusion, most ofprotein-coding sequences are derived from the reporter, few from the controller, although the mRNA mayconsist of controller and reporter. The reinitiating efficiency of the second gene translation in transcriptionalfusion is reduced by the terminal codon of the first gene and the out-frame of the internal nucleotides (Kozak etal. 1978; Bevan 1984; Rogers et al. 1985; Peabody et al. 1986; van Duijn et al. 1988; Joanna et al. 1989). In thisresearch, we combined AtColS2 gene with ipt gene in transcription manner of AtGolS2-ipt under the control ofCaMV35S promoter. In 35S- AtGolS2-ipt transcriptional fusion, AtGloS2 gene is closer to 35S promoter thanipt gene, thus AtGolS2 gene is vigorously expressed, whereas ipt gene is weakly expressed as the effects of theterminal codon of AtGolS2 and the internal sequence between the two genes (Fig.1B). The transgenic plantswill be suggested to enhance the tolerance to stresses resulting from the over- expression of AtGolS2,meanwhile, plants will have some vigorous growth characteristics as the ipt gene weaker expression.In 35S- AtGolS2-ipt transcriptional fusion, AtGolS2 gene was over-expressed, while ipt gene was weakerlyexpressed. The 35S-AtGolS2-ipt transgenic tobacco had a lot of promising characteristics including enhancingdrought-, salt-, cold- tolerances, and vigorous growth. The vigorous growth showed that the 35S- AtGolS2-ipt 141
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京transgenic tobacco grew faster with more roots at early stage under tissue culture conditions; or plants had larger,greener leaves, higher chlorophyll content, more flowers, delayed flowering, and delayed leaf senescence thanthe 35S-AtGolS2 transgenic tobacco under stresses, especially, cold stress. Thus ipt gene combining withanother gene, in which ipt is located in the secondary position of the transcription fusion, is a promising way forimproving plant growth. The expression of AtGolS2 –ipt transcriptional fusion in plants is function in planttolerances, especially, in cold tolerance.S 3.07 - Characterization and comparison of three transgenic Artemisia annua varieties andnon-transgenic variety in environmental release trialJiang LX 1,2,Liu H 1, Wang JB 1,2, Zhu H 1,2, Wu X 1,2, Tan FR 1,2, Zhao K 1,2, Tao SR 1,2, Tang KX 4, TangXM1,2,3*1 Biotechnology Research Institute, Shanghai Academy of Agriculture Sciences, 2901 Beidi Road Shanghai, ThePeoples Republic of China, 201106; 2 Supervision,Inspection and Test Center for Environmental Safety of GMCrops of MOA (Shanghai), 2901 Beidi Road Shanghai, The Peoples Republic of China, 201106; 3 KeyLaboratory of Agricultural Genetics and Breeding, 2901 Beidi Road Shanghai, The Peoples Republic of China,201106; 4 Plant Biotechnology Research Center, School of Agriculture and Biology, Shanghai JiaotongUniversity, The People’s Republic of China, 200240.*E-mail: saas.xmtang@gmail.comIn this study, three transgenic Artemisia annua varieties with contents of increased artemisinin were evaluated inenvironmental release trial. Firstly, because there are risks concerning genetically modified plants, the threetransgenic varieties were compared with non-transgenic plants in regard to agronomic traits, viability andexpressed material. Secondly, stress tolerances (salt, drought and herbicide) were evaluated under experimentalconditions. The results indicated there was no significant difference or significant difference between threetransgenic and non-transgenic plants in agronomic traits (as in plant height, crown width, stem diameter, leafshape, 1000-seed weight). Also, the viability (germination rate, transplant survival) from both transgenic andcontrol plants were similar. Moreover, it also was determined that three transgenic and control plants had similarstress tolerances (salt tolerance, drought tolerance, herbicide tolerance) and the expression of target genes intransgenic varieties was stable. Although the results of the environmental release trial indicated that thenon-transgenic and three transgenic A. annua varieties had no significant differences or significant differences inagronomic traits, viability and stress tolerances, as expected, the artemisinin content was increased in transgenicvarieties.S 3.08 - Generation of Marker-Free Transgenic Rice Carrying the Broad-Spectrum Blast Resistance GenePi9 via Co-transformation of Two Binary VectorsLi J 1, Wang X 1, Zhou J 1, Wang YL 2, Mao XQ2, He HY 1, Liu ZL 1, Li YL 1, Chai RY 2, Sun GC 2, Wang GL 3and Qu S 1,*1 Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China.Institute of Plant Protection and Microbiology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021,China. 2 Department of Plant Pathology, Ohio State University, Columbus, OH43210, USA.*Email: squ111@163.com 142
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Rice blast, caused by the fungal pathogen Magnaporthe oryzae, is one of the most destructive rice diseasesworldwide and destroys enough rice to feed more than 60 million people annually. Development of transgenicrice lines carrying cloned rice blast resistance genes is an effective method for control of the blast disease. Pi9 isa broad-spectrum blast resistance gene cloned in rice and has been transformed into rice cultivars fordevelopment of blast-resistant transgenic rice. To produce marker-free Pi9 transgenic rice, we utilized theapproach of co-transformation of two Agrobacterium binary vectors to introduce the cloned Pi9 gene into ricecultivar Zhejing 22 via. A 13.5-kb Pi9 genomic sequence was cloned in the T-DNA region of the first binaryvector (pLJ42). The T-DNA of another binary vector (pRB01) contained the Hygromycin resistance gene (HPT)as plant transformation selection marker. To facilitate screening of marker-free transgenic progeny, greenfluorescent protein (GFP) gene that was next to HPT in the pRB01 T-DNA was used as a visual marker forsorting out HPT/GFP marker-carrying T1 plants. Genomic DNA of 91 Zhejing 22, T0 plants was PCR analyzedusing primers specific to Pi9, T-DNA LB and RB borders, HPT and GFP. A total of 18 T0 plants carried both thepLJ42 T-DNA (Pi9) and the pRB01 T-DNA (HPT/GFP). T1 plants of transgenic lines carrying the Pi9 T-DNAwere inoculated with the Philippine rice blast isolate PO6-6 and 12 blast isolates collected in Zhejiang Province,China; blast-resistant Zhejing 22 T1 plants were obtained. Through PCR analysis of pLJ42 T-DNA (Pi9) andpRB01 T-DNA (HPT/GFP) of T1 plants, marker-free transgenic progeny plants carrying the Pi9 blast resistancegene were obtained from 2 independently transformed Zhejing 22 transgenic lines.S 3.09 - Overexpression of a wheat MYB gene increased multiple resistances in transgenic tobaccoLiu HX, Zhou XY, Dong N, Liu X, Zhang ZY*The National Key Facility for Crop Gene Resources and Genetic Improvement, Institute of Crop Sciences,Chinese Academy of Agricultural Sciences, P R China, Beijing 100081. **Corresponding author: ZengyanZhang. Tel: +86-010-82108781. Fax: +86-010-82105819,*E-mail: Zhangzy@mail.caas.net.cnMYB transcription factors are involved in plant development and a variety of stresses. A full-length cDNAsequence of a wheat MYB gene namely TaPIMP1was isolated from wheat (Triticum aestivum L.). The transcriptlevel of TaPIMP1 gene was up-regulated by inoculation with a fungal pathogen Bipolaris sorokinian. Enhancedaccumulation of TaPIMP1 was observed in wheat seedlings in responses to drought and exogenous ABAtreatments. TaPIMP1 gene encodes a MYB protein TaPIMP1, which contains two MYB DNA binding domains(R2, R3), two putative nuclear localization sites and two putative transcription activation domains. The entiresequence of the protein TaPIMP1 shares a low identity (<43.69%) with those of other plant MYB proteins.TaPIMP1 is a new member of R2R3 type of MYB transcription factors of transcription families. Transientexpression of GFP fused with TaPIMP1 in onion epidermal cells proved that subcellular localizations ofTaPIMP1 were the nucleus, confirmed that the TaPIMP1 is a transcription factor. To investigate the biologicalfunction of this gene, the gene transformation vector pBI-35S:TaPIM1 was constructed, and transferred intotobacco (Nicotiana benthamiana) by Agrobacterium mediated transformation method. After screening throughPCR and RT-PCR analysis on the target gene, transgenic tobacco lines over-expressing TaPIMP1 were obtained.The T1 plants of these transgenic lines were evaluated for pathogen resistance, drought and salt tolerance.Compared to the wild-type plants, TaPIMP1 overexpressing plants displayed significantly-enhanced resistance 143
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京to Ralstonia Solanacearum, and exhibited enhanced resistance to drought and salt stresses. In these transgenictobacco lines, activities of phenylalanine ammonia-lyase (PAL) and SOD were increased obviously than those inwild-type tobacco plants. These results suggested that the wheat R2R3-MYB transcription factor play animportant role in modulating responses to biotic and abiotic stresses.S 3.10 - Integrated Approach for Reliable Sample Archiving and DNA Extraction of PlantsQiu J, McCullough A, and Oommen AGeneSeek, a Neogen Company, 4665 Innovation Drive, Suite 120, Lincoln, NE 68521, USA.Email: jqiu@neogen.comReliable sample archiving and low-cost DNA extraction are critical for effectively carrying out practicalmarker-assisted plant breeding programs. We have developed a simple approach to archiving leaf samples usinga collection device and a simple, high-throughput DNA extraction procedure. Leaves are collected and placedin a small (5 x 5 cm2) collection device with printed barcodes. Two to three small (~2 mm diameter) dry leafpunches per sample were taken directly from the collection device, and the DNA extracted utilizing a modifiedalkaline lysis method. The DNA samples are suitable for routine PCR applications and SNP analyses usingMassARRAY® genotyping system. Our studies demonstrate that the archived leaf samples can be stored atambient temperature and DNA extracted from a wide variety of plant species give robust and reproducibleresults. This simple approach does not require expensive reagents or equipment and is applicable forhigh-throughput genotyping in a variety of plant species.S 3.11 - Studies on non-tissue culture plant genetic transformationSun Y, Wang JX, Yang LY, Cui GM, Du JZ, Hao YS and Wang YXAgri-Biotechnology Research Center of Shanxi Province, 030031 ChinaEmail: sunyi692003@yahoo.com.cnGenetic transformation is a powerful tool for plant breeding and genetical, physiological or biochemicalresearch. Two methods are currently widely used for producing transgenic plants, namely Agrobacteriummediation and particle bombardment. However, both of them need tissue culture procedures, which are labor-and time-consuming, tedious and expensive. We have been attempting non-tissue culture or in planta planttransformation methods for more than one decade and successfully obtained transgenic plants with followingapproaches.1. Pollen-mediated transformation of maize with assistance of ultrasonication the genetic transformation wasachieved by pollen-mediated approach on maize (Zea mays L.) inbred lines, Tai 9101 and Zong 31. PlasmidDNA of pCL II-RC-1 was mixed with fresh pollen of the maize inbreds in sucrose solution. The pollensuspension were treated with ultrasonication, collected, and pollinated on silks of maize ears. Transformantswere confirmed by dot bolt hybridization, PCR amplification and PCR-Southern blot hybridization (ActaBotanica Sinica 43: 275-279).2. Agrobacterium transformation of maize germinating seeds Germinating seeds of maize (Zea mays) werewounded in the meristem area using a scalpel, and co-cultivated with an Agrobacterium tumefaciens strainharbouring a Ti plasmid. Seedlings produced from the treatment were screened by hygromycin selection. Fertile 144
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京transgenic T0 and T1 plants were obtained. PCR amplification, PCR–Southern and Southern-blot analysisshowed that the foreign gene had been introduced into the inbreds of maize. About 29% of T0 seedlingsexamined were confirmed to be transgenic, although the overall transformation was only 0.6% when totaltreated seeds were taken into account (Biotechnology and Applied Biochemistry 46: 51–55).3. In situ transformation of poplar transgenic poplar shoots was obtained using Agrobacterium-mediated in situbud transformation. The method is demonstrated for the first time for poplar. Buds of Populus cathayana Rehdwere co-cultivated with an Agrobacterium strain LBA4404 harbouring the binary vector pBI101-Bmk-chi. Withthe procedure described, transformation efficiencies of 1% and 2.24% were achieved in 2006 and 2007,respectively. Stable integration of the transgene sequence was confirmed by PCR and Southern hybridization.The transgenes could be inherited by vegetative propagation. The novel approach is especially useful for speciesor cultivars not responding to tissue culture. (Scandinavian Journal of Forest Research 25: 3-9).Advantages of in planta transformation are: obviate tedious tissue culture procedures; rapid, thus time-saving;economical; labor-efficient; genotype independent; simple and ready to be integrated into conventional breedingprograms; and easily to be adopted in marker free transformation. Transformation efficiencies of the methodsvary between 1-40%. Studies for further enhancing transformation efficiency are under way.S 3.12 Factors Influencing on the Transformation Efficiency of Anthurium andraeanumWang F1, Liang FL1, Li YH2,Yi MS2, Xia Q2 and Zhang ZS11 Guangdong Provincial Key Laboratory of Plant Molecular Breeding, South China Agricultural University,Guangzhou 510642; 2 Guangzhou Flower Research Center,Guangzhou 510360, China.Email: zszhang@scau.edu.cnAnthurium andraeanum is a tropical flower with a unique flower pattern, elegant leaves and a long-lastingornamental period, with high ornamental and economic value. This flower can not only be enjoyed for pot plant,but also be used as high-grade cut flower, therefore, has a wide market prospect. Creation of new cultivars hasgreat importance of enhancing the development of flower industry, and transgenic breeding is a promising andan effective method to develop new varieties. Several studies about transgenic of Anthurium andraeanum havebeen reported, but the transgenic efficiency was generally lower. So, further studies on the factors influencingon the transformation efficiency of Anthurium andraeanum and establishing a high-efficient genetictransformation system have important significance for developing new varieties of Anthurium andraeanum bytransgenic breeding method.In this paper, calli were employed to investigate the factors influencing on the transformation efficiency ofAnthurium andraeanum by Agrobacterium-mediated method. results showed that pre-culture day, co-culture Thetime, the concentration of infection bacteria fluid and infection time had significant effects on the transformationefficiency of Anthurium andraeanum. In a certain range, increasing the pre-culture day, co-culture time,concentration of infection bacteria fluid and infection time improved the frequency of transient expression ofGUS gene. Acetosyingone was necessary for the genetic transformation of Anthurium andraeanum, thetransformation efficiency was significantly improved by adding acetosyingone in culture of Agrobacterium andthe medium of co-culture. The components of infection bacteria fluid also had significant effect on thetransformation efficiency of Anthurium andraeanum, the frequency of transient expression of GUS gene washighest when the Agrobacterium was suspended in the MS fluid medium. Different trauma modes had different 145
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京transformation efficiencies, the cutting resulted in higher transformation efficiency than acupuncture. Theoptimal conditions for transformation were that pre-culture time was 2 d, the suitable concentration ofacetosyingone in the bacterium suspension and co-culture medium both were 100 µmol·L-1, the OD600 ofinfection bacteria fluid was 0.4, the optimal time of infection and co-culture was 20 min and 3 d, respectively.The highest transient expression rate of GUS gene was 17.5%. Two plantlets were regenerated from 257 lumpsof hygromycin-resistant calli. The results of PCR analysis proved that the GUS gene was integrated to theAnthurium andraeanum genome.In the study,the genetic transformation system of Anthurium andraeanum was successfully established byAgrobacterium-mediated method. This lays a solid foundation on utilizing genetic engineering technology toimprove the traits of Anthurium andraeanum.S3.13 - Event-specific qualitative and quantitative PCR detection of the transgenic Bt rice line ZG22Wang XF, Chen XY, Miao QM, Fang J and Xu JFResearch Institute of Agriculture Quality and Standard, Zhejiang Academy of Agricultural Sciences, ShiqiaoRoad 198#, Hangzhou 310021, Zhejiang, People’s Republic of China.E-mail: njjfxu@163.comAn event-specific detection method was developed for the transgenic Bt rice line ZG22 which is not approved inany country so far but it is likely to be approved in China in the near future. The 3’integration junction betweenhost plant DNA and integrate DNA of transgenic ZG22 was isolated using thermal asymmetric interlaced(Tail)-PCR, and the specific PCR primers and TaqMan probe were designed based upon the revealed3’-integration junction sequence; in conventional qualitative PCR assay, the limit of detection (LOD) wasestimated to be 10 initial haploid copies. In the quantitative TaqMan real-time PCR assay, the limit of LOD andthe limit of quantification (LOQ) were approximately 1 and 5 initial template copies, respectively. There mixedrice samples with known ZG22 contents were used to verify the developed real-time PCR system, from whichthe expected results were observed.S 3.14 - Enhanced phosphorus uptake and Al tolerance in transgenic canola that overexpressed citratesynthase geneWang Y1, 2, Xu FS 1, 2*1 National Key Laboratory of Crop Genetic Improvement, Huazhong Agric. Univ., Wuhan 430070, China;2 Key Laboratory of Subtropic Agric. Resource and Environment, MOA, Huazhong Agric. Univ., Wuhan 430070,China.*Email: fangsenxu@mail.hzau.edu.cnLow phosphorus (P) availability and Aluminum (Al) toxicity are two major constraints to plant growth on manyagricultural soils. An increase in the excretion of organic acids, particularly citrate, has been identified as apotential mechanism to enhance P uptake and Al-tolerance in many plants species. Previous work reported thatexpression of a citrate synthase (CS) gene in plants resulted in the improvement of Al tolerance or Pacquirement from soils. In this study, we overexpressed a Pseudomonas aeruginosa citrate synthase gene incanola (Brassica napus cv Westar) and tested the P uptake and Al tolerance of transgenic plants. In Pi-limited 146
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京soil, the transgenic line CS3 accumulated much more biomass than wild-type plants (WT). The transgenic linesCS3 and CS6 also exhibited significant improvement in P efficiency with 32.9% and 26.6% increases in plant Pcontent, respectively, as compared with WT grown in Al-phosphate media. Furthermore, the transgenic plantsexhibited higher seed yields than WT when Al-phosphate was the P source in soil. The CS activity of transgenicplants was about two fold higher than WT with Al treatment or not. Citrate and malate are two importantorganic acids in tricarboxylic acid (TCA) cycle, and are also the main exudated anions in Al-exposed B. napus.Our results showed that the transgenic lines CS6 and CS3 secreted much more citrate and malate from intactroots than WT when exposed to 50 µM and 100 µM AlCl3, separately. The results suggested thatoverexpression of CS could be a useful tool to improve P uptake and Al tolerance in Brassica napus.S 3.15 - Variation of agronomic traits of transformed wheat by pollen tube pathwayXue XF, You J, Cheng TT, Zhang J, Liu JY, Ma J, Jia YY, Gao P, Wang S G, Wei QC, Zhang SL, Ru ZG, ZhouY*Department of Biotechnology, School of Life Science and Technology, Henan Institute of Science andTechnology, Xinxiang, 453003, Henan, China*Email: yanzhou@hist.edu.cnWheat transformation by pollen tube pathway has been widely applied in wheat genetic breeding. Accumulatingevidences have shown that variation of agronomic traits of transformed wheat by this technique could exhibit indifferent patterns and greatly relate to genotype and transformation protocol. In this study, five strains of wheat(BN-64, YM-34, KDZ-30, BM-1 and AK-58) were transformed with a plasmid construct carrying SeNHX1 geneby pollen tube pathway. The transformed plants were maintained in field under the normal condition and theiragronomic traits were examined when those plants grew to mature. The average of plant height in wheat (cm),ear length (cm), flag leaf area (cm2) and thousand- grain weight (g) measured in D1 generation of thetransformed plants were described as following: 64.77, 6.93, 31.63 and 31.5 in BN-64; 60.58, 7.54, 18.53,34.1 in YM-34; 84.77, 8.7, 30.15, 43.3 in KDZ-30; 48.17, 7.06, 29.30 and 33.4 in AK-58; 61.45, 8.37, 27.23,35.5 in BM-1, respectively. In D1 generation of those plants, compared with control, an obvious increase inthousand-grain weight has been found, and the variation of flag leaf areas showed the similar pattern, exceptBM-1; a decrease in plant height presented in YM-34 and AK-58 while an increase exhibited in the remainingstrains; ear length in BN-64 and YM-34 decreased and increased in KDZ-30, BM-1 and AK-58. Statisticalanalysis revealed that there was a significant difference found in plant length in D1 generation of transformedplants except KDZ, and a significant difference in ear length was observed in all the transformed plants exceptBM-1. There was no significant difference found in flag leaf area among all of the strains. Based on the resultspresented above, a suggesting conclusion can be made: an introduction of foreign DNA to wheat by pollen tubepathway could result in a variation of agronomic traits of the transformed wheat in different ways. Furthermolecular analysis is required to identify the transgenic wheat strains. This study may provide usefulexperimental data and technical support for wheat genetic breeding research.S 3.16 - The screening of general medium for indica riceYu Y, Zhang YS, Dong Y, Wan WT, Li LJ, Zhang J, Ling Q, Hu FYFood Crop Science Institute, Yunan Academy of Agricultural Sciences, Kuming, China. 147
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Indica rice don’t have a general induction medium.In order to find a general medium for indica rice,we designan orthogonal text which have three factor .The three factor are basic medium ,2.4 - D and 6 - BA .The basicmedium are MB, NB, N6, CC .Every hormones have 4 level. There are 16 combination been design.Everycombination have fixed nutrients (300mg/L hydrolyzed casein +500mg/L proline+250mg/L glutamine+30gsugarose+8g/L Agar)IR64,Minghui63,RD23 three varieties have been induce callus on these medium. Callusinduction rate and quality of every combination have been evaluated. From the result we get the three factors ofinfluence on callus induction and optimum combination.The 2.4 - D has the most important effect on callusinduction rate .The 6 - BA significant influence on the quality of callus.The combination of MB + 2.4 - D 5.0mg/L + 6 - BA 0mg/L + 300mg/L hydrolyzed casein+500mg/L proline +250mg/Lglutamine +30g/L sugarose+8g/LAgar has the best induction result for indica rice. The verification test show the medium has goodinduction results for most indica rice.S 3.17 - Application of Ac-Ds transposable element and fluorescent proteins in marker-free transgenicrice breedingZhou J1, Li J1, Wang X1, He HY1, Liu ZL1, Li YL1, Tu JM2, Wang YL3, Sun GC3, Wang GL4 and Qu S1,*1 Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021,China;2 College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310029, China;3 Institute of Plant Protection and Microbiology, Zhejiang Academy of Agricultural Sciences, Hangzhou310021, China;4 Department of Plant Pathology, Ohio State University, Columbus, OH43210, USA.*Email: squ111@163.comTo develop marker-free transgenic rice with improved insect/disease resistance, we have developed effectiverice transformation vector systems using the maize Ac-Ds transposable element and the green fluorescentprotein (GFP) gene. Initially, we constructed a T-DNA vector (pLJ26) containing a Ds element, Ac transposase,hygromycin resistance gene (HPT) and GFP. The Ds element functioned as a carrier of the BT insect-resistantgene or an RNA interference expression cassette conferring resistance to rice stripe virus (RSV). GFP was usedto track the HPT antibiotic selectable marker together with the Ac transposase and other T-DNA components.Rice varieties (lines) Zhejing 22, Zhehui H414, Kongyu 131 and Yuetai B were transformed with pLJ26 whoseDs element carried BT or the RNAi cassette as above. Molecular analysis of transgenic plants showed that 60%of the T0 plants underwent Ds transposition. In T1 generation, GFP was used as a visual marker to sort outmarker-carrying transgenic progeny and PCR assays of the GFP-negative T1 plants were performed to screenBT- or RNAi-containing Ds plants. Marker-free transgenic progeny were obtained from three BT-transformedrice lines and one RSV-RNAi-cassette transformed line. Using the Ac-Ds transposable element vectors, only asmall number of independently transformed rice plants were needed for the purpose of marker-free transgenicrice breeding because in a few rice transformants Ds element carrying the insect/disease resistance gene can bedistributed to multiple sites of the rice genome. Therefore, such a rice transformation event is equivalent to anumber of rice transformation events of the conventional co-transformation system. The Ac-Ds vector is more 148
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京suitable for marker-free transgenic breeding of the indica rice cultivars because indica rice transformation is lessefficient compared to japonica rice. In addition, we have utilized an improved co-transformation system toproduce marker transgenic rice of the cloned rice blast disease gene Pi9. A 13.5-kb Pi9 genomic sequence wascloned in the T-DNA region of one binary vector (pLJ42). The T-DNA of another binary vector (pRB01)contained the hygromycin resistance gene (HPT) as plant transformation selection marker. To facilitatescreening of marker-free transgenic progeny, GFP next to HPT in the pRB01 vector was used as a visual markerfor sorting out HPT/GFP marker-carrying T1 plants. Genomic DNA of T0 plants was PCR analyzed usingprimers specific to Pi9, T-DNA LB and RB borders, HPT and GFP to screen individuals carrying both the pLJ42T-DNA (Pi9) and the pRB01 T-DNA (HPT/GFP). In the T1 generation, through PCR analysis of pLJ42 T-DNAand pRB01 T-DNA of T1 plants, marker-free transgenic progeny plants carrying the Pi9 blast resistance genewere obtained from two independently transformed Zhejing 22 transgenic lines. These results shows that Ac-Dstransposable element and GFP in transgenic rice can help improve the efficiency of genetic and molecularanalysis of transgenic plants, thus expediting the process of marker-free transgenic rice breeding.S 4.01 - Molecular Diversity on Ethiopian Tetraploid wheatAlganesh Tesema G1,2, Venderamin GG4, Porceddu E2,31. Institute of Biodiversity Conservation and Research Addis Ababa, Ethiopia;2.International Doctoral Programme in Agrobiodiversity, Scuola Superiore, Sant Anna, Pisa, Italy;3.Dipartimento di Agrobiologia e Agrochimica, Universita degli studi della,Tuscia, Viterbo,Italy;4.Istituto di Miglioramento Genetico Piante Forestali,C.N.R. Via Atto Vannucci 13, 50134, Firenze, Italy.Email: Adishihu@yahoo.comEthiopia has the second largest wheat growing area in Sub Saharan Africa, having 877,000 ha of arable landoccupied by wheat. A total of 30 populations of 790 individuals of Triticum durum originated from ten regionswere assayed using a set of 14 wheat microsatellites, representing 7 markers from genome A and 7 from genomeB located on 14 different chromosome locations. A total of 347 alleles were detected with average alleles of 24.8per locus. The highest number of alleles per locus was detected in the A genome with 34, 32, 31, 27, 25, 21, 20alleles per locus, compared to 30, 29, 16, 12 alleles per locus for genome B, respectively. The highest geneticvariation exists in the non centromeric regions than in the centromeric regions of chromosomes, in both GenomeA and B short arms. The mean of polymorphism of all over region were ranged, (100% - 92.86%).The analysisof molecular variance, revealed that 72% of the total variation was found within population, 12% amongpopulation and 16% among regions. High levels of genetic diversity were existed among regions (17%) ingenome A long arm. The highest variation of among populations were detected in Shewa, Bale and wello (21%,15 %,( 14%). whereas in Hararghe, Gojam & Tigray among populations were low (9%). But, the withinpopulations were existed in high level of diversity in (Tigray, Gojam, Hararghe), Gonder and Arsi, ((91%), 90%,& 88) %. The highest grand mean of observed heterozygosity were detected in Tigray, Hararghe, Shewa andWellega (8%, 7%, 6%, 6%).The highest grand mean diversity were found in Gojam, Wello, Hararghe,Arsi andwellega (4.1,4,3.9, 3.8, 3.8). while in shewa ,was obtained lowest (2.8), In general, In one or in another way ofparameters, every regions were exhibited significantly higher polymorphism. 149
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京S 4.02 - Genetic Diversity Analysis of Chinese Wild Musaceae based on SSR MarkersFeng HM 1,2 , Chen Y 1 , Li B 2,3, Deng CJ 2,3 and Wu YT 1, 2*1 Qiong Zhou University, Sanya 572022, Hainan Province, The People’s Republic of China;2 Key Laboratory of Tropical Crop Biotechnology, Ministry of Agriculture, Institute of Tropical Bioscience andBiotechnology, Chinese Academy of Tropic Agricultural Science, 4 Xueyuan Street, Longhua District, Haikou571101, Hainan Province, The People’s Republic of China;3 Hainan University, Haikou 570228, Hainan Province, China.Email: wuyaoting@tsinghua.org.cnThe Musacaea family including 3 genus of Musa、Ensete and Musella. Musa contains approximately 40 wildspecies which are distributed over the tropics area of Asia and Australia. Ensete contains 6-7 species which aredistributed tropics area of Asia and Africa. Musella was the unique genus in China. Musa was divided into threesections, Eumusa, Rhodochlamys and Callimusa by 12 wild species in China. Additional 1 Ensete species and 1Musella species were distribute in China. This paper using SSR markers annlysis the genetic diversity amongChinese Musaceae. SSR (Simple Sequence Repeat) analysis from 13 primer pairs. The number of polymorphicalleles generated ranged from 3 to 9 per primer; the PIC (Polymorphism Information Content) values were0.28-0.82, average 0.59. Similarity coefficients were calculated based on 63 selected polymorphic alleles andUPGMA clustering analysis was performed. The high levels of SSR polymorphism detected in this studyindicated the genetic relationships of wild Musa species in China. Additional SSR bands found in showed thatall 15 materials were divided into 3 groups on the level of similarity coefficient 0.54, Musella lasiocarpa ingroup I, Ensete glaucum in group Ⅱ, and other Musa species in group III. Group III can be divided into 4subgroups on the level of similarity coefficient 0.75. The first subgroup include 2 species and 2 subspecies fromsect. Rhodochlamys, M.Chunii, M. aurantiaca and M. sanguinea subspecies. The two subspecies of M.sanguinea were divided into 2 subgroups on the level of similarity coefficient 0.78. The second subgroupinclude 3 species from sect. Eumusa, M. acuminata var.chinensis, M. yunnanensis and M. sp. The new speciesM. sp. which was discovered by the second author in Yunan formed a distinct branch. The third subgroupinclude 4 species from sect. Eumusa, M. nagensium, M. itinerans, M. balbisiana and M. basjoo. The fourthsubgroup include 2 species from sect. Callimusa, M. coccinea and M. paracoccinea. On the level of similaritycoefficient 0.84, all the materials can be distinguished from each other.S 4.03 - Analysis of genetic diversity and SSR allelic variation in rubber tree (Hevea brasilensis)Feng SP1,4, Li WG2,3, Yu F1,2, Wang JY1,2, Feng HM4, Wu YT4*1 Key Laboratory of Tropical Crop Biotechnology, Ministry of Agriculture, Institute of Tropical Bioscience andBiotechnology, Chinese Academy of Tropic Agricultural Science, 4 Xueyuan Street, Longhua District, Haikou571101, Hainan Province, The People’s Republic of China.2 Key Laboratory of Tropical Horticulture Plant Resources & Genetic Improvement, Ministry of Education,Agricultural College, Hainan University, 58 Renmin Avenue, Haikou 570228, Hainan Province, The People’sRepublic of China.3 Key Laboratory of Rubber Biology of Ministry of Agriculture, Rubber Research Institute, Chinese Academy ofTropical Agricultural Sciences, Danzhou City 571737, Hainan Province, The People’s Republic of China.4 Qiongzhou University, Sanya 572002, Hainan Province, the People’s Republic of China 150
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京16 SSR (simple sequence repeat) primer pairs amplified across 45 cultivars, 11 wild accessions of rubber tree,and 3 related species. the results showed: a total of 43 alleles were obtained from 10 EST-SSR (expressedsequence tag derived simple sequence repeat) primer pairs across cultivated and wild accessions, with anaverage of 4.3, PD (power of discrimination) value ranged from 0.420 to 0.834, with an average of 0.597, a totalof 30 alleles were obtained from 6 genomic SSR (gSSR) primer pairs, with an average of 5, PD value rangedfrom 0.587 to 0.772, with an average of 0.689, HBE280, M197 and MnSOD were the most informative; 16 SSRprimer pairs amplified with an average of 4.38 alleles in cultivars, Ho = 0.587, and with an average of 4.13alleles, Ho=0.568 in wild accessions; in comparison with every two materials of 45, GS (genetic similarity)value ranged from 0.51 to 0.95 between cultivars, which ranged from 0.48 to 0.79 between cultivars and wildaccessions; UPGMA ( unweighted pair group method with arithmetic ) cluster analysis based on the GS valueshowed: all the cultivated and wild accessions were clustered into a big category, on the level of similaritycoefficient 0.68, wild accessions were distinguished from cultivars; 61 sequences were sequenced from 5EST-SSR loci, of which 44 sequences belonged to AG/TC repeats, in comparison with the original ESTsequences, insertion, deletion, conversion and transversion mutation occurred in SSR repeats and flankingregions, long repeat sequences had more variations, moreover, point mutation frequently occurred in flankingregions indicated the new SSR loci in rubber tree.S 4.04 - Studies on genetic relationship and diversity for Malus robusta Rehd. by TP-M13-SSR analysisGao Y, Wang K, Gong X, Liu LJ, Liu FZ *Research Institute of Pomology, CAAS/Key Laboratory of the Use of Fruit Germplasm Resources, Ministry ofAgriculture, Xingcheng 125100, ChinaEmail: apgr@vip.163.comSimple sequence repeat (SSR) detection with tailed primer M13 (TP-M13-SSR) was used in studies on geneticrelationship and diversity for Malus robusta Rehd. Resources collected.16 SSR markers amplified 205 alleles,with an average of 12.8 alleles per locus. Except M.baccata(L.)Borkh,M.prunifolia(Willd.)Borkhausen andM.domestica Borkh., the average of genetic diversity, locus heterozygosity and PIC for 31 cultivars of Malusrobusta Rehd.on 16 SSR locuses were 0.7879,0.8413 and 0.7592 respectively.The similarity coefficient betweenthe cultivars of Malus robusta Rehd. from Huailai of Hebei province and M.baccata(L.)Borkh,M.prunifolia(Willd.) Borkhausen was higher than those from Xiaofanshan, Dagucheng and so on. So the formerin genetic relationship may be closer to the latter two. Malus robusta Rehd. from Sankuaishi maybe later thanother types in the origin phylogeny.UPGMA cluster analysis showed that all the materials could be classifiedinto two groups,correlatively with origin and evolution.The first category consists mainly ofM.baccata(L.)Borkh and M.prunifolia(Willd.) Borkhausen,and the second category consists mainly of localvarieties of Malus robusta Rehd.. Huailai of Heibei province may be the mainly primitive origin, and those fromXiaofanshan area are more recently in genetic relationship than those from other areas.S 4.05 - Establishment of SSR fingerprints of Chinese new pear (Pyrus L.) cultivars and its application oncultivar identificationGao Yuan, Tian Lu-ming, Liu Feng-zhi, Cao Yu-fen * 151
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Research Institute of Pomology, CAAS/Key Laboratory of the Use of Fruit Germplasm Resources, Ministry ofAgriculture, Xingcheng 125100, China* Email : yfcaas@263.netThe fluorescent labeling of simple sequence repeat (SSR) detection was used to establish SSR fingerprints of 93new pear (Pyrus L.) cultivars.10 pairs of SSR primers screened amplified 132 alleles, with an average of 13.2alleles per locus. The range of observed heterozygosity was 0.2421~0.8632. The amplified banding patterns of10 loci were 8~44 with an average of 29.7. Each cultivar had its cultivar-specific SSR patterns and can use it as areliable method for cultivar identification. Two pairs of primers (KA16 and CH05d04) with high polymorphismcould distinguish all of the 93 pear new cultivars except for the mutants.S 4.06 - Genetic Analysis of Sweet Sorghum and Other Sorghums Using SSRsJi GS1*,Song YF1, Liu GQ1, Hao FW 21 Millet Crops Institute, Hebei Academy of Agri. & Forest. Science / National Millet Crops Improvement Centerof China , Shijiazhuang, China, 050031,2 University of Hebei Broadcast TV, Shijiazhuang, 050051* Email jigs0312@126.comSorghum (Sorghum bicolor L. Moench) is one of the most important crops in the world. Sweet sorghumpossesses higher ethanol production ability than corn and sugarcane for bearing high biomass and sugarenriched stalk. As a bio-fuel crop, sweet sorghum breeding has been an important project since “The EleventhFive-year Plan ” in China. Understanding the relationship between sweet sorghum and other kinds of sorghumsand finding sugar content related molecular markers are important for sweet sorghum breeding. The objective ofthis study by using SSRs is to determine the diversity of sweet sorghums, to identify the relationship betweensweet sorghums and grain sorghums, to screen the sugar content associated markers, and to offer moreinformation for sweet sorghum breeding.Thirty sorghum accessions including 19 of sweet sorghums, 10 grain sorghums and 1 wild sorghum, wereanalyzed using 63 pairs of simple sequence repeat (SSR) markers which distributed widely across the sorghumlinkage groups (A-L). The results showed that SSR markers were highly polymorphic among the sorghumcollections and the average alleles per primer were 2.76. The PIC ranged from 0.064 to 0.746 with the averageof 0.487. The accessions in this study were unequally separated and were clustered into six groups. Sweetsorghums were scattered into four groups and were significantly separated from the wild line (group2) and thelines from Hebei Agricultural University (group 1). The five R-lines and three B-lines were placed in group 3and 5 together with sweet sorghums. Group 4 and 6 were two sweet sorghum clusters and the lines from ChinaSci. Academy were clustered together. Six SSR markers were found related to sugar content (brix) by multipleregression analysis. These markers were different from the previous studies. The information of this studyshould benefit sweet sorghum breeding program and the sugar content related markers could be used for markerassistant selection.S 4.07 - SRAP Analysis of Genetic Diversity of Distylium chinense in Three Gorges Reservoir Region andEvaluation for Conservation Strategy 152
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Li XL, Li N, Yang J, Yang WH, Chen FJBiotechnology Research Center and Hubei Province Key Laboratory of Natural Products Research andDevelopment, China Three Gorges University,Yichang,Hubei 443002, China.* Email : lixiaolinggz@126.comDistylium chinense (Franch.) Diels, a member of the genus Distylium of the family Hamamelidaceae, is a shrubor small tree with high flooding tolerance. This species is a dominant riparian species and native to thewater-level-fluctuation zone in the Three Gorges reservoir of the Yangtse River, China. However, during thepast several decades, the natural habitat of Distylium chinense has suffered major destruction as the result ofanthropogenic activities such as construction of Three Gorges Dam (TGD) and floodgates on the rivers, excessreclamation for agriculture use, and excess firewood consumption by humans, etc. Therefore it is extremelyurgent to its protection, research and utilization.The present paper studied the genetic diversity and the geneticstructure of four in situ populations and one ex situ conservation population of D. chinense in the Three GorgesReservoir area using sequence-related amplified polymorphism (SRAP) molecular markers technique. A highgenetic diversity was observed in this species, with 7 SRAP primers generating a total of 46 scorable fragments,37 (80.43%) of which were polymorphic, the observed number of alleles (A) =2.00, the mean effective numberof alleles (Ae) =1.34, Nei’s gene Diversity index (Hp)= 0.2159 and Shannon’s Information index(I ) =0.3509,which were higher than the average value of seven endemic plant species. The total genetic diversity (Ht) forfive populations was 0.2188, the genetic diversity within population (Hs) was 0.1934, and the coefficient of genedifferentiation (Gst) was 0.1161, which indicated that 88.39% of the genetic variation occured within thepopulations and only 11.61% among the populations. The gene flow (Nm) was 3.8072, which implied that thegene flow was enough to maintain gene exchange and current genetic structure. Using un-weighted pairgroupmethod arithmatic average(UPGMA) and principal coordinates analysis(PCA), Letianxi population,Xiangxi population, Yanduhe population and Three gorges botanical garden population, of which growing alongriversides in the Three Gorges Reservior area, were clustered into one group; the Gaojiayan population far awayfrom riversides in the Three Gorges Reservior area was clustered into another group. The mean genetic diversityin ex situ conservation population was higher than that in situ populations, so the ex situ conservation programat Three Gorges Botanical Garden was considered to be successful in preserving genetic diversity. Combininganalysis of the genetic diversity with available information on breeding system, status of natural habitat andevolutionary history, we also evaluated the current ex situ conservation program and proposed some effectivestrategies to further conserve the species.S 4.08 - Assessment of Genetic Diversity and Relationship of Tea Germplasm in Yunnan as Revealed byISSR MarkersLiu BY, Wang YG, Li YY, Tang YC, Wang PSTea Research Institute of Yunnan Academy of Agricultural Science, Menghai 666201, China.E-mail:liusuntao@126.comTea is an important beverage crop in the world. Yunnan region in China is the origin center of tea plants. Ourknowledge on genetic diversity and relationship of tea germplasm in Yunnan province is critical to guide teabreeding. In order to provide theoretical information for using tea germplasm in tea breeding, we investigated 153
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京the relationship and genetic diversity of the tea germplasm in Yunnan province. Eight species, inculding 134 teavarieties were used to detect the genetic variation by inter-simple sequence repeats (ISSR) analysis. The resultshowed that 475 DNA fragments among all 134 tea accessions were amplified, using 18 reliable ISSR primers,among which 470 DNA bands were polymorphic (PPB=98.9%). This indicated that a great amount of geneticpolymorphism exists among tea germplasm tested. The genetic similarity (GS) among the tested genotypesranged from 0.445 to 0.819, with an average of 0.512, indicating a wide gene pool among tea varieties inYunnan. The cluster analysis presented that these resources were divided into three main groups using theunweighted pair-group method with arithmetic average (UPGMA) based on ISSR molecular marker data, butthe dendrogram did not indicate clear division among tested varieties based on their geographical origin.Principal Component analysis (PCA) for ISSR data showed that PCA supported UPGMA clustering result, butshowed more explicit relationships among the test accessions with different lays, orientations and positions. TheGS among 8 populations ranged from 0.850 to 0.987, with an average of 0.92, indicating that there existed asmall variation of genetic diversity among different population. The findings of this research would be favorablefor the further practice, such as tea breeding, the molecular genetic linkage mapping and the DNA fingerprintbuilding of tea germplasm.S 4.09 - 辽宁地区水稻区域试验品种遗传多样性分析马慧 1 2 张丽 1 陈丽静 1 钟鸣 1 郭志富 1 张立军 1 邵国军 2* ,1 沈阳农业大学辽宁省农业生物技术重点实验室 辽宁沈阳 110161, 2 辽宁省农科院 辽宁沈阳 110101 以辽宁地区的 2007 参加区域试验的 65 个水稻区域试验材料为研究对象,利用 SSR 分子标记技术对其遗传多样性和亲缘关系进行分析,从而分析材料间的遗传基础,为选育遗传基础广泛的水稻材料提供指导。从 200 对水稻 SSR 引物中挑选扩增条带清晰、带型稳定、差异显著的 18 对 SSR 引物进行进一步的遗传多样性比较分析,共检测出 83 个等位基因位点,平均 4.611 个,平均 Nei 基因多样性指数为0.529405,变幅为 0.219172-0.793506。结果表明:65 个水稻品种可划分为两大类群和三个亚类群,17 个小群,其中以第 4、5、6、7 类的品种有 43 个,占全部品种的 66%。S 4.10 - Analysis of Genetic Background of Dongzao (Zizyphus jujuba Mill. ‘Dongzao’) AdvancedSelections by AFLP MarkersMa QH, Wang GX*, Liang LSResearch Institute of Forestry, Chinese Academy of Forestry/Key Laboratory of Tree Breeding and Cultivation,State Forestry Administration, Beijing 100091, China.* Email: wanggx0114@126.com, mqhmary@sina.comDongzao (Zizyphus jujuba Mill. ‘Dongzao’) is one of the traditional excellent fresh-eating cultivars in Chinesejujube species which become mature in middle October. The original provenance of Dongzao derives from thebohai bay area in China where Hebei and Shandong provinces are jointed. Present researches and investigationsshowed that Dongzao trees planted in Hebei and Shandong provinces belonged to the same cultivar, butcontaining abundant and diverse variation in germplasm level. So the main work to effectively utilize theprecious germplasm resources should be concentrated on the analysis of genetic relationship, the separation ofdifferent traits, and the selection of advanced individuals in this cultivar. In this study, the collection of advanced 154
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京selections was based on the precise investigation in the orchards of Hebei and Shandong provinces in October,2006 and 2007. The phenotypic traits in the selection included the fruit shape, size, appearance, nutritionalquality and the yield. To analyze the genetic background of these advanced selections, the amplified fragmentlength polymorphism (AFLP) was performed. Compared with isoenzyme and RAPD polymorphism, AFLPcould provide more abundant polymorphic information. However, we had not found any reports on theidentification of Dongzao advanced selections with AFLP markers. In this study, genetic background of 52Dongzao advanced selections and their control, Jinsixiaozao and Jianzao cultivar from Hebei and Shandongprovinces were analyzed, using AFLP markers with 16 pair of Mse I/EcoR I primer combinations. Clusteringanalysis was performed by using of unweighted pair group method with arithmatic mean (UPGMA) methodbased on the simple matching coefficient (SM). The results showed that a total of 565 bands were amplified by16 primer pairs, of which, 225 were the polymorphic bands including Jinsixiaozao and Jianzao cultivar, and 99were the polymorphic bands among Dongzao advanced selections. There was some difference among the groupof Dongzao advanced selections (the average similarity coefficient was 0.9792), though the diference was lessin contrast with Jinsixiaozao or Jianzao cultivar (the similarity coefficient were 0.8294 and 0.7977, respectively).The three cultivars were divided into 3 groups and the group of Dongzao advanced selections were divided intoseveral parts in the cluster analyzing map. The genetic distance of the old Dongzao trees and the Dongzaoadvanced selections from Huanghua county, Hebei province was higher than the ones from Binzhou city,Shandong province, the similarity coefficient were 0.9649 and 0.9946, respectively. But there were someindividual candidates of aberrance in the latter. The results mentioned above showed that there was somevariation in Dongzao cultivar, and the eximious traits of the advanced selections were not all attributed to thedifferent growing condition and cultivating methods. So the results were much helpful for the geneticimprovement and the advanced selection of Dongzao cultivar.S 4.11 - Plantain (musa sapientum l) molecular diversity reveals narrow genetic base of local GhanaianaccessionsMarian D. Quain*, and Beloved DzomekuCouncil for Scientific and Industrial Research – Crops Research Institute, P.O. Box 3785, Kumasi, Ghana.* Email : marianquain@hotmail.comPlantains are important sources of high-calorie energy in Ghana. They are also of great socio-economicimportance in the country. Plantains are also very important sources of rural income. However, plantainproduction has been saddled by several constraints among which include pests and diseases and lack ofimproved technologies. As regards to genetic study, morphological characterization has identified about 100local accessions to exist in Ghana. However, no molecular characterization has been done on plantains.Characterization based on morphologic characteristics alone may be limited since the expression of quantitativetraits is subjective to strong environmental influence. Alternatively, molecular characterization techniques arecapable of identifying polymorphism represented by differences in DNA sequences. The objective of thisresearch was therefore to conduct molecular characterization of Ghanaian local accessions of plantain. Thisstudy project sampled 38 accessions of plantains in Ghana from the Ashanti and Eastern region. Samplingcovered all known morphologically categories. Molecular characterization was carried out on the assembledgermplasm. Simple Sequence Repeats (SSRs) were used to assess diversity. The SSR marker revealed that 24% 155
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京of the plantain accessions have a genetic distance of zero units. “Asamienu” was most distantly related to all theother varieties. An introduced hybrid Fhia 21 was also distantly related to all the local accessions, however, itwas found to be closely related to “Apantu” Red and “Apantu” Dichotomy. “Orishele Eumusa” an AABPlantain of Nigerian origin was related at distance Zero to some of our local varieties. Whereas “YadeEumusa” an AAB Plantain of Cameroon origin was closely related to some of the local accessions. Thedendogram generated two major clusters. “Asamienu” was distantly related to all assessions whereas FHIA 21closely related to two of the local “Apantu” collections. The SSR data pointed out that the plantain accessions inGhana are very closely related and hence breeding programs need to focus on broadening genetic base. This isthe first report on the use of molecular marker tools to carry out diversity studies on plantains in Ghana. Theinformation generated will be used as a document to encourage breeders to work towards broadening geneticdiversity in plantains.S 4.12 - Identification of high molecular weight glutenin subunits in Aegilpos crassa using SDS-PAGE andmass spectrometryNaghavi MR, Hassani ME, Aghaee MJ, Bamneshin M, Taleie ARAgronomy and Plant Breeding Dept., Agricultural College, University of Tehran, Karaj, Iran; HorticulturalDept., Agricultural College, University of Tehran, Karaj, Iran; Seed and Plant Improvement Institute, Karaj,Iran.* Email : mnaghavi@ ut.ac.irGlutenins are the most widely and intensively studied groups of wheat seed proteins. In this study 120 Iranianaccessions of Aegilops crassa collected from various geographical areas of Iran, were evaluated for variation inhigh molecular weight glutenin subunit compositions. In the most accessions, subunits showing electrophoreticmobility similar to that of Dy12 were present. Eleven allelic variants were observed in Glu-D1 locus with thehighest (30.90%) and the lowest allele (0.5%) frequencies in 3+12 and 2+10 variants, respectively. Among 17bands selected for MALDI-TOF-TOF-MS analyzing only 6 bands were identified with high probability and 11of them had no MS/MS data. The results showed that Iranian accessions of Aegilops crassa formed aninteresting source of favorable glutenin subunits that might be very desirable in breeding programs forimproving bread wheat quality.S 4.13 - Breeding high oil groundnut varietiesNigam SN, Venuprasad R and Udaykumar MIndia is a net importer of vegetable oil with more than 50% of the demand met through imports. Demand foredible oil, due to growth in population and economy, is projected to grow to 16.5 million in 2011 and 20.8million tonnes by 2015. Thus to meet the growing demand and also to reduce the burden on the stateexchequer it is important to boost the production of edible oil in India.Groundnut (Arachis hypogaea L.) is a major oilseed legume crop in India; it meets >30% of the requirement ofedible oil. The oil content in the currently cultivated groundnut varieties is about 48%. Even a slight increasein the oil content in groundnut seeds would significantly increase the profitability of the oil mills and thus theincome level of the farmers. Newer groundnut varieties with oil content higher than in the varieties currently 156
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京cultivated by farmers are needed to boost the edible oil production in India and correspondingly increase theincome levels of the farmers.There has been very little focused effort on breeding for high oil in peanut mainly due to lack of effective andeconomical breeding tools. As such any improvement in oil production has been brought indirectly outthrough expanding peanut production area and/or improving kernel yield per unit area. As both the aboveoptions have reached a point of saturation there is need to explore and develop ways to increase peanut oilproduction by improving oil synthesis per unit seed mass through new varietal and molecular approaches.Conventional breeding for high oil is slow, tedious and expensive, hence the use of molecular markers isexpected to increase efficiency and reduce costs. Such a tool is essential to sustain long term gains in oilproductivity. However, in groundnut, conventional genetic analysis to develop molecular markers is greatlyimpeded by paucity of polymorphic markers. The candidate gene based approach to develop markers for highoil trait is an innovative alternate approach. Potential candidate genes could be obtained from knownbiochemical pathways for oil production in plants and from publicly available expressed sequence tag (EST)databases. Since oil biosynthetic pathway is well characterized, it is possible to identify superior alleles of keygenes (Ohlrogge et al. 2000; Hobbs et al. 2004) that could be used in breeding. Details of this approach arediscussed.S 4.14 - Quantitative Genetics and Heterosis Study of Agronomic Traits in Autotetraploid RiceShahid Muhammad Qasim1 , Xu HM 2 , Li JQ 1 , Li YJ 1 , Wang SZ 1 and Liu XD 1*1 Guangdong provincial key laboratory of plant molecular breeding, South China Agricultural University,Guangzhou, Guangdong, 510642, China.2 Department of Agronomy, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310029,China.* Email:xdliu@scau.edu.cnGenetic effects and genotype by environment interaction for important agronomic traits of autotetraploid ricewere analyzed by additive, dominance and additive × additive model. It was shown that genetic effects hadmore influence on agronomic traits of autotetraploid rice than genotypic environment interaction. Plant height,panicle length, seed set, yield, dry matter production and 1000-grain weight were mainly controlled bydominance variance. Additive and additive × additive gene action constructed the main proportion of geneticvariance for heading date (flowering), number of panicles, grains per panicle, grain length, however grain widthwas supposed to be affected by additive × additive and dominance variance. Heading date produced highlynegative heterosis over mid parent (Hpm) and better parent (Hpb), whereas Hpm and Hpb were found to be highlypositive and significant for yield, seed set, peduncle length, filled grains and 1000-grain weight in F1 and F2generations. Autotetraploid hybrids 96025 × Jackson (indica/japonica), 96025 × Linglun (indica/indica) andLinglun × Jackson (indica/japonica) showed highly significant hybrid vigor with improved seed set percentageand yield. These results suggest that autotetraploid rice has the potential to be used for further studies andcommercial application. 157
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京S 4.15 - A genome-wide scanning for association marker loci with female fertility and spike grain numberin wheat---from small diverse extreme populationTong T1, 2 , Dou BD*1,2, Zhu XB1,2, Zhang XL21 Jiangsu Key Laboratory for Eco-Agricultural Biotechnology around Hongze Lake,Institute of plant biotechnology, Huaiyin Normal University, Huai’an 223300, Jiangsu, China;2 Agronomy College, Xinjiang Agricultural University, Urumqi 830052, Xinjiang, ChinaEmail : doubd@163.comIt is a tough task to scan gene loci in wheat genome associated with a trait. In order to find more gene markerloci efficiently associated with female fertility and spike grain number in wheat. A small populations with 60variety lines called ‘S involved (female sterile line S) diverse extreme population’ were composed as testedpopulation, 42 polymorphic SSR markers in wheat genome were selected as background markers to test its’population genetic structure, MLM model in software TASSEL was used to check 210 polymorphic markers’association degree with phenotype Female Fertility (domestic fertility---D.F and International fertility---I.F.) in2 years, the highly associated markers with phenotype from small diverse extreme population analysis wereverified in relatively large population of S involved 261 variety lines. Resemble method was used to selectassociation markers with phenotype spike grain number. The result showed that there was not obviouslyLinkage Disequilibrium between background markers. 3 sub-populations were found in the small populations.24 or 28 markers were found associated with both of the two types of female fertility values in 2 years by small‘S involved diverse extreme population’, 21 or 23 markers were verified association with both two types offemale fertility values in 2 years by large ‘S involved 261 variety lines population’. 8 markers among 21 or 23female fertility related markers were also associated with spike grain number in wheat. The phenomena thatfemale fertility association marker locus (such as Xwmc25, Xgwm71, Xgwm30…) had its’ effect on spike grainnumber were also existed on the 2DS in our previous linkage analysis.S 4.16 - Genetic Diversity and DNA Fingerprint Analysis in Wild, Weedy and Cultivated Azuki Bean(Vigna angularis) Using SSR MarkersWan P*, Zeng CW, Zhao B, Wu BM, Pu SJ and Jin WLCollege of Plant Science and Technology, Beijing University of Agriculture, Beijing 102206, P. R. China* Email: pingwan3@yahoo.com.cnAzuki bean (Vigna angularis) originated in China, and is one of Chinese most ancient cultivated crops. Geneticdiversity and DNA fingerprints of 145 accessions including 24 wild, 24 weedy and 97 cultivated azuki beanswere analyzed using SSR markers. 196 SSR markers for azuki bean, 21 from mung bean were screened todetect polymorphism, 77 SSR primer pairs revealed clear polymorphism. 165 polymorphic bands were obtainedin 145 accessions using maximally polymorphic primer pairs selected from the 77 clearly polymorphic SSRmarkers. Each primer pair detected polymorphic bands ranged from 2 to 19 with an average of 8.2 bands. Themean of Polymorphism information content(PIC)was 0.6922. The polymorphic frequency of bands amplifiedby 18 azuki bean and 2 mung bean SSR primer pairs was 38.3% and 9.5% respectively.145 accessions were divided into 13 subgroups in cluster analysis. Wild and weedy azuki beans of Japan andSouth Kroea have the closest genetic relationship. Wild azuki beans from China are closely relative with 158
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Burma’s wild. There are abundant genetic variations in Chinese landraces. Weedy azuki bean is closer geneticrelationship to the wild than to landraces and commercial cultivars. Genetic distances are 0.7871, 0.6269, and0.5389 between wild, weedy, landrace and commercial cultivars respectively. Integrated analysis of geneticdiversity on all parameters showed that wild > weedy > landrace > commercial variety. Commercial cultivarshave narrow genetic background.Landraces and commercial cultivars were classified into 14 groups on the basis of geographical origin in clusteranalysis. Cultivated azuki beans from Heilongjiang, Shanxi, Beijing and Japan have relatively higher geneticdiversity. Cultivated azuki beans of Northeast China and Japan have the closest genetic relationship. Landracesof Sichuan are clustered into a special subgroup and indicate further genetic relationship to the accessions ofother geographical origin. Cultivated azuki beans from Jiangsu and Shanxi is closely relative. Shanxi may be aimportant spreading region and pathway for gene flows of azuki bean.The specific DNA fingerprint is detected in 5 Chinese landrace, 9 wild, 3 weedy azuki beans and 1 commercialvariety. DNA fingerprints of 47 commercial varieties were gotten and differentiated.This study is important for further analysis on evolution, origin and diffusion of azuki bean. It is very useful toefficiently evaluate and develop the germplasm resources, protect variety right, serve azuki bean improvement.S 4.17 - Genetic Diversity Analysis of CIP Resources and Chinese Potato (Solanum tuberosum L)Cultivars by AFLP MarkersWang F 1, 2, Li FD 1, 2, Wang J 1, 2, *, Zhou Y 1, 2 and Sun HH 1, 21 Institute of Biotechnology of Qinghai Academy of Agriculture and Forestry, Xining, 810016, P.R China;2 The Qinghai-Tibet Plateau Biotechnology Key Lab of Ministry of Education, Xining, 810016, P.R China.E-mail: qhwf324@163.comTo apply the International Potato Center (CIP) resources in Chinese potato breeding, this study analyzed thegenetic relationship of 64 potato cultivars by AFLP marker. 12 of 81 primer combinations were screened outbased on the high polymorphism, and used these primers for analyzing the examined materials. The number ofbands of each AFLP primer pair produced was between 33 and 56 with an average of 42. 12 primer pairsamplified a total of 505 reliable bands, of which polymorphic bands were 388. The genetic distance of 64 potatocultivars was between 0.0412371 and 0.3917326. The cluster analysis showed that domestic cultivars wereclustered in one group at the genetic coefficient of 0.82 and all tested potato cultivars were clustered together atthe genetic similarity coefficient (GS) of 0.68. Shannon’s Information Index of materials was between 0.2456and 0.4988 with an average of 0.3745, Nei’s Gene Diversity Index of materials varied from 0.1510 to 0.3256with a mean of 0.2359, which indicated that genetic basis of domestic cultivars is narrow, and the geneticaccession of potato resources derived from CIP and domestic cultivars was distinct. The CIP germplasmresources will be utilized for broadening the genetic basis of Chinese potato.S 4.18 - Identifying neutral allele San of pollen-sterility in the Single Segment Substitution Lines in Riceby Using Gaozhou Common Wild Rice (Oryza rufipogon) as Donor ParentWang L, Li JQ,Wang L, Zhao XJ, Zeng RZ,M.Q. Shahid, Liu XD*, Lu YG*Guangdong Provincial Key Laboratory of Plant Molecular Breeding, South China Agricultural University,Guangzhou 510642, China. 159
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京* Email: xdliu@scau.edu.cn; yglu@scau.edu.cnPollen sterility is commonly found in the intra-specific hybrids of indica and japonica rice, which is one of themain constrains for the utilization of powerful heterosis between indica and japonica. Previous studies revealedthat at least six loci (Sa, Sb, Sc, Sd, Se and Sf) controlled the F1 pollen sterility through genes interaction. Neutralalleles at each locus have potential to overcome the F1 pollen sterility associated with each locus. Therefore,exploitation and utilization of neutral alleles are of significant importance. In this study, single segmentsubstitution lines (SSSLs) for F1 pollen sterility Sa gene locus were firstly developed by using Huajingxian74 asrecipient parent and O. rufipogon indigenous to Gaozhou as donor parent through multiple backcrossing andmicrosatellite marker-assisted selection (MAS). Then, crosses were made by using Taichung 65 (with thegenotype of SajSaj, E1) and its near-isogenic line (with the genotype of SaiSai, E4) as male parents and the SSSLsfor F1 pollen sterility Sa gene locus as female parents. The pollen fertility and spikelet fertility of F1s wereexamined in in order to identify the neutral gene for Sa locus, and the genotypes were detected at the same time.The results showed that the pollen and spikelet fertility of the two F1s between the SSSL (GZW0006 as donorparent) and E1 or E4 were 83.69±1.67%, 86.11±1.85%, 70.23±2.45% and 71.49±2.60%, respectively. Thefertility was higher than that of the control, and showed non-significant difference by t-test, suggesting that noallelic interaction exists at the Sa locus between the alleles of GZW006 and E1 or E4. Further genotype detectionfound that GZW006 and the SSSL had the same genotype as the compatibility gene San identified by previousstudy, i.e. SaM+/ SaM+// SaF- /SaF-. Evidentially, GZW006 carried the neutral gene (named SanSan) at the Sa locus,which provides valuable theoretical basis and resources for further studying and overcoming the sterility ofindica-japonica hybrids.S 4.19 - Genetic diversity of autotetraploid rice and their DNA fingerprintingWu JW, Guo HB, Chen WD, Chen ZX, Shahid M.S., Liu XD*College of Agriculture,South China Agricultural University, Guangzhou 510642,China;* Email : xdliu@scau.edu.cnAutotetraploid rice is a new germplasm developed by chromosome doubling of diploid rice. Inter-subspecificautotetraploid rice hybrids possess stronger yield potential and greater adaptability. However, lower fertility ofautotetraploid hybrids limited their application in rice production. More than 90 autotetraploid rice lines weredeveloped in our laboratory. Our objective was to analyze the genetic diversity, and to construct the DNAfingerprinting for autotetraploid rice by using SSR markers. The results showed: (1) stable polymorphism couldbe found in different lines with 99 SSR primers, with polymorphic percentage of 55.9%. Chromosome number 4and 12 produced the highest polymorphism (66%) than other chromosomes, while chromosome number 1produced lowest polymorphism (25%). A total of 285 alleles were detected with an average of 2.87 alleles perlocus. The Shannon index (I) were ranged from 0.0896 to 1.569 (mean 0.8107) and expected heterozygosity (He)were ranged from 0.0259 to 0.7855, average for all samples was 0.4852. Mean value of PIC was 0.4176 and itranged from 0.0345 (RM559) to 0.7506 (RM527). (2) DNA fingerprints of 58 autotetraploid rice lines could bedetected with 11 pairs of SSR primers,using the 0-1 number matrix to identify autotetraploid rice lines,as wellas analysis of genetic relationship between materials. (3) Genetic diversity of agronomic traits of autotetraploidrice and the correlation between agronomic traits by phenotypic analysis showed that grains per panicle and the 160
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京total number of grains per plant had large variation,while panicle length, grain length and width had littledifference. Further cluster analysis revealed that the second sub-groups of groupⅠ and group Ⅲ comprehensivecharacters are good, providing a basis for subsequent breeding.S 4.20 - DNA Profiling and Genetic Diversity of Sugarcane Germplasms by SSR LociXu LP 1, Huang XD 1, Que YX 1, Pan YB 2 and Chen RK 11 Key Lab of Sugarcane Genetic Improvement, Ministry of Agriculture, Fujian Agriculture and ForestryUniversity, Fuzhou, 35002; P. R. China;2 USDA-ARS, Sugarcane Research Units, Houma, LA, 70360.*Email: xlpmail@yahoo.com.cnFourteen highly polymorphic microsatellite (SSR) DNA markers were used for the first time in China to studythe genetic diversity of 136 important clones, including Erianthus arundinaceus (Retz) Jeswiet, wild sugarcaneclones, exotic and modern sugarcane cultivars. A total of 389 alleles were detected. The number of alleles permarker ranged from 17 to 50, with an average of 27.8. Of which, 330 alleles were polymorphic (84.8%). Thepolymorphism information content (PIC) values of these SSR markers ranged from 0.74 to 0.89 with an averageof 0.81. Cluster analysis showed that the genetic similarity coefficients among the 136 sugarcane clonesranged from 0.75 to 0.96. At incision of 0.77, these clones were divided into three groups. Group I, Group II andGroup III contained mainly primitive cultivars of S. sinense, wild sugarcane clones, and modern sugarcanecultivars (Saccharum Complex), respectively. The results of principal component analysis showed that 136clones also could be divided into three relatively separate groups. The genetic distance between Group I andGroup II was greater than between Group II and Group III. Two SSR markers, namely SMC119CG andSMC31CUQ, produced a total of 67 alleles. Both DNA fingerprints and digital fingerprints were constructed forthese 136 clones based on these two SSR loci. According to different Chinese sugarcane breeding institutes, allmodern sugarcane cultivars in China belong to one of eight series, namely, Yuetang, Guitang, Yacheng, Funong,Yunzhe, ROC, CP, and HoCP series that have different levels of genetic diversity. The Yacheng series was therichest with a polymorphism rate of 58.4 % while the HoCP series was the lowest of 38.3%. On the other hand,based on cultivar release period, the modern sugarcane cultivars tested in this study could be divided into 7series, namely, 1956-1980, 1981-1985, 1986-1990, 1991-1995, 1996-2000 and 2001-2005, respectively. Ourstudy showed that cultivars released during 1996-2000 had the highest polymorphism rate of 64.3% and thelowest rate (39.6%) was observed in the series of cultivars released during 1981-1985.S 4.21 - Hereditary Analysis of Dwarf Characteristics and Molecular Marker of Related gene in ActinidiaChinensis PlanchXu XB 1 , Jiang CY 1,2 , Liao J 1, Gu QQ 1 , Huang CH 1 and Zhang L 31 College of Agronomy, Jiangxi Agricultural University, Nanchang, Jiangxi 330045 China;2 Research Center of Fruit Tree Engineering Technology in Guizhou Province, Guiyang, Guizhou 550025 China;3 College of Gardening and Landscape, Jiangxi Agricultural University, Nanchang, Jiangxi 330045 China. *Email: xiaobiaoxu@hotmail.comThe dwarfish of kiwifruit (Actinidia spp.) is an important economic character, and one of the breeding 161
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京objectives is to obtain dwarf variety in modern kiwifruit production. Understanding of inheritance for dwarfcharacteristics will able to provide a theoretical basis for improvement of kiwifruit germplasm. The EST-SSRmarkers related to dwarf gene in Actinidia Chinensis Planch were analyzed by using sexual progenies asexperimental materials.Analyzing the separated proportion of the dwarf phenotypes in Fl individuals, we foundthat the dwarf characteristics in Actinidia Chinensis Planch was controlled by single dominant gene and namedas Ad.The separating progenies from the cross between dwarf cultivar Actinidia Chinensis Planch ‘Ganmi 5’and normal cultivar ‘Fengxiong 2’ were employed to identify the molecular markers linked to dwarf gene ofActinidia by using high effective simple sequence repeats (SSRs) marker based on expressed sequence tags(ESTs) combining with the method of bulked segregation analysis (BSA) on the automated fluorescent-labeledGenetic Analyzer system (ABI3130) with 85 pairs of fluorescent EST-SSR primers designed by ourselves. Theresults showed that the fluorescent primer pair EST-Ad042 (F:5GTTAATTTGATCGGGATGG-3; R5-GAGGAGCTTGAGCTGCTAT-3 ) could amplify specific segment of 285 bp in the dwarf pool and femaleparents, but did not detected the specific segment in the normal DNA pool (Fig.1-N) and male parents.Further research indicated that the 46 dwarf and normal individuals were obtained In the F1 progenies at randomrespectively, which were amplified screened by EST-Ad042 primers. The results indicated that the specificsegment of 285 bp was detected in all dwarf individuals, but did not found the specific segment in most normalindividuals, with the exception of the normal individuals (PT49、PT54、PT62、PT63、PT90) detected the specificsegment of 285 bp, which showed the special segment generated by gene change and recombination type onmarkers and characters in 5 normal individuals. The marker, which was dominative on the dwarf phenotype andrecessive on the non-dwarf phenotype, was obtained 94.6% of right prediction rate in F1 individuals. It was thusclear that primer of fluorescent EST-Ad042 could amplify the special EST-SSR marker in the dwarf individuals,which the molecular marker tightly linked to the dwarf genes in the Actinidia Chinenisis Planch, which could beused to identify dwarf or normal plants. The further genetic linkage analysis showed the genetic distancebetween the EST-SSR marker and dwarf gene was 8.8 cM. The specific marker of Actinidia not only could beeffective in marker-assisted selection for the dwarf character, but also provide a solid basis for the dwarf genemapping and cloning in Actinidia breeding program.S 4.22 - An intergeneric crossing of Bambusa multiplex × Dendrocalamus latiflorus and hybrididentification by morphological and molecular analysisYuan JL, Yue JJ, Ma NX, Chen YT, Gu XPResearch Institute of Subtropical Forestry, Chinese Academy of Forestry, Fuyang 311400, Zhejiang, China.E-mail: yjl_369@sina.comBambusa multiplex, a caespitose bamboo species with wide distribution in Asia, Africa, Australasia, southAmerica, etc, is evidently popular in China on account of its relatively cold-resistance, especially in Chinesetemperate regain where other sympodial bamboo resources were scarce for winter killing, which shares goodfibre for paper making simultaneously.Dendrocalamus latiflorus, the other sympodial bamboo species with giant culms up to 25 m, is a both timberand shoot-used bamboo species which grows in Asia-temperate and Asia-tropical, however, its cultivation wasrestricted in southern China by an obstacle of winter killing.Intergeneric crossing of B.multiplex × D.latiflorus were conducted with an aim to combine the cold-hardiness of 162
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京B.multiplex and high yield of D.latiflorus. Selfing of B.multiplex and D.latiflorus were developedsimultaneously for a morphological contrast of hybrids since the difference of infants and adults, and theparental death after flowering and fruit-bearing.10 putative hybrids were selected at four months’ age by a phenotypic contrasting to paternal selfed progenies,which present an evidently large leaf than that of the maternal selfed progenies. 10 putative hybrids demonstratean intermediate traits between the parental selfed progenies in leaf length, leaf width, diameter, leaf veins, leafsheath, leaf auricle, etc..AFLP molecular markers were used to confirm the hybridity. 8 pairs of primer combinations were selected forgenetic analysis of parental and 10 putative hybrids samples. All of those 10 putative hybrids consist paternalpeculiar loci, with an range of 21.97%-28.10%; Maternal peculiar loci account for 15.92%-18.70%;Non-parental loci emerge with 1.88%-4.05%.Those hybrids exhibit vigorous growth performance currently in field trial and give a maximum 3.3 cm diameterand 4.0 m height, 33 cm leaf length and 6.0 cm leaf width at fifteen month’s age, which made it possible fornext elite selection after field trial. More crossing should be conducted for genetic improvement and geneticanalysis.S 4.23 - Study on the genetic basis of high seeds per silique Brassica napus line ‘Y106’Zhang LW, Yang GS*, Liu PW, Hong DF, Li SP, Chen L and He QBNational Key Lab of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan 430070, P.R.China.* Email : gsyang@mail.hzau.edu.cnSeeds per silique (SS) is one of the three important components of yield in rapeseed and has always receivedmuch attention. In the breeding practice, Rapeseed Laboratory of Huazhong Agricultural University has foundone high combining ability and high-SS line ‘Y106’, and its SS varied from 25.9 to 30.0 in differentenvironments. For elucidating the genetic basis of high-SS, ‘Y106’ was crossed with three low to moderate SSmaterials, ‘HZ396’, ‘HZ165’ and ‘HZ168’. The heterosis and cytoplasmic effects of SS was investigated in theF1 and reciprocal F1 generations of the three crosses. SS was found to be under the control of nuclear genesinstead of cytoplasmic genes and the high-SS genotype was almost completely dominant over the low-SSgenotype. To dissect quantitative trait loci (QTL) for SS, a linkage map comprising 150 Simple Sequence Repeatand 195 Amplified Fragment Length Polymorphism markers covering 1759.6 cM in a doubled haploid (DH)population from a cross between ‘HZ396’ and ‘Y106’ was constructed. In field experiments across three seasonsand two locations in China, 140 DH lines and their corresponding parents were evaluated for SS. Usinginclusive composite interval mapping, 6 QTL for SS were identified. Of them, 3 QTL allele originated from‘Y106’ were mapped on linkage groups N8, N17 and N19 and explained 20.08%, 6.48% and 9.04% of the meanphenotypic variation respectively. These findings will facilitate illustrating the genetic basis of high-SS ‘Y106’as well as breeding strategies for SS improvement.S 4.24 - The Cross Breeding and Genetic Analysis of Hybrids of alfalfa and yellow alfalfa using SRAPmarkersZhang Y 1, Yu LQ 2, Ci ZL 3* and Gao X 1 163
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京1 College of Ecology and Environmental Science, Inner Mongolia Agricultural University, Hohhot 010019,China;2 Grassland Research Institute of Chinese Academy of Agricultural Sciences/ Key Laboratory of GrasslandResources and Ecology, Hohhot 010010, China;3 Forestry College ,Inner Mongolia Agricultural University, Hohhot 010019, China.*Email: cizl@163.comUsing sequence related amplified polymorphism (SRAP) molecular markers, 46 samples containing 31 parentalfalfa and their progenies were analyzed to study polymorphism and heterosis. Twenty four pairs produced 287polymorphic bands, averaging 11.96 bands per primer pair. The average percentage of polymorphic bands was89.69%. Samples had relatively high molecular genetic polymorphisms so it was feasible to identify the parents.Then the use of SRAP markers for prediction of heterosis in their progenies.S 4.25 - Creation and identification of wheat-barley 2H recombination materials with Isa-H1Zou HD, Wu Y , Liu HK , Shan XH , Chen X and Yuan YP *College of Plant Science, Division of Agronomy, Jilin University, 5333 Xian street,Changchun, 130062, China.* Email:yapingyuan@hotmail.comThe bifunctional α-amylase/subtilisin inhibitor(BASI) encoded by Isa-H1 on the barley 2HL chromosome caninhibit wheat α-amylase activity thereby reducing sprouting and improving quality of wheat. regenerated plantswere derived from immature embryo culture of hybrids of Zhengzhou9023 、 CB037 、 Zhongmai16 ×wheat-barley 2H alien addition lines 2H(A)、2H(B),after callus induction,subculture,differentiation andstrong seedling.screened the tissue culture seedling and their descendants using SSR molecular markers ofbarley second homology group for wheat-barley 2H recombination materials with Isa-H1.detected therecombination type of these materials respectively using genome in situ hybridzition that barley genomic DNAas probe,CS genomic DNA for blocking.eventually homozygous ditelosomic alien substitution and homozygoustransloction with Isa-H1 gene were identified.According to relatives homology between plant genome sequenceof the existence of section collinearity, using wheat EST databases, developed the second homology group ofEST-PCR molecular markers to determine the recombination of the barley chromosome segment size, locationand homoeologous group.by measuring the quality parametres of common wheat Chinese Spring (CS) , barley(Betzes) and wheat-barley 2H chromosome addition and three substitution lines (2H/2A , 2H/2B , 2H/2D),results showed that the carrying Isa-H1 gene 2H chromosome improved wheat starch, protein quality.S 5.01 - Development of High Throughput Approaches to Understand Molecular Basis of Heterosis inRice for Precision BreedingBaranwal V1, Priya M1, Umate P1, Khurana J1, Zehr U3, Tyagi A2, Kapoor S1, Mikkilineni V31 University of Delhi South Campus;2 National Institute for Plant Genome Research;3 Maharashtra Hybrid Seeds Company Limited.* Email: venugopal.mikkilineni@mahyco.com 164
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京The current trend of India’s population growth is at 1.8% per annum, suggesting that India would requiresubstantial enhancement (~50%) in rice production within a period of three decades. Hybrid rice technology hasthe potential to enhance yields to meet future food security needs. An understanding of the molecular basis ofheterosis and classification of parental germplasm based on the molecular markers pre-selected for theirheterotic potential can significantly bring down the time and cost required to develop superior hybrids withdesirable traits. Several theories have been proposed to explain the phenomenon of heterosis such as,Dominance hypothesis, Over-Dominance hypothesis, and Epistatic interactions. To develop an understanding ofthe molecular basis of heterosis, microarray-based transcriptome profiling of a Mahyco rice hybrid and thecorresponding female and male parent lines was carried out. By analyzing the transcriptome profiles of acommercial rice hybrid and its parents at eleven stages of vegetative and reproductive development,genome-wide transcript-level perturbations at single-gene resolution have been identified. Stages with highernumber of differentially expressed genes also had significantly higher proportion of genes showing non-additiveoverdominant expression in hybrids higher than in both the parents and underdominant (expression in hybridslower than in both the parents) expression profiles. Further to validate the findings, microarray data will be usedfor molecular marker development and marker-heterotic trait associations will be established in a bi-parentalpopulation. The information generated can be used to develop a database of markers correlated to heterosis,which can further be used in the development of a chip-based diagnostic/assay system to precisely select theparents for superior Rice hybrid development.S 5.02 - Development of Single Nucleotide Polymorphisms (SNPs) Detection Platforms for GeneticAnalyses and Molecular Breeding of RiceChen HDa, 1, He Ha, Zou YJ a, Chen Wa, Yu RBa, Yang Ya, Gao YMb, Xu JLb, Zhou SCc, Li Ya, Li ZKb, and DengXWaa College of Life Sciences, Peking University, Beijing 100871, China.b Chinese Academy of Agricultural Sciences, Beijing 100081, China.c Guangdong Academy of Agricultural Sciences, Guangdong 510640, China.* E-mail: chenhaodong@gmail.comMolecular markers are used more and more in rice breeding. Single nucleotide polymorphisms (SNPs) show alot of advantages comparing to other markers, which can be used widely in high-density genetic mapsconstruction, association mapping and marker assisted selection. Here, around 132,000 SNPs meeting thecriterion for our SNPs detection assays were selected by comparing the genome sequences of several ricevarieties including Minghui 63, Zhenshan 97, PA64S, 9311 and Nipponbare. 3090 SNPs distributed evenlyalong the rice whole genome were selected and confirmed by PCR and sequencing, and most of them wereconsidered to be suitable for both Illumina GoldenGate assay and optical thin-film biosensor chips, twoplatforms we have generated. A set of 384 SNPs were further selected to analyze around 350 global ricebreeding varieties and a pedigree of elite variety Huanghuazhan. The phylogenetic tree revealed several distinctgroups, and the pedigree analysis showed interesting conserved and changed sites along the rice genome duringthe breeding process. These platforms provide a foundation for further diversity analysis, mapping andmarker-assisted selection in rice research. 165
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京S 5.03 - Development of InDel markers for Low glutelin content1 (Lgc1) gene in rice (Oryza sativa L.)Chen T, Tian MX, Zhang YD, Zhu Z, Zhao L, Zhao QY, Lin J, Zhou LH, Wang CL*Institute of Food Crops, Jiangsu Academy of Agricultural Sciences, Jiangsu High Quality Rice R&D Center,Nanjing Branch of China National Center for Rice Improvement, Nanjing 210014, China.* Email: clwang@jaas.ac.cnRice with low glutelin content is quite effective as a kind of functional food for patients with kidney disease.With an increasing number of people in this disease, there has been a great demand for this kind of rice ininternational market in recent years, so it is very necessary to develop commercial varieties of low glutelincontent. Lgc1, as an excellent genetic resource with low glutelin content, has been given more and moreattention by breeders. The mutation mechanism of Lgc1 was elucidated to be a 3.5kb deletion between twohighly similar glutelin genes GluB4 and GluB5, which located on chromosome 2S and several markers tightlylinked with Lgc1 were reported for genotyping selection. However, these markers always showed nopolymorphism in breeding population and couldn’t be applied in practice. To improve the precision ofmarker-assisted selection for low glutelin-content gene Lgc1 in breeding, we developed four pairs of InDelmarkers based on a 3.5kb deletion in this study. In theory, InDel-Lgc1-1 and InDel-Lgc1-A were co-dominantmarkers, but special Taq DNA polymerase for amplifying large fragment DNA sequences must be used and itwould increase the test cost and reduce the possibility in practical application. To distinguish genotypes of Lgc1more effectively, another markers designated as InDel-Lgc1-2 and InDel-Lgc1-B were synthesized ascomplements for InDel-Lgc1-1 and InDel-Lgc1-A. The double PCR detections indicated that three differentgenotypes of Lgc1 could be clearly distinguished by adding InDel-Lgc1-1 and InDel-Lgc1-2, InDel-Lgc1-A andInDel-Lgc1-B into a single reaction tube, respectively, which were completely consistent with the results of theprotein analysis. Thus, as a simple and low-cost technique, these InDel markers could be used widely to identifydifferent varieties with Lgc1 gene and apply in marker-assisted selection of low glutelin-content rice.S 5.04-Genome-wide Responses to Selection and Genetic Networks Underlying Submergence andDrought Tolerances in Rice (Oryza sativa L.)Fu BY 1,2, Ali AJ2, Gao Y M 1,2, Xu JL 1,2, Wang WS 1,2, Zhang F 1, Ismail A 2, Zheng TQ 1, Liu S H 1, DomingoJR 2, Jiang YZ 1,2, Yu SB 3, Vijayakumar CHM, Zhao MF2, Zhu LH 1, Mackill D 2 and Li ZK 1,21 The Chinese Academy of Agricultural Sciences, Beijing 100081, China;2 International Rice Research Institute, DAPO Box 7777, Metro Manila, Philippines; 3 National Key Laboratoryfor Crop Improvement, Huazhong Agricultural University, Wuhan 430070, China.* Email : J.Ali@cgiar.orgSubmergence and drought are two important but contrasting factors limiting rice yield and stability that mayoccur in different times of the same crop season in many rainfed areas of Asia. Empirical data suggest that ricetolerances to submergence (ST) and drought (DT) are negatively correlated. To facilitate the geneticimprovement of both ST and DT in rice, the genetic overlap of ST and DT of rice were genetically dissectedusing DNA markers and over 300 introgression lines (ILs) selected for either ST or DT from 20+ BCpopulations. Development of elite rice lines tolerant to both submergence and drought involved a large scalebackcross breeding effort was initiated in 1998 using IR64, Teqing and a new plant type (NPT) line as recurrent 166
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京parents and 64 diverse parents as donors. A total of 264 NPT, IR64 and Teqing BC2F2 populations were screenedunder submerged field conditions for 15 days at seedling phase. We selected 1,585 plants that survived the stress.Of these, 72 BC2F3 and 83 BC3F2 lines from 9 crosses between the recurrent parents and three donors, (TKM9,Khazar and FR13A) were used as materials to identify genes/QTLs controlling ST in rice. These BC progenieswere genotyped with approximately 100 anchor simple sequence repeats (SSR) markers and evaluated in areplicated experiment of submergence to identify the introgressed donor segments in the ST recipients. Linkagedisequilibrium analyses based on marker genotypic data and the survival frequency of the BC progeny wereconducted to identify the introgressed donor genomic regions that potentially harbored gene(s) associated withST.S 5.05 - QTL detection of concentration of micro-nutrients and metal elements in RiceHu X 1, Jia Q 1, Xu Q 1, Shi YM 2, Sun Y 1, Zhu LH 1, Xu JL1*, Li ZK 1,31 Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081, China;2 Institute of Rice, Guangxi Academy of Agricultural Sciences, Nanning, 530007, China;3 International Rice Research Institute, DAPO Box7777, Manila, Philippines.* Email: xujlcaas@yahoo.com.cnIron, zinc and selenium are important micro-nutrients to human health. Malnutrition of micro-nutrients is aserious problem associated with resource poor population in many developing countries. Regarding high costand technical difficulty in testing of micro-nutrients, QTL identification and marker-assisted pyramiding ofmultiple favorable QTLs are considered as a practical strategy to develop micro-nutrients dense variety. In thisstudy, two BC1F8 line populations were used for QTL mapping for Fe, Zn, Se, which were derived from the twocrosses between two elite indica varieties, Ce258 and Zhongguangxiang1 (ZGX1) as recurrent parents and ajaponica breeding line, IR69428-6-1-1-3-3 (IR69428) from IRRI as the donor parent. The donor parent IR69428had twice concentration of Fe and Zn than those of the recurrent parents, Ce258 and ZGX1. Correlation analysisindicated that highly significant positive correlation existed between Fe and Zn, and between Fe and Se but nosignificant correlation between Zn and Se. Based on 147 SSR genotyping data, QTL mapping for concentrationof Fe, Zn and Se were performed using single marker analysis by SAS program. Total 14 (4 for Fe, 5 for Zn and5 for Se) and 12 (5 for Fe, 4 for Zn and 3 for Se) QTLs were identified in Ce258/IR69428 and ZGX1/IR69428BC1F8 populations, respectively. The favorable alleles increasing concentration at all loci were from t124 in theCe258/t124 population, while the alleles increasing concentration at half loci were from ZGX1 in theZGX1/t124 population. Almost no a common QTL for the same trait was detected in the two backgrounds,indicating genetic background had a large effect on QTL expression of micro-nutrients. Three important regionsi.e. RM565-RM570 on chromosome 3, RM469-RM225 and RM258-RM340 on chromosome 6, RM277-RM511on chromosome 12 affecting Fe, Zn or Se with favorable alleles from t124 were candidate regions formarker-assisted selection. Sister introgression lines with high Fe, Zn or Se from the same population werecrossed and developed F2 populations for QTL verification and marker-assisted pyramiding to select high yieldwith high nutrition.S 5.06 - Genetic Diversity in Inbred and Hybrid Rice GenotypesIslam M R 1, Yasmin F 2, Rehana S 2, Hossain Z 3 and Salam MA 1 167
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京1 Plant Breeding Division, BRRI, Gazipur;2 Department of Biotechnology, Khulna University, Khulna;3 Variety testing wing, Seed Certification Agency, BangladeshThe study was conducted to assess the genetic diversity among the inbred and hybrid rice varieties using simplesequence repeat (SSR) markers. The research was carried out at Plant Breeding division, Bangladesh RiceResearch Institute (BRRI), Joydebpur, Gazipur, Bangladesh. About one hundred SSR markers were used tocharacterize 24 rice genotypes. These genotypes were comprised of four salt tolerant, one tidal submergencetolerant with other nine high yielding inbred, developed by BRRI and International Rice Research Institute(IRRI). Ten hybrid rice varieties originated from China were also included. Among the used SSR markerssixty-one (61) were found polymorphic. The highest number of allele (23) was found for RM6959 and thelowest number of allele (5) was found for the RM559, RM289 and RM8300. The polymorphism informationcontent (PIC) value was ranged from 0.6008 (RM8300) to 0.9267 (RM495), with an average of 0.8563.Frequency of the major allele ranged from 8.7% (RM495) to 52.17% (RM8300), with an average of 22.66%.Based on the Nei’s (1983) genetic distance matrix the genetic distance was 100% to 53.52%. Dendogram wasdrawn by using the unweighted pair group method with arithmetic means (UPGMA) and two main distinctclusters were identified. A cluster was consisted with mostly inbred genotypes while the second cluster hadmostly hybrids. This scientific information could be used for further improvement of rice genotypes throughappropriate breeding program.S 5.07 - Genetic and Physico-Chemical Study of Aromatic Rice Germplasm Through Molecular MarkersJewel ZA1, Patwary AK 2, Begum SN 31,2 Department of Genetics and Plant Breeding, Bangladesh Agricultural University, Mymensingh-2202,Bangladesh.3 Plant Breeding Division, Bangladesh Institute of Nuclear Agriculture, Mymensingh-2200, Bangladesh.For selection of aromatic rice lines twenty six rice genotypes were used to evaluate agronomic characteristics,aroma detection through sensory test and genotypic analysis using SSR markers. Phenotyping for grain quality,yield attribute data were performed after harvesting the grain at the experimental field of Plant BreedingDivision of Bangladesh Institute of Nuclear Agriculture (BINA), Mymensingh. Aroma was detected by 1.7%KOH as a sensory test. For aroma, six rice genotypes were detected as strong aroma; 7 for moderate aroma; 10for slight aroma and three for no aroma. In case of grain shape study, 24 genotypes were evaluated as slenderand 2 as medium. In this study, aroma had significant and positive association with grain length width ratio;significant and negative association with grain width, significant and negative association gelatinizationtemperature, and not significant with grain length. Gelatinization temperature had non-significant and negativecorrelation with grain length, significant and negative association with grain length width ratio, significant andpositive association grain width. Grain length had significant and negative correlation with grain width;significant and positive correlation with length width ratio. Grain width had significant and negative correlationwith length width ratio. Three SSR primers viz; RM223, RM515 and RM342 were used for identifying fgr genelocus in 26 rice genotypes. The primer RM223 identified the fgr locus as homozygous condition for 6 as strongaromatic, 7 moderate aromatic, 10 slight aromatic and the rest 3 as non aromatic. The primer RM515 detected 4 168
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京as strong aromatic, 6 as moderate aromatic, and 16 as slight to non aromatic. The primer RM342 detected 3 asstrong aromatic, four as moderate aromatic, 19 were slight to non aromatic. Compare among the three markers, -RM223 detected the highest number of fgr locus in aromatic rice genotypes. Finally, it can be said that amongthe three primers, RM223 responded best in all the 26 rice genotypes because RM223 primer could be able toidentify aromatic and non-aromatic genes having higher yield with good agronomic performance and othergrain quality traits. These elite lines could be readily used in breeding programme for release aromatic ricevariety with considerable yield.S 5.08 - Fine mapping of the region on rice chromosome 7 controlling spikelets per panicle and yieldcomponents from Oryza minuta to cultivated rice, O. sativa L.Le-Hung Linh1, Nguyen-Thi Hang2, Ju-Won Kang3 and Sang-Nag Ahn3*1 Molecular Biology Department, Agricultural Genetics Institute, Tu Liem, Hanoi, Vietnam;2 Agro-biotechnology Department, Food Crops Research Institute, Thanh tri Hanoi, Vietnam;3 Department of Agronomy, College of Agriculture & Life Sciences, Chungnam National University, Daejeon305-764, KoreaNear isogenic line (NIL) IL-34 developed by introgressing chromosomal segment from an accession of Oryzaminuta (2n=48, BBCC, Acc. No. 101141) into the O. sativa subsp. japonica cv. Hwaseongbyeo, showedsignificantly higher number of spikelets per panicle (SPP) than the recurrent parent Hwaseongbyeo.Quantitative trait locus (QTL) analysis in the F2 generation derived from the cross between IL-34 andHwaseongbyeo revealed that spp7, a QTL was located in the pericentromeric region of chromosome 7. Thedistribution of spikelets per panicle followed 3:1 ratio for single locus segregation. The additive effect of the O.minuta allele at the spp7 QTL was 23 spikelets per panicle and 43.6% of the phenotypic variance could beexplained by the segregation of the SSR marker RM21596. To fine-map the spp7 as a step for map-basedcloning, we carried out fine-scale mapping with 3,700 F2 plants derived from the cross between IL-34 andHwaseongbyeo, and detailed quantitative evaluation of main panicle traits and grain yield was performed. Onehundred and eighty-nine F2 plants having informative recombination breakpoints within the region flanked bytwo SSR markers RM500 and RM21615 were identified and used for fine mapping of spp7. spp7 was mappedbetween the SSR markers RM21596 and RM418 which was approximately 441-kb in length based on thephysical map of the region. Of great interest, the QTL region also had effects on primary branch number (PB),grains per panicle (SP) and grain yield (YD). These results are very useful for transferring or pyramiding spp7by molecular marker assisted selection in rice breeding programs and for cloning spp7 by map-base cloning.S 5.09 - Differential global genomic changes in response to different drought stress in two rice cultivarsrootMa TC1,2,3 *, Chen RJ3, Yu RR1,3, Zeng HL2, Zhang DP21 Rice Research Institute; Anhui Academy of Agricultural Sciences, Key Laboratory of Rice Genetics andBreeding of Anhui Province Hefei, Branch of National Rice Improvement Center, Hefei 230031;2 National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan 430070;3 Institute of Subtropical Agriculture, The Chinese Academy of Sciences, Changsha 410125.E-mail:tingchenm@163.com 169
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Drought stress is one of the most severe abiotic stresses that can injure plant life and development. Worldwide,one-third of the total land area is continually stressed by drought, thereby causing substantial crop losses inagricultural production. In order to improve plant resistance to drought, agricultural scientists have beenworking on transgenic technology and traditional breeding methods in response to this challenge.We used the Affymetrix rice genome array to study the gene expression change in response to low (T1 treatment),middle (T2 treatment), and high osmotic stresses (T3 treatment) in rice root of two different cultivars(Xingfengzao119 and Aihua5) at the transcription level. Our experiment results as followed:The differences of response ways to drought between Xingfengzao119 and Aihua5 under T1 treatment were thatoxygen and the genes activities of electron transport, xylan, malic acid and chitin metabolism, protein folding,antibiotics biological synthesis and transcriptional regulation were lower that those of Aihua5. And the activitiesof genes encoding endogenous response and starch metabolism were higher than those of Aihua5. Thedifferences of response ways to drought between Xingfengzao119 and Aihua5 under T1 treatment were similarto T1 treatment, but the activities of gene encoding a histone deacetylase and arginine, polyamines biologicalsynthesis were significantly lower than those of Aihua5 and the activities of gene encoding jasmonate acid andABA biological synthesis were significantly higher than those of Aihua5. The differences of response ways todrought between Xingfengzao119 and Aihua5 under T3 treatment were that the activities of gene encoding GTPregulation, transcriptional regulation, electron transport and photorespiration were significantly lower than thoseof Aihua5 and the activities of gene encoding carbohydrate metabolism, protein folding and transport, and so on,were significantly higher than those of Aihua5.S 5.10 - Genetic Diversity in Inbred and Hybrid Rice GenotypesIslam MR 1, Yasmin F 2, Rehana S 2, Hossain Z 3 and Salam MA 41 Plant Breeding Division, BRRI; 2Department of Biotechnology, Khulna University; 3 Seed CertificationAgency; 4BRAC BangladeshGenetic diversity was assessed for 24 rice genotypes composed of 14 inbred and 10 hybrid varieties using 100SSR (simple sequence repeat) markers. The research was carried out at Plant Breeding division, BangladeshRice Research Institute (BRRI), Gazipur. The inbred varieties were comprised of five landraces (four salttolerant and one tidal submergence tolerant) and nine high yielding varieties developed by BRRI and IRRI(International Rice Research Institute). The hybrid rice varieties were originated from China. Among the SSRmarkers 61 were found polymorphic. The highest numbers of allele (23) were found for RM6959 and the lowestnumbers (5) for the RM559, RM289 and RM8300. The polymorphism information content (PIC) values rangedfrom 0.6008 (RM8300) to 0.9267 (RM495) with an average of 0.8563. Frequency of the major alleles rangedfrom 8.70% (RM495) to 52.17% (RM8300) with an average of 22.66%. Based on the Nei’s (1983) geneticdistance matrix, the genetic distance ranged from 53.52% to. 100% Dendogram was drawn by using theun-weighted pair group method with arithmetic means (UPGMA). The rice genotypes grouped into two distinctclusters. One consisted mostly with inbreeds while the other group mostly with hybrids. The study thusefficiently isolated the inbred rice from hybrid varieties through molecular markers. 170
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京S 5.11 Drought tolerance QTL fine mapping in riceNie YY, Liu HY, Cai YH, Luo LJ*Shanghai Agrobiological Gene Center, Shanghai 201106, China.Rice Research Institute, Jiangxi Academy of Agricultural Sciences, Nanchang 330200, China.* Email: happynyy@163.comThe improvement of drought tolerance in rice is an arduous and long-term task.QTL mapping, fine mapping andgene cloning of drought-related traits, and the realization of molecular breeding for drought tolerance, is the hotresearch in recent years. QTL mapping in rice is mainly focused on root, such as root thickness, root length, rootnumber and root distribution, as well as physiological characteristics, such as ABA, osmotic adjustment, RWCand so on, and positioned a large number of QTLs. However, further research using of these large population tocarry out fine mapping, there exists great difficulty in real work. In this research, we constructed NILs andnarrow down the interval to fine mapping in target interval in an RIL population of 520 lines from a crossbetween an indica paddy rice and upland japonica rice.Under the condition of drought stress, we survey the yield traits in the target interval of Chr2, Chr4 andChr7.The NILs has higher yield and spikelet fertility than Zhenshan97B as the recurrent parent. In 2008, TheANOVA result showed significant difference between water treatments except PL in the target population ofChr2.Highly significant positive correlation between GY and BY, then PW, HI, GNP, PN, SN, SPD and SFunder well-watered condition, while under drought stress condition, PW has highest positive correlation withGY, then GNP, HI, BY, SF, SN, SPD, PN and PL. Path analysis partitioned the correlation coefficients intodirect and indirect effects. In well-watered condition, BY had the highest direct effect on GY, followed by HI,SN, PN, PW, SF, HGW, GNP, PL and SPD. While in drought stress condition, HI had the highest direct effect onGY, followed by BY, GNP, PW, SN, PN, HGW, PL, SPD and SF. The path coefficient was not consistent withcorrelation coefficient.Quantitative trait loci (QTL) mapping was carried out for grain yield, yield components and otherdrought-tolerant relative traits in the target interval of Chr2 in rice under normal water condition and droughtstress in 2008. Based on composite interval mapping method at the threshold LOD≥2.0, under normal watercondition, total 11 main-effect QTLs were detected for PL, PN, SF, HGW, IHD, HD, PH, TN, PND, SPD andCT. While under drought stress treatment, total 9 main-effect QTLs were detected for PN, LWP, HGW, IHD,HD, BCT, TN, SPD and CT. Different place and different repeats detected different QTLs, and in the hotinterval RIO02052 ~ RM02002, several QTLs were simultaneously or adjacently identified under twocontrastive water conditions in different repeats,and find 5 putative genes in this hot points.In the target interval of Chr4, we detected 3 QTLs for PL, PN and SN. The QTL for PL was identified indifferent interval RM17308~RM17305 and RM17349~RM17190,and the contributions were 18.8% and20.42%.The QTL for PN was identified in the interval RM1354 ~ RM17308,and the contribution was11.47%.The QTL for SN was identified in the interval RM17308~RM17305, and the contribution was 28.08%.S 5.12 - QTL Mapping of Outcrossing characteristics Using a Recombinant Inbred Line of a Crossbetween Indica and Japonica Rice (Oryza sativa L.)Ren GJ*, Gao FY, Su XW, Bai G, Lu XJ, Ren JS, Li ZH, Liu WB, Jiang TG, Liu GCSichuan Academy of Agricultural Sciences, Chengdu, 610066.* Email: guangjun61@sina.com 171
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Stigma exsertion and flowering time are two important traits affecting outcrossing characteristics, which play anessential role in hybrid seed production. Stigma exsertion in rice is thought to be controlled by polygenes(Virmani and Athwal 1973, 1974), QTL mapping and associate mapping for the frequency of stigma exsertionhad been conducted (Miyata et al, 2007; Uga et al. 2003;Yamamoto et al. 2003; Yan et al, 2009; Yue et al,2009; ). In rice, genetic analyses of flowering time mostly focused on the heading date (Lin et al., 1998, 2002;Yamamoto et al., 2000; Yano et al., 1997, 2001). Little is known about the genetic basis of the starting time ofrice flowering and flowering duration at the day. The main objective of the present study was to investigate thegenetic basis of stigma exsertion, as well as that of the starting time of rice flowering and flowering duration atthe day.A recombinant inbred line (F8) population with 184 lines, derived from a cross between Chuanxiang 29B (Oryzasativa L. subsp. indica) and Lemont (Oryza sativa L. subsp. japonica), was genotyped with 98 SSR markers,and phenotyped for three traits of stigma exertion at Chengdu site and flowering number (FL) in three floweringtime (before 11:00, 11:00-12:30 and 12:30-14:00) both at Chainan and Chengdu sites.A genetic linkage map, contained 98 SSR markers, was constructed. The map covered a total of 1244.8cM withan average interval of 17.25cM.A distinctive difference of three stigma exertion between two parents was observed, and Chuanxiang29Bshowed consistently higher values than lemont. The phenotypic values of three stigma traits showed continuousvariation in RIL population. Six QTLs of stigma exertion were detected on chromosomes 2, 3 and 9. Two QTLs(qSSE-2 and qSSE-3) were associated single stigma exsertion (SSE), two (qDSE-2 and qDSE-9) with dualexsertion (DSE), and two (qTSE-2 and qTSE-3) with total exsertion (TSE). Specially, the intervalRM3316-RM1367 on chromosome 2 associated with all three exertion characteristics, and RM5489-RM5480on chromosome 3 affected not only SSE but also TSE. The Chuanxiang29B-derived allele contributed toincreasing the stigma exertion at all detected QTLs, except for those located on chromosome 2.In Hainan site, the mean FL in three flowering time of Chuangxiang29B were 35.71%, 56.63% and 7.65%,respectively, which showed consistently values in Chengdu site. However, a distinctive different was observedin the mean FL of lemont at two sites. The FL before 11:00 and in 11:00-12:30 had a significant negativecorrelation with that between 12:30-14:00 at both sites. The FL before 11:00 was positive correlated with thatbetween 11:00-12:30 at Chengdu site, in contrast to the results at Hainan site, which had no significantcorrelation. One major QTL for FL before 11:00, named qFT10-1, was detected on chromosome 10 at Chengdu,with an LOD score of 8.69 and accounted for 72.51% of the phenotype variance. Another QTL, qFT5 wasidentified on chromosome 5 at Hainan, with a minor effect on FL before 11:00. The Chuanxiang29B allelescontributed an increasing effect on FL at qFT10-1 and qFT5.These QTLs may be useful in the QTL pyramiding for making high outcrossing in rice plants which wouldincrease the seed production in hybrid rice.S 5.13 - Genetic background effects on introgression, QTL expression and cryptic genomic variation dueto subspecific differentiation in riceWang Y 1,2, Cheng LR1, Sun Y 1, Wang WS 1, Guan YS 1, Zhu LH 1, Xu JL1, *, Li ZK1,31 Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081, China;2 Shenyang Agricultural University, Shenyang 110161, China; 172
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京3 International Rice Research Institute, DAPO Box7777, Manila, Philippines.* Email: xujlcaas@yahoo.com.cnTwo large sets of reciprocal introgression lines (ILs) derived a japonica variety ‘Lemont’ and indica variety‘Teqing’, were analyzed using molecular markers to understand the transmission pattern of donor segments inreciprocal indica/japonica recipients of rice and the effects of genetic background on QTL expression of flagleaf width (LW) and spikelet number per panicle (SNP) under the two diverse environments (Beijing andHainan). Comparison of the linkage maps from the two ILs and RIL populations from the same cross revealed ahigh frequency and genomewide distribution of inverted marker orders, suggesting the widespread presence ofintraspecific cryptic genomic variation presumably due to subspecific differentiation within O. sativa. Theaverage introgression frequency of japonica genome into indica background was in agreement with the expectedvalue while that of indica genome into japonica background was much higher than the expected. Asymmetricover-introgression of the indica alleles on chromosome 7 and many specific regions on other chromosomes inthe japonica genetic background is consistent with the general trend observed in the RIL and F2 populations ofthe same cross, suggesting a high genetic load in the japonica gene pool, and japonica genome experienced avery severe bottleneck by artificial domestication. Genetic background was found to have overwhelming effectson QTL expression, as 87.3% and 79.6% of QTLs for the two traits were detected in only one of the GB inBeijing and Hainan, respectively. Many main-effect QTLs identified in the ILs had previously been detected asepistatic loci in one or more of 6 related mapping populations and their main effects were largely predictablefrom the prior epistatic effects. These results provide compelling evidence for the importance of epistasis indetermining quantitative trait variation and impel caution in applying QTL information to marker-assistedimprovement of complex traits in rice and other crop plants.S 5.14 - Introgression of Xa7, Xa21 for improving bacterial blight of a three-line hybrid rice restorer lineYihui 1577 and hybrids by molecular marker-assisted selectionXu JY, Jiang JF and Mou TM*National Key Laboratory of Crop Genetic Improvement, National Center of Plant; Gene Research (Wuhan),Huazhong Agricultural University, Wuhan 430070, China.* Email: tongmin58@mail.hzau.edu.cnYihui1577 is an elite restorer line widely used in hybrid rice production in China. The line and its derivedhybrids are susceptible to Bacterial Blight (BB) caused by Xathomonas oryzae pv. oryzae (Xoo). In this study,two resistant genes Xa7 and Xa21 were introgressed into Yihui1577 by backcrossing and marker-assistedselection. Six breeding lines, HH1202(Xa7), HH1203(Xa7), HH1204(Xa21), HH1205(Xa21),HH1206(Xa7+Xa21) and HH1207(Xa7+Xa21), and their derived hybrids were evaluated with 7 Xoo isolates(PXO61, PXO99, ZHE173, GD1358, FuJ, YN24 and HeN11) from the Philippines and China. The resultsshowed that lines containing Xa7 and their hybrids were resistant against six Xoo isolates except for PXO99.The lines containing Xa21 and their hybrids were resistant against six Xoo isolates except for FuJ. The linescontaining Xa7 and Xa21 and their hybrids were resistant against all of seven Xoo isolates. The replicatedexperiments showed that the breeding lines and their hybrids had similar agronomic traits, growth durations, 173
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京yields and rice qualities with the recipient Yihui1577 and its hybrids except from improving bacterial blightresistances.S 5.15 - Development of STS Marker Linked to Blast Resistance Gene Pi9 and Marker-Assisted SelectionYin DS, Xia MY, Li JB, Wan BL, Zha ZP, Du XS and Qi HX *Food Crops Institute, Hubei Academy of Agricultural Science, Wuhan, 430064, China;* Email:qihx@hbaas.comRice blast is the most devastating disease worldwide, which brings huge losses in rice production every year.Application of broad-spectrum and durable resistant genes is an important strategy of rice breeding. In this study,75-1-127, an indica cultivar carrying the rice blast broad-spectrum resistance gene Pi9 was used as a resistancegene donor in rice backcross breeding for genetic improvement of Yangdao 6 and R6547. By alignment ofgenomic sequences from the Pi9 locus of 75-1-127, Yangdao 6 (indica) and Nipponbare (japonica), a sequencevariation was found in the upstream region of the Pi9 gene with an interval of about 26 kb. Comparing with75-1-127, there is a 5bp deletion in Yangdao 6 and a 3bp deletion in Nipponbare, respectively. PB9-1, anco-dominant STS (sequence-tagged site) marker closely linked to Pi9, was designed and developed formolecular-assisted selection (MAS). Genotyping of the parents and segregation population using theco-dominant PB9-1 marker showed the same result as using a dominant marker pB8. Eight backcross breedinglines carrying Pi9 were obtained, among which four lines contained the genetic background of Yangdao 6 andthe other four lines had the R6547 background. Under natural disease infection conditions in Enshi County andYichang County of Hubei Province, the Pi9-carrying lines showed blast disease resistance at various levelshigher than Yangdao 6 and R6547. The result of resistance identifying also revealed that the lines derived fromR6547 showed higher resistance than the lines from Yangdao 6, and indicated that the Pi9 gene may performedvarious resistance with different genetic background. An R6547-derived rice line 08C893 was utilized as parentof rice hybrid and the hybrids showed blast disease resistance as compared to the control Yangliangyou 6. Theresults suggested PB9-1 is an effective co-dominant marker for MAS of Pi9 and in Hubei area Pi9 is a valuableresistance gene for the control of rice blast disease.S 5.16 - Developing inDel marker for molecular breeding of Xa26/Xa3 in riceZhang DCBitechnology Research Center, China Three Gorges University, Yichang, Hubei, China.* Email:zhangdc227@163.comIncorporation of resistance genes into elite rice cutivars with marker-assistance is the most effective andeconomical strategy to control rice disease in breeding programs. Bacterial blight(BB), caused by Xanthomonasoryzae pv. oryzae (Xoo), is one of the most devastating rice disease worldwide. Up to present, about thirty genesconferring resistance to bacterial blight have been identified in rice, six of them have been cloned, and anotherseven have been fine mapped. Although some DNA markers for rice bacterial blight resistance have beendeveloped, but most are not suitable for routine use in Marker-assisted selection (MAS). The BB resistance geneXa3, now known as Xa26/Xa3, which was first identified in Minghui63, has been widely used in breedingprograms and played an important role in protecting rice from BB. In this report, an inDel marker DQ4, was 174
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京developed to distinguish the domiant Xa26/Xa3 allele from the recessive allele, based on the PCR-fragmentlength difference between Xa26/Xa3 containing rice lines (IRBB3, Minghui63, Zhachanglong) and 15 cutivarssusceptible to Philippine Xoo strains PX086. PCR fragments tightly linked to Xa26/Xa3 gene were subclonedand sequenced. A 39bp DNA fragment was found to be deleted in IRBB3, Minghui63 and Zhachanglong.Because it is tightly linked with the resistance gene and easy to use through analysis of amplification products.The inDel marker is ideally suited for marker-assisted selection for Xa26/Xa3 gene and has been successfullyapplied in our rice breeding program. This result should contribute to incorporation Xa26/Xa3 gene into newrice cultivars and elite lines.S 5.17 - Putative genetic networks underlying seedling cold tolerance in rice detected by selectiveintrogressionZhang F 1, Ma XF 3, Gao YG 1 and Li ZK 1,21 Institute of Crop Sciences/National Key Facility for Crop Gene Resources & Genetic Improvement, ChineseAcademy of Agricultural Sciences, Beijing 100081, China;2 International Rice Research Institute, DAPO Box 7777, Metro Manila, Philippines;3 Institute of Rice, Liaoning Academy of Agricultural Sciences, Shenyang 100101, China.Low temperature at the seedling stage has been an important factor limiting rice production in Northeast China.Developing rice cultivars with good seedling cold tolerance (CT) is the most effective approach to solve theproblem. To efficiently discover and utilize useful genes from rice germplasm resources is the key to achievenew breakthroughs in rice improvement. In this study, C418, an elite japonica restorer (Oryza sativa L.) fromNortheast China, was used as the recurrent parent and indica variety BG300 from Sri Lanka was used as donorto obtain BC2F2 bulks by crossing and continued backcrossing. In 2002, 11(2.8%) seedlings were screened forCT under the low temperature of the field conditions (from 10℃ to 15℃) in LAAS. From 2003 to 2005,2-round pedigree selections for seedling CT following initial screening method were conducted to obtain 81 CTBC2F6 introgression lines (ILs). Replicated experiments under both normal (non-stress) and naturallow-temperature (stress) conditions indicated that BG300 had poor CT but there were transgressive segregationsin all ILs for CT improvement, indicating the presence of ‘hidden genetic variation’ for improved CT inapparently cold-susceptible donor. Performances of BC2F6 ILs selected for trait related to seedling CT undernatural low-temperature in replicated progeny testing under the controlled phytotron low-temperature conditionsshowed that a total of 20 promising ILs derived from the same BC2F2 IL which had significant higher survivalrates than C418.An attempt to identify CT QTLs based on genetic hitch-hiking theory in BC breeding population was describedin this study. The selected BC2F6 ILs were genotyped with 100 well distributed simple repeat sequence (SSR)markers and the genotype of 11 selected BC2F2 ILs were inferred based on the genotype of 81 BC2F6 ILs.Simple chi-square tests based on genetic hitch-hiking theory across the genome to detect alleles at individualloci that responded significantly to selection were conducted on the BC2F2 population. A total of 16 CT lociwere detected in which the BG300 (donor) allele and genotypic frequencies deviated significantly fromexpectation. 17 loci were confirmed by Z-tests between the donor introgression frequencies of BC2F6 sister lineswith separating CT performance from the same BC2F2 IL, including 11 loci (68.8%) detected in BC2F2population. Linkage disequilibrium (LD) results indicated the non-random associations among unlinked loci 175
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京constructing complex genetic networks following the method proposed by Li et al (submitted). Our resultsindicate that directional selection for CT aims at a group QTLs even genetic networks but not individual QTL.Backcross breeding coupled with efficient screening is a powerful way to improve CT by using the hiddendiversity of CT sensitive indica germplasm and discover valuable genetic information for pyramiding design.S 5.18 - QTL detecting for yield and its components using yield selected introgressed populations derivedfrom a key parent of hybrid riceZhang HJ 1, Qian YL 1, Wang H 1,2, Zhu LH1, Gao YM1,3 and Li ZK1,31 Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081;2 Anhui Provincial Crop Introduction and Breeding Cencer, Hefei 230601;3 International Rice Research Institute, DAPO Box 7777, Metro Manila, PhilippinesThree advanced backcross populations under selection derived from the intraspecific crosses betweenShuhui527 (Oryza sativa L. spp. indica) and its respective donors such as ZDZ057 (Oryza sativa L. spp. indica),Fuhui838(Oryza sativa L. spp. indica) and Teqing (Oryza sativa L. spp. indica) were used to detect quantitativetrait loci (QTL) associated with yield and its components. Based on the analysis of 140 simple sequencerepeat(SSR) polymorphic markers between parents distributed throughout the genome, using sing-point analysis(SPA) and X2 statistics for genotypic frequencies, 60 and 28 QTL above an experiment-wise significancethreshold (P<0.01) were identified in three populations, respectively. 46.7% QTL (that is, 28 out of 60) werecommon in at least two environments; while the beneficial alleles from donors were favorable for 50% of yieldand its component QTL including all nine yield related traits except for 1000-grain weight using SPA method.For both mapping methods, twelve QTL were simultaneously detected in three populations. In comparison withthe previous studies, sixteen out of sixty loci shared the same or similar genomic regions for nine traits exceptthe panicle number and grain yield per plant. This study mainly detected the favorable genomic regions withlarger effects on yield and its components from different donors using three selected backcross populations.Furthermore, the cluster of QTL with positive effects on correlated traits provided good strategy for developingbreeding materials, and also offers an opportunity for marker-assisted selection and pyramiding breeding.S 5.19 - Hybrid rice parents transformed with foreign genomic DNA and backcrossed into CMS rice lineBing1A with blast resistance, high combining ability, high quality, and good out-crossing rateZhao BG, Yuan ZC, Tang L, Huang ZY, Deng XX, Yuan LPChina National Hybrid Rice Research and DevelopmentCenter/Hunan Hybrid Rice Research CenterHybrid rice parent line V20B was transformed with wild rice species Oryza minuta and maize genomic DNA,then backcrossed into YeweiB and 56B. Zi100B with disease resistance and high quality was selected from aYeweiB/FuyiB//56B later generation. By shuttle breeding between Changshas high summer temperature, Sanyalow early spring temperature and Longhuis rice blast disease (Ustilaginoidea Virens) infected fields andsystematic breeding, bing1A was selected from a later generation cross of Zi100B and Ⅱ-32B throughtestcrossing and backcrossing. Bing1A demonstrates a strong combination of advantages: rice blast resistance,good quality, high temperature reproduction, high yielding seed production, ideal plantleaf type, sterile stability,and a good rice genetic background. Genetic and molecular biology research shows that Bing1A has good 176
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京genetic character and meets the requirement of modern seed industries and production. It’s marketingexploration potential is lager. Bing1A has passed Validation of Hunan province.S 5.20 - Stably-expressed Main Effects of the Reciprocal Introgressed Fragments on Grain Weight andShape Traits in rice (Oryza sativa L.)Zheng TQ1,4, Wang Y 1,4, Zhu LH1, Zhai HQ 1,Mei HW2, Xu JL 1, Li ZK 1, 31.Institute of Crop Sciences/National Key Facility for Crop Gene Resources and Genetic Improvement, ChineseAcademy of Agricultural Sciences, 12 South Zhong-Guan-Cun St., Beijing 100081, China;2.Shanghai Agrobiological Gene Center, 2901 Beidi Road, 201106 Shanghai, China;3.International Rice Research Institute, DAPO Box 7777, Metro Manila, Philippines.* Email: xujlcaas@yahoo.com.cn; zhkli@yahoo.com.cn.In this work, reciprocal introgressed fragments stably affecting grain weight and shape traits in three continuousseasons were re-assayed by 142 polymorphic SSR markers belonging to 64 chromosomal bins in a set ofreciprocal introgression lines derived from a cross of Lemont/Teqing of rice (Oryza sativa L.), with 201 lineswith Lemont background (LT-IL) and 252 lines in Teqing background (TQ-IL) in advanced generations (F11~13).Main interesting points were found as following:The remained heterozygote ratio was 5.7 ± 5.9% and 3.1 ± 3.0% in LT-ILs and TQ-ILs, respectively, which wasalmost the same as before in the TQ-ILs (3.18%) but a bit higher than that in the LT-ILs (3.81%) of earliergenerations.A total of 43 (about 67.2%) bins were commonly significantly associated with the stably expressed grain weight(TGW) and grain shape (GS) trait loci in both LT-IL and TQ-IL. Of these commonly detected bins, the stablyexpressed main additive effects were half reverse for TGW between the LT-ILs and TQ-ILs, but was almost allreversed for the GS component traits.Four identified grain shape genes: gw2, GS3, GIF1, and qSW5 were physically near to markers on bins 2.2, 3.4,4.3, and 5.2, respectively. A total of 38 cases for these gene linked markers were found to be significantlyassociated with the grain weight and shape traits in the reciprocal introgression lines. Most of them (92.1%)behaved as additive loci.We used the genetic distance based on crossover rate to roughly assess the length of the chromosomal fragmentscontaining the reported four genes for TGW and GS, the chromosomal fragments containing gw2, GS3 andGIF1 in LT-IL (20.6cM for gw2, 28.5cM for GS3, and 16.2 cM for GIF1) were usually longer than those inTQ-IL (13.7 cM for gw2, 5.3 cM for GS3, and 9.7 cM for GIF1). This could explain why the stably expressedmain effects of these genes were not detected in LT-IL but found in TQ-IL by the probing with the same flakingmarkers. As for qSW5, the lengths of the fragments containing the gene-linked markers are shorter in LT-IL(11.6 cM) than that in TQ-IL (15.7 cM), thus, qSW5 was found to be functioning in both sets of ILs.S 5.21 - Transmission and molecular characteristics of Southern rice black-streaked dwarf virus, a newFijivirus threating rice production in AsiaZhou GH , Wang Q , Pu LL and Zhang SGLaboratory of Plant Virology, South China Agricultural University, Guangzhou, China.* Email: ghzhou@scau.edu.cn 177
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Southern Rice Black Streak Dwarf Virus (SRBSDV, genus Fijivirus, Family Reoviridae), a new virus infectingrice discovered in 2001 in Guangdong province, spread raphidly throughout southern China and northernVietnam, and became one of the most important virus threating rice production in Southeast Asia. The virus wastransmitted by the white back planthopper (WBPH, Sogatella furcifera, Hemiptera: Delphacidae) , along-distance migration rice pest, in a persistent manner. The virus replicates in the WBPH body, but can not betransmitted to offspring through egg. About 60% WBPH individual nymph at 4-5th instar stage was viruliferousin the population developed on the infected rice plant. Both nymph and adult transmit the virus while the formerwith higher effieciency. Acquisition and inoculation peorids were at least 10-30 minutes, and incubation peoridwere 5-7 day when the virus was artificially transmitted through WBPH nymph. The complete genomesequences of this virus, containing ten linear segments of double-stranded RNA, were determined. Based on thenucleotide identity analysis, SRBSDV was most closely related to MRDV and RBSDV. Comparsing to RBSDV,S1, S2 and S10 were most conserved with identities of 78.5-79.2% while S5 and S6 were the least conservedwith 70.6-71.6% identities. No reconbination was found between this virus with related viruses. Transmissionand molecular characteristics revealed in this study provided a base for virus resistance screening and breeding,especially by molecular strategy.S 5.22 - Efficient production of selectable marker free Bt transgenic rice via cis-activatingtransposon-mediated transgene relocationZhou J, Li J, Li YL, Liu ZL and Qu SH *Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Shiqiao Road 198, 310021,P.R.China.* Email: squ111@163.comWe developed an Ac-Ds cis-two-element vector system in which a T-DNA vector contains an immobilized Actransposase gene along with the Ds inverted repeats for transgene relocation. A GFP fluorescence gene used as anegative selection marker against the immobilized Ac and the HPT gene as a positive selectable marker geneconferring hygromycine resistant. The modified hybrid Bacillus thuringiensis (Bt) ð-endotoxin gene Cry1Ab/Acwas used to develop marker-free transgenic Bt rice (Oryza sativa L.) via this high efficient system. Ds-Btexcision frequency was analyzed both in HPT resistant calli and leaves of T0 plants. High excision activity anddiverse excision footprints were discovered in HPT resistant calli, and also in leaves of 29 T0 plants out of total45 independent transformants (64.4%). By PCR screening of the GFP negative T1 plants of 8 transformed T0lines, 3 of them successfully segregated selectable marker-free (SMF) transgenic T1 plants. Investigation of theexpression of Bt transgene in these marker-free lines is undergoing include molecular analysis and bioassay oninsecticidal activity. The diversity and stability of the T-DNA free Ds reposition site will be further analyzed.These preliminary results indicate this cis-activation transposon-mediated marker-free system has the potentialto become a convenient tool for efficient production T-DNA integration site-free transgenic plants.S5.23 Pyramiding QTLs resistance for sheath blight derived from two parents in rice restorer lineShuhui527Gao XQ1, Xie XW1, Xu MR1, Zhu LH1, Zhou YL1*, Li ZK1,2 178
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京1 Institute of Crop Sciences, Chinese Academy of Agricultural Sciences / National Key Facility for Crop GeneResources and Genetic Improvement, Beijing 100081, China; 2International Rice Research Institute, DAPOBox 7777, Metro Manila, Philippines* Email: zhouyl@caas.net.cnSheath blight, caused by Rhizoctonia solani, is one of the most important diseases of rice. Rice sheath blightresistance (SBR) is a complicated quantitative trait and no major resistance gene has been identified in rice,there is little advance in rice SBR breeding. Pyramiding the SBR-QTL derived from different donor thoughmolecular-associated selected is an effective measure to improve the rice SBR.Two near isogenic introgression line in BC2F5 selected from the progenies between shuhui527 as a recurrentparent and two moderate SBR varieties, JCSM and MGDMG as donors, were used for QTL mapping of SBR.Three lines carrying single QTL and agronomic traits similar to Shuhui 527 were selected to be pyramid. Theresults indicated that the pyramiding lines exhibited distinct higher level of resistance for sheath blight than theiroriginal lines with single QTL, which carrying Qsh7c derived from MGDMG and Qsh6b derived from JCSM,Qsh8a derived from MGDMG and Qsh6b, respectively, indicating that pyramiding non-allelic SBR-QTLs fromdifferent resistance donors could improved the level of resistance to sheath blight.. Similarly, pyramiding linecarrying Qsh7c and Qsh8a derived from the same donor decreased 1 disease grade compared with its originallines.S 6 Studies on non-tissue culture plant genetic transformationSun Y, Wang JX, Yang LY, Cui GM, Du JZ, Hao YS and Wang YXAgri-Biotechnology Research Center of Shanxi Province, 030031 China.Email: sunyi692003@yahoo.com.cnGenetic transformation is a powerful tool for plant breeding and genetical, physiological or biochemicalresearch. Two methods are currently widely used for producing transgenic plants, namely Agrobacteriummediation and particle bombardment. However, both of them need tissue culture procedures, which are labor-and time-consuming, tedious and expensive. We have been attempting non-tissue culture or in planta planttransformation methods for more than one decade and successfully obtained transgenic plants with followingapproaches.Pollen-mediated transformation of maize with assistance of ultrasonication The genetic transformation wasachieved by pollen-mediated approach on maize (Zea mays L.) inbred lines, Tai 9101 and Zong 31. PlasmidDNA of pCL II-RC-1 was mixed with fresh pollen of the maize inbreds in sucrose solution. The pollensuspension were treated with ultrasonication, collected, and pollinated on silks of maize ears. Transformantswere confirmed by dot bolt hybridization, PCR amplification and PCR-Southern blot hybridizationAgrobacterium transformation of maize germinating seeds Germinating seeds of maize (Zea mays) werewounded in the meristem area using a scalpel, and co-cultivated with an Agrobacterium tumefaciens strainharbouring a Ti plasmid. Seedlings produced from the treatment were screened by hygromycin selection. Fertiletransgenic T0 and T1 plants were obtained. PCR amplification, PCR–Southern and Southern-blot analysisshowed that the foreign gene had been introduced into the inbreds of maize. About 29% of T0 seedlingsexamined were confirmed to be transgenic, although the overall transformation was only 0.6% when total 179
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京treated seeds were taken into account.In situ transformation of poplar transgenic poplar shoots were obtained using Agrobacterium-mediated in situbud transformation. The method is demonstrated for the first time for poplar. Buds of Populus cathayana Rehdwere co-cultivated with an Agrobacterium strain LBA4404 harbouring the binary vector pBI101-Bmk-chi. Withthe procedure described, transformation efficiencies of 1% and 2.24% were achieved in 2006 and 2007,respectively. Stable integration of the transgene sequence was confirmed by PCR and Southern hybridization.The transgenes could be inherited by vegetative propagation. The novel approach is especially useful for speciesor cultivars not responding to tissue culture. (Scandinavian Journal of Forest Research 25: 3-9).Advantages of in planta transformation are: obviate tedious tissue culture procedures; rapid, thus time-saving;economical; labor-efficient; genotype independent; simple and ready to be integrated into conventional breedingprograms; and easily to be adopted in marker free transformation. Transformation efficiencies of the methodsvary between 1-40%. Studies for further enhancing transformation efficiency are under way.S 7.01 - Application of MAS for resistance to Fusarium head blight in a wheat breeding programCao W, Fedak G, Somers D, Voldeng H, Savard M and Xue AEastern Cereals and Oilseeds Research Centre, AAFC, Central Experimental Farm 960 Carling Ave. Ottawa,ON K1A 0C6. (D.S.) Cereal Research Centre, 195 Dafoe Road, Winnipeg MN R3T 2M9.The objectives of this study were to: 1) determine the effectiveness of marker assisted selection (MAS) relativeto conventional visual selection (CVS) for resistance to FHB in a spring wheat backcross breeding program and2) develop white seeded wheat with a high level of FHB resistance using MAS. BW301, a line susceptible toFHB, was crossed with HC374, a line resistant to FHB and the F1 backcrossed to BW301. A MAS population(MAS BC2F5) was developed through F2-derived method, while two conventional visual selection populations(CVS BC1F6 and BC2F5) have been developed though single seed descent. Seven lines with all three FHB QTLswere selected from the MAS population, while the top 10 resistant lines were selected from CVS BC1 and CVSBC2 populations, respectively, based on FHB symptoms in the FHB nursery of 2005. The 27 lines, plus twoparents, were further evaluated for FHB resistance in a four replicate field experiment in 2006 and 2007. Acombined analysis from two years data showed that the means for FHB incidence, severity and index anddeoxynivalenol (DON) content for the MAS population were 39.2%, 27.3%, 11.2% and 4.6 ppm, for the CVSBC1 population the values were 43.0%, 30.8%, 14.4% and 6.1 ppm and; for the CVS BC2 population the valueswere 50.6%, 41.2%, 21.6% and 9.1 ppm. Marker profiles showed that the lines in the CVS BC1 populationcarried from 0 to 2 QTLs whereas none of the lines in CVS BC2 population carried the FHB resistance QTLs.The results confirmed that MAS is more effective than CVS for improvement of resistance to FHB in this wheatbackcross breeding program and indicated that the more backcrosses, the greater the risk of loss of resistancegenes in backcross breeding if the CVS is used. The results also suggested that minor genes have an importantrole in determining resistance to FHB in wheat.For development of white seeded wheat with resistance to FHB, Snowbird, a FHB susceptible white hard whiteseeded wheat, was crossed to Sumai 3 as a female parent. Twenty thousand F2 plants were produced and grownin the greenhouse. One thousand and five hundred white seeds were visually selected from the F2 population.This population was advanced to F5 by single seed descent. At the seedling stage of F5, a MAS was performed 180
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京for three FHB QTLs on chromosome 5A, 3B and 6B. Two hundred and fifty F5 lines were selected with two orthree resistance QTLs and grown in a FHB nursery in 2008. Fifteen F6 lines were selected based on FHBresistance and agronomic performance. Seed of the 15 lines was increased in the greenhouse in the winter of2009. These 15 lines and two parents Sumai 3 and Snowbird, plus AC Vista as a check were planted in the FHBnursery with three replications and in a preliminary yield trial with two reps in the summer of 2009. The resultsshowed that several white seeded wheat lines had high levels of resistance to FHB; significantly higher thanSnowbird and matured earlier than Sumai 3. Quality is also improved significantly compared to Sumai 3, basedon the Glutomatic test.S 7.02 - Quality improvement for wheat cultivar Lunxuan 987 using molecular markersChen XM, Xia XC, Zhang Y, He ZH, Zhang Y, Wang DSInstitute of Crop Science, National Wheat Improvement Center, Chinese Academy of Agricultural Sciences(CAAS), 12 Zhongguancun South Street, Beijing 100081, China.E-mail: Chenxm@mail.caas.net.cnYield increase and quality improvement are the most important breeding objectives. Generally, it is very hard toimprove both yield and quality simultaneously because there is a negative correlation between them. However,it may be possible to obtain this goal by molecular marker assisted breeding. Although some studies pointed outthe limitation of high molecular weight glutenin subunits (HMW-GS) in predicting the end-use quality, theHMW-GS play a key role in determining bread-making quality, particularly some HMW-GS, such as 5+10, aremore important than others. The Yumai 34, a high bread-making quality and low yield wheat cultivar withHMW-GS 1, 7+8, 5+10, was released in Yellow and Huai River Winter Wheat Region. Lunxuan 987 has highyield, but poor bread-making quality with HMW-GS 1, 20, 2+12, and it is released in North Winter WheatRegion. In the present study, Yumai 34 as a donor for good quality and Lunxuan 987 as a backcross parent wereused to make a cross and molecular markers for HMW-GS 7+8 and 5+10 were used in backcross selection. FourF5 lines with both 7+8 and 5+10 were developed from the cross Yumai 34/Lunxuan 987*3. The quality of thefour lines is improved significantly, with grain hardness from 71.1 to 77.3, grain protein content from 13.0% to13.9%, mixing time from 3.9 min to 4.6 min, right slope from -0.5 to -2.1, timer width at 8 minute from 10.6mm to 13.8 mm, whereas all the corresponding quality indices for Lunxuan 987 are 65.1, 13.7%, 1.6 min, -1.4and 3.2 mm, respectively. Meanwhile, all the quality indices for the good quality control Zhongyou 9507 are 65,15.0%, 6.2 min, -1.5 and 14.5 mm, respectively. The yield test in year 2010 showed that the yield of the fourlines were from 5885 kg/ha to 7233 kg/ha. The lines 40555 and 50564 produced 7233 kg and 7106 kg perhectare, which are 14.7% and 11.3% higher than the check cultivar Jingdong 8, respectively. These twopromising lines showed high yield, good quality, lodging resistance, powdery mildew resistance, and goodlooking at maturity. This study indicated that yield and quality could be improved at the same time in molecularmarker assisted breeding.S 7.03 - Comparative Genetic Mapping of Powdery Mildew Resistance Locus Pm5 in Common Wheat(Triticum aestivum L.)Li JT, Zhang HT, Zhang D, Sun QX, Liu ZYState Key Laboratory for Agrobiotechnology, Dept.of Plant Genetics & Breeding, China Agricultural University, 181
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Beijing 100193, P.R.ChinaPowder mildew, caused by Blumeria graminis f. sp. tritici, is one of the most important diseases in wheat.Breeding and growing resistant cultivars are the most economical and effective methods to reduce crop yieldlosses due to this disease. Five alleles for Powder mildew resistance locus Pm5 (Pm5a-Pm5e) have been locatedon 7BL chromosome. Powder mildew resistance loci found in many Chinese landraces or cultivars, i.e.Xiaobaidong (xbd), Hongquanmang (PmH), Fuzhuang 30 (Pm5e), Tangmai 4 (PmTm4) etc are supposed to beallelic or closely linked to Pm5. In order to develop high density linkage map of Pm5, we performedcomparative genomic analysis between the Pm5 genetic region and rice and brachypodium genome sequences.The results revealed that the genomic regions flanking Pm5e and PmTm4 in wheat showed high co-linearitywith the distal of rice chromosome 6L and the brachypodium chromosome 1. Wheat ESTs orthology toBrachypodium orthologous genomic region were applied to develop polymorphic EST-STS markers to mapPm5e using F2:8 recombinant inbred lines of Fuzhang30/ Nongda015 and to map PmTm4 using F2:3 families ofTangmai 4/ Clement .Two and five polymorphic EST-STS makers were founded to be linked to Pm5e and PmTm4 respectively, Pm5ewas closely linked to BD410 at a genetic distance of 1.5 cM, and PmTm4 was linked to TC80 at a geneticdistance of 7.6 cM.S 7.04 - Mapping QTLs for Seedling Heat Tolerance in Common wheatLi SP 1,2,3, Jing RL1, Wang CS2, Chang XP 11 Institute of Crop Science,Chinese Academy of Agricultural Sciences/Key Laboratory of Crop Germplasmand Biotechenology, Ministry of Agriculture, Beijing 100081;2 College of Agronomy, Northwest A &F University, Yangling Shaanxi 712100;3 Wheat Research Institute, Shanxi Academy of Agricultural Sciences,Linfen Shanxi 041000.E-mail: jingrl@caas.net.cnA doubled haploid(DH) population with 150 lines, derived from a cross between two Chinese common wheatvarieties Hanxuan10 and Lumai14, was used as the plant material to mapping quantitative trait loci(QTL) forseedling heat tolerance. Several seedling traits including root dry weight( RDW),shoot dry weight (SDW),ratioof RDW/ SDW, Chlorophyll fluorescence parameters(CFP)(Fv/Fm) and chlorophyll content(CC) weremeasured under heat stress and normal condition respectively. QTLs and G×E interactions for these traits weredetected using mixed-model-based composite interval mapping method. A total of five additive QTLs and 12pairs of epistatic QTLs associated with the target traits were mapped on chromosome 1D, 2A, 2B, 2D, 3A, 3B,3D, 4A and 6B,respectively. Among them, two QTLs with additive effect were identified controlling shoot dryweight, three for seedling biomass,and one pair of epistatic QTL for root dry weight, five pairs for shoot dryweight,three pairs controlling seedling biomass, two pairs for chlorophyll content and one pair for Chlorophyllfluorescence parameters,respectively. Four additive QTL×environment and six pairs of epistaticQTL×environment interaction effects were also identified. The results may be helpful for marker-assistedselecting seedling traits related to heat tolerance in wheat. 182
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京S 7.05 - Creating and identification of wheat-alien substitution lines involving homoeologous group 2chromosomes from related genus by EST-PCR markersLin ZS, Ye XG, Niu YZ, Shen ZW, Wang MJIn order to develop substitution lines involving in alien chromosome from Thinopyrum intermedium andHordeum, six crosses containing 2Ai-2, 2H were made. The crosses include three types: (1) wheat-Thinopyromintermedium substitution lines 2Ai-2(2D) N431, N452, respectively, crossing with wheat-barley substitution line2H (2B); (2) wheat-barley substitution line 2H (2D) crossing with wheat-Th.intermedium substitution lines2Ai-2 (2B) N420, N439, respectively and (3) wheat-barley substitution line 2H (2A) crossing withwheat-Th.intermedium substitution lines 2Ai-2 (2B) N420, N439, respectively. 4 EST-PCR markers were usedto test the hybrids and F2 populations. The plants in F2 populations were approximately divided into 4 genotypesby the existing of the alien chromosomes: ⅰ) no alien chromosome, ⅱ) only 2H chromosome, ⅲ) only 2Ai-2chromosome and ⅳ) both 2H and 2Ai-2. The frequency of the 4 genotypes in F2 was different largelydepending on the type of the cross, though the chromosome constitute in the F1s of crosses type (1) and (2) wasthe same. The result also showed that the 2B chromosome was easier to be replaced by the alien chromosomecomparing with 2D or 2A in genotype ⅳ). With both alien chromosomes present, 22, 7 and 3 plants absenting2B, 2D, 2A respectively were obtained. Selection of stable substitution lines having 2H and 2Ai-2 chromosomesis underway. The target genetic stocks will be helpful for the research on plant polyploidization.S 7.06 - Characterization and molecular mapping of a stripe rust resistance gene in CIMMYT syntheticwheat Cid 25Ren Q 1, Liu HJ 1, Zhang ZY 1* , Feng J 2 , Pu ZJ 3 , Xu SC 2 , Xia XC 1 and Xin ZY 11 The National Key Facility for Crop Gene Resources and Genetic Improvement, Institute of Crop Science,Chinese Academy of Agricultural Sciences (CAAS), Beijing 100081, China;2 Institute of Plant Protection, CAAS, Beijing 100094, China; 3Crop Research Institute, Sichun Academy ofAgricultural Sciences, Chengdu 610066, China.Email: zhangzy@mail.caas.net.cnStripe rust, caused by Puccinia striiformis f. sp. tritici (PST), is one of the most destructive diseases of wheat(Triticum aestivum L.). The best strategy to control stripe rust is to breed and grow resistant cultivars. ACIMMYT synthetic wheat line Cid 25 was identified to be resistant to 30 PST isolates. Seedlings of the parents,F1 plants, F2 plants, and F3 lines derived from the cross Cid 25×Mingxian 169 were tested with race CYR32.Resistance to stripe rust in Cid 25 was controlled by a single dominant gene temporarily designated YrC25. Bysimple sequence repeat (SSR) and bulked segregant (BSA) analyses, a molecular map, harboring YrC25 and 9linked SSR markers, was constructed. YrC25 was flanked by markers Xcfd65, Xbarc187, Xgwm18, Xgwm11,Xbarc137 and Xbarc240, within genetic distances of less than 2 cM. These linked markers and YrC25 werelocated on the short arm of chromosome 1B using the Chinese Spring nullisomic-tetrasomic and ditelosomicstocks. The chromosome location, reaction patterns to 30 PST isolates and a test of allelism indicated thatYrC25 was likely to be of the same as Yr24/Yr26. After completion of the research, a new mutant race wasisolated from Chuanmai 42 in Sichuan province. This race is highly virulent for seedlings of Chuanmai 42,virulent for Cid 25, Yr24/3*AvS and Yr26/3*AvS in controlled greenhouse tests, confirming the likelihood of a 183
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京common resistance gene.S 7.07 - The strategy and potential utilization of temperate germplasm for tropical germplasmimprovement:a case study in maize (Zea mays. L)Wen WW1, Guo TT 2, Chavez Tovar Victor H 1, Yan JB 1, 2, Taba Suketoshi 11 International Maize and Wheat Improvement Center (CIMMYT), Apartado Postal 6-640, 06600 Mexico, DF,Mexico; 2 National Maize Improvement Center of China, China Agricultural University, Beijing 100193, ChinaOrganization of maize germplasm into genetically divergent heterotic groups is needed for breeding highyielding hybrids by exploitation of heterosis. In this study, 94 CIMMYT inbred lines (CML) and 54 USGermplasm Enhancement of Maize (GEM) lines were assembled and characterized using 1,266 SNPs with highquality. Based on principle component analysis (PCA), GEM and CML lines were clearly separated. There weretwo groups of GEM lines classified by PCA that seemed to correspond to the stiff stalk (SS) and non-stiff stalk(NSS) heterotic groups bred by GEM. CML lines did not form obvious subgroups by PCA. Each of 1,266 SNPallele frequency differed in GEM and CML. 3.6% alleles (46/1,266) of CML were absent in GEM, and 4.4%alleles (56/1,266) of GEM were absent in CML. CML heterotic group A with GEM heterotic group SS and CMLheterotic group B with GEM heterotic group NSS were crossed and developed 654 F1. Association mappingwas performed based on both 148 CML, GEM lines and 654 F1s for seven agronomic traits at two locations.For plant height (PH) and anthesis time (AT), genomic estimated breeding values (GEBVs) for a testing set of215 F1s were predicted based on the training data of 430 F1s using a best linear unbiased prediction (BLUP)method. The accuracy benefitted from the adoption of markers associated with QTLs for both traits. However, itdoes not necessarily increase with the raise of associated marker density. High correlations between actual andpredicted phenotypic values of F1 indicated the possibility to choose high performing parental combinations ofphenotypic traits using SNP markers. It is hoped that we can enhance the tropical maize germplasm byincorporating temperate germplasm more efficiently, through genome wide selection. Clear heterotic patterns ofthe GEM lines and unique alleles can facilitate tropical maize hybrid breeding.S 7.08 - Quantitative trait loci for carbon isotope discrimination in wheat DH populationWu XS, Jing RL* and Chang XPNational Key Facility for Crop Gene Resources and Genetic Improvement/Key Laboratory of Crop Germplasmand Biotechnology, Ministry of Agriculture/Institute of Crop Science, Chinese Academy of AgriculturalSciences, Beijing 100081, China.* Email: jingrl@caas.net.cnCarbon isotope discrimination (∆) has been suggested as a useful indicator for indirect selection of grain yieldand water use efficient in cereals. To enhance the selection efficiency of ∆, it is of great importance to evaluatethe genetic variation for ∆ and the relationship between ∆ and grain yield. A double haploid (DH) populationderived from Hanxuan 10 and Lumai 14 were phenotyped for ∆ in flag leaf, grain yield, yield components andplant height (PH) in two trials contrasted for their water availability, rain-fed and irrigated environments inChangping, Beijing in 2007. Quantitative trait loci (QTLs) were identified by composite-interval mapping(CIM). Five putative QTLs for ∆ were detected on chromosomes 1A, 5A, 7A and 7B, and four to nine QTLs for 184
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京other related traits were founded on all chromosomes except for 2B, 5B, 5D and 6D. No consistent QTLsbetween two water regimes for all traits except TGW and PH indicated that their QTL expressions were easilyaffected by environments. QTL clusters for grain yield with number of grains per spike (NGS) and PH, grainyield per plant (YP) with number of spikes per plant (NSP), NGS with one-thousand grain weight (TGW), andNSP with PH, possibly owing to pleiotropism and linkage, could explain the correlations between different traits.In the vicinity (10 cM ~ 20 cM) of QTL for ∆ on 5A (near WMC74) and 7A (near WMC488), there wererespectively a QTL controlling YP and NSP, their same contributer Hanxuan 10 suggested their genetic linkage.Compared to the low ∆ progeny, a group of progeny with high ∆ had an increased frequency of few QTL forhigh ∆, grain yield and NSP under the rainfed condition and only for high ∆ and NSP under the irrigatedcondition, which was consistent with agronomic exhibition. Thus, selection for ∆ appears beneficial inincreasing grain yield in rain-fed environments in our study.S7.09 FISH analysis of a CRW –homology sequence from Pseudoroegneria spicata in Thinopyrumponticum and Th. intermediumLu K1,2, Xu Z2, Liu Z1, Zhang XY11 Key Laboratory of CropGermplasm&Utilization,InstituteofCrop Sciences, Chinese Academy of AgriculturalSciences, Beijing 100081,China; 2 Key Laboratory of Grassland Resource and Ecology, Ministry of Agriculture,Grassland Research Institute, Chinese Academy of Agricultural Sciences, Huhhot 010010, China.To verify the genome components of Thinopyrum ponticum Liu & Wang and Th. Intermedium [Host] Barkworth& Dewey,six specific primer pairs were designed according to the sequence of an autonomous centromericretrotransposon of wheat(CRW) from Triticum boeoticum. Several DNA fragments were amplified by PCR fromthe genomic DNA of the diploid species Pseudoroegneria spicata A Love. After sequence alignment, a 1.755 kbfragment was obtained and named as pStCl (St Genome centromere associated sequence, GenBank accessionNo.FJ952565). This fragment contained an 800 bp fragment highly homologous to the LTR region ofautonomous CRW, a short fragment partial homology to the gag region of CRW, and an AGCAAC—richtandem repeat.Fluorescent in situ hybridization (FISH) using pStC 1 as a probe was carried out on thechromosomes of Th. ponticum, Th. intermedium and Triticum aestivum CV.Chinese Spring. Th.ponticum wasproved to have two St and three E genomes(St1St2EeEbEx). In Th. Intermedium, strong FISH signals wereobserved on St genome chromosomes, while faint signals were also found on some E genome chromosomes attheir pericentromeric regions . These results indicated that during speciation of sub-genomes in theallopolyploids of Thinopyrum genus,concerted evolution might have occued at centromeric and pericentromericregions.S 8.01 - Identification, transferability and application of novel EST-SSRs in Hevea brasiliensisAn ZWa,b, Cheng Ha, Zhao Y H c, Hu YSa, L iWGa, Huang HSa, *a State Center for Rubber Breeding, Rubber Research Institute, Chinese Academy of Tropical AgriculturalSciences, Danzhou, Hainan 571737, China.b College of Agriculture, Hainan University, Haikou, Hainan 570228, China.c College of Life sciences, Ludong University, Yantai, Shandong 264025, China.* Email: xjshhs@163.com 185
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Expressed sequences tags derived simple sequence repeats (EST-SSRs) are rarely used in rubber tree, but havebeen used for a variety of purposes in other plants. 18,218 unigenes were obtained from 26,838 Heveabrasiliensis ESTs containing 16,838 cold-stressed ESTs and 10,000 ethylene-stimulated ESTs. 2,370 SSR locidistributed in 1,888 unigenes which accouted for 10.4% of 18,218 unigenes. The average distance of one SSRwas 4.08 kb of EST sequences. Mononucleotide, dinucleotide and trinucleotide repeats were the dominant typesamong the identified SSRs, accounting for 55.4%, 23.6% and 12.1%, respectively. A/T, AG/CT, and AAG/CTTwere the most abundant motifs for mono-, di- and trinucleotide motifs. 500 primer pairs were designed byPRIMER5.0 and 32 primer pairs were synthesized. Among which, 22 primer pairs were successfully amplifiedin 69 wild accessions of H.brasiliensis and four relative species, H. pauciflora, H.nitida, H.spruceana,H.camargoana. All of the 22 primer pairs showed perfect transferability and high polymorphism. The geneticstructure of H.brasiliensis wild populations were investigated by 22 EST-SSRs. The results showed that thedifference of each parameter wasn’t significant among population Acre(AC), Rondonia(RO) and MatoGrosso(MT), and population MT had more diversity than the others to some extent. The genetic differentiationof H.brasiliensis wild populations (GST) was 0.18, and gene flow (Nm) was 1.43, which indicated that thegenetic difference of H.brasiliensis wild populations mainly came from the individuals rather thaninter-populations. The study suggested that EST-SSRs are superior to genomic-SSRs, and can be successfullyused for a variety purposes in rubber breeding program.S 8.02 - Polymorphic Identification of PCR Based Markers Linked to Heading Date Controlling Genes toAid Selection in Developing Ultra Early Maturing VarietyEnung Sri Mulyaningsih, Untung Susanto, Bambang SuprihatnoVery early maturing variety is an important tool to avoid drought stress. It could be panted just after first wetseason planting and could be harvested at the early f dry season without suffering any drought stress yet unlikethe relatively longer growth duration rice variety. This approach could increase crop index and in turn increaseproduction and farmer’s income. Heading date is a major trait affecting rice growth duration, so that it couldbe used as a main approach to breed ultra early maturing rice variety (less than 90 days). This research wasaimed to assess variability on heading date related alleles among various growth duration rice varieties as a basestudy to pyramid those genes. For that purpose, forty three published PCR based markers linked to headingdate genes has been applied toeight varieties consisted of six very early maturing variety (less than 104 days ofgrowth duration; Silugonggo, Inpari 13, Slegreng, Srijaya, Mudgo, and Milky Rice), one early maturing variety(Ciherang), and one medium maturing variety (less that 125 days of growth duration; Cisadane). The resultsshowed that only 7 (16,3%) of the markers showing polymorphism. Development of dendrogram usingEuclidian distance divided the varieties into three groups. Ciherang (very early maturing variety) andCisadane (medium maturing variety) grouped together as group 1, and the rest very early maturing varietieswere divided into two groups. Milky rice and Silugonggo grouped together as group 2 and the rest fourvarieties (Srijaya, Slegreng, Inpari 13, and Mudgo) as group 3. It showed that different growth duration hasdifference allele of heading date controlling genes. On the other hand, there is variation among short growthduration variety. This variation could be used as guide to pyramid the gene to develop ultra early maturingvariety. These results indicated that cross combination between group 2 and 3 would combine the heading date 186
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京controlling genes. The identified polymorphic markers could be used to aid the selection to pyramid the genes.S 8.03 - TP-M13-SSR technique and its applications in the studies on the analysis of genetic diversity forpear germplasm resourcesGao Y, Tian LM, Liu FZ, Cao YF*Research Institute of Pomology, CAAS/Key Laboratory of the Use of Fruit Germplasm Resources, Ministry ofAgriculture, Xingcheng 125100, ChinaEmail: yfcaas@263.netAn economical method of simple sequence repeat (SSR) detection was used in studies on genetic diversityanalysis of 25 cultivars of 5 species in which fluorescent labeling of SSR-PCR products was used, with tailedprimer M13 (TP-M13-SSR). Two pairs of primers NH013a and NH007b could distinguish all of the pear cultivars,the identified rate of 5 pairs of primers is 76% on average.The ranges of gene diversity, PIC and locusheterozygosity on 5 SSR locuses were 0.6560~0.83460, 0.6398~0.8149 and 0.4348~1.0000 respectively. Thegene diversity and PIC of P. sinkiangensis on average which were 0.7160 and 0.6812 respectively were the lowest,but those of P.Ussuriensis which were 0.7720 and 0.7379 respectively were the highest.The method has theadvantages of high-throughput, sensitiveness, cost-effectiveness and high accuracy. And it had been used instudies on genetic diversity analysis of pear successfully. Therefore, the TP-M13-SSR automated fluorescentdetection system is suggested to be suitable for studies on genetic diversity analysis of many pear accessions bythe use of a large number of SSR markers. The same as other accessions with large genome size, the TP-M13automated fluorescent detection system can also be attempted.S 8.04 - Performance prediction of hybrids between recombination inbred lines in maizeGuo TT *§, Li HH§, Tang JH¶, Li JS*, Zhang ZW‡, Bernardo R†, Yan JB *‡, Wang JK§‡*National Maize Improvement Center, China Agricultural University, China. §Institute of Crop Science,Chinese Academy of Agriculture Sciences, China. †Department of Agronomy and Plant Genetics, University ofMinnesota, USA. ¶Henan Agricultural University, China. ‡International Maize and Wheat Improvement Center(CIMMYT), Mexico.* Email: Wangjk@caas.net.cnAlong with the development of larger breeding programs have come motivations for breeders to create moreefficient methods of identifying potential hybrids among numerous parental combinations. Here we attempted toconstruct statistical models using information including molecular markers and phenotypic records of 11 traitsfrom 294 recombinant inbred lines (RILs) and 441 immortalized F2 hybrids (IF2). Four models wererespectively based on inbred line per se performance (ILP), general combining ability (GCA), QTL mapping(QTL) and genome-wide selection (GWS). Inclusive composite interval mapping (ICIM) was implemented toestimate QTLs effect for QTL method and best linear unbiased prediction (BLUP) could be utilized to estimateeffect of markers on the whole genome for GWS method. In data analysis, 294 RILs and 2/3 hybrids selectedrandomly were assigned into training set; the remaining 147 hybrids were in testing set. Results of validationshowed: the correlations between actual and predicted phenotypic values were 0.359-0.844 for ILP, 0.486-0.776for GCA, 0.231-0.602 for QTL, 0.579-0.804 for GWS, from which GWS had high potential to estimate 187
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京genotypic values of yet observed hybrids accurately. However, when breeding values were estimated as additiveeffect using kinship and marker matrices in the first step, and dominant effect subsequently using only markers,results demonstrated diversely among kinds of methods: 0.521-0.819 for ILP, 0.559-0.868 for GCA,0.173-0.782 for QTL, 0.541-0.823 for GWS. There was no significant difference between methods of GCA andGWS. Taking grain yield as an example, correlation was 0.606 for GCA, and 0.660 for GWS. Theoretically,GCA of inbred lines consisted of additive effect, partial dominant effect as well as epistasis. Our results suggestthat prediction based on GCA is also efficient in practical breeding on the condition that estimation of breedingvalue is credible. Furthermore, for Grain yield, there were 114 combinations of which performances were betterthan original F1 with high heterosis. It was implied that it’s possible to select superior combinations prior toevaluate them in field trials. Thus, the prediction of parental combinations will facilitate breeding process byavoiding the time-consuming and labor-intensive trait evaluation.S 8.05 - Potato starches with altered properties and morphologyHuang XF 1,2, Nazarian FF1,3, Jean-Paul V1,4, Qin J1,5, Suurs LCJM1, Visser RGF1 and Trindade LM11 Wageningen UR - Plant Breeding, Wageningen University and Research Center, P.O. Box 386. 6700 AJWageningen. The Netherlands.2 Graduate School Experimental Plant Sciences3 Present address: Agronomy and plant breeding group, Faculty of Agriculture, University of Lorestan, P.O.Box465, Khorramabad, Iran4 Present address: Laboratory of Food Chemistry, Wageningen University, P.O. Box 8129. 6700 EVWageningen. The Netherlands.5 Present address: Department of Biology, Huaiyin Teachers College, 223300, Huaian, ChinaStarch is the most important and abundant storage carbohydrate in plants. It is mainly composed of twoα-glucan polymers, linear amylose (20-30%) and highly branched amylopectin (70-80%). Starches and starchderived products are used in food, feed and numerous industrial applications. Before starches can be used formost industrial applications, they need to be modified in order to overcome physico-chemical limitations.Modification of starch biosynthesis pathways holds an enormous potential for tailoring granules or polymerswith new functionalities. Until recently a lot of effort was put into investigating the individual components inthe starch biosynthetic pathway. One successful strategy used in order to alter the starch granule structure wasthe tagging of different enzymes or enzyme domains to the starch granule using microbial starch-bindingdomains (SBDs).Our lab has demonstrated that microbial starch-binding domain (SBD) successfully accumulated in starchgranules during starch granule biosynthesis, without affecting starch physico-chemical properties (Ji et al.,2003). Using SBD as an anchor tool different microbial enzymes, which have desired function onoligosaccharides and/or starch, were introduced in potato to produce starches with novel structures andproperties.An SBD of cyclodextrin glycosyltransferase from Bacillus circulans was fused with two different bacterialenzymes, the amylosucrase (AS) from Neisseria polysaccharea and the E.coli glycogen branching enzyme(glgB). Expression of SBD-amylosucrase fusion protein in the amylose containing potato resulted in starchgranules with a rough surface with increased granule size, changed physico-chemical properties, and higher 188
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京enzymatic digestibility. The presence of GLGB-SBD fusion protein resulted in both amalgamated starchgranules and punctured starch granules with relatively large holes in the amylose-free (amf) genetic background.In the amylose-containing background, starch granules showed an irregular rough surface.By altering the starch composition and structure, granular starches can be obtained with novel physicalproperties and a broad range of industrial applications.S 8.06 - Application of Marker-Assisted selection for Striga resistance in cowpea (Vigna unguiculata L.WALP)Jean Baptiste S. Tignegre, Muranaka Satoru, Jeremy T. Ouedraogo, Issa Drabo, Ousmane Boukar, N Moutari,Mamadou Toure.INERA Burkina Faso, IITA, INRAN Niger, IER MaliStriga gesnerioides is a noxious root parasite that can cause complete yield loss if only susceptible varieties areinvolved. The research being initiated in that project aims at applying available AFLP-SCAR markers forselecting Striga gesnerioides-resistant sources in cowpea. To achieve this, the most prevailing two Striga racesin Burkina Faso (race 1), Niger and Nigeria (race 3) were targeted as damaging constraints to address.Participatory variety selection and rural appraisal, including farmers, other end-users, researchers, anddevelopers were conducted. Adapted cowpea resistant lines were implemented in Burkina Faso and Nigeria in2007 and 2008. Farmer’s preferred cowpea cultivars were selected through field-screening and then involved ina backcross selection associated with MAS to combine both Striga resistance and farmer’s desirable traits in asingle variety. An attempt to validate sets of AFLP and SSR markers shows some polymorphisms specific to thecrosses. No marker showed polymorphisms to all combinations of crosses. Currently, in Burkina Faso, Nigerand Nigeria, some lines, though resistant in infested-field, showed bands similar to susceptible cowpea lines. Inshort, back cross populations varying from BC2F1 to BC4F generations and F4 pedigree breeding populationsare being screened during this on-going rainy season.S 8.07 - Analyses of the genetic relationships in 22 species of Manglietia Plant using ISSR markersLi X, Li XL, Ma TY, Wang YB, Chen FYBiotechnology Research Center, China Three Gorges University, Yichang, Hubei 443002, PR China;* Email: chenfj616@163.comThe Manglietia plant is an original group in the existing Magnoliaceae plants, and there are about thirty speciesall over the world, which mainly distributed in tropical and subtropical zones of Asia. There are almost twentyspecies in china, which originated in the southern of the Yangtze River and are the major tree species inbroad-leaved evergreen forests. Since the establishment of this genus, scholars all over the world havecontributed a lot on morphology. Though the poineer work has been done in classifying according to themorphological characters of each species in Manglietia plants, the classification of the inner system inManglietia genus as well as the genetic relationships of different groups or different species have stilldisagreement. The ISSR (inter-simple sequence repeat) molecular-marked technique has generally been used toanalyze the genetic diversity of plants, genetic relationship, origin and evolution, map creation and comparativegenomics. 189
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京The present study applied the ISSR molecular-marked technique to the genetic analysis of 22 species ofManglietia plant, in order to offer molecular evidence for the study of the category of Manglietia and to revealthe genetic relationship of various groups and inter-species in genus at molecular level. The geneticrelationships among 22 species of Manglietia plant were analyzed by using ISSR molecular-marked technique.Ten ISSR primers were selected to assess the genomes of 22 species of Manglietia plant. The results showedthat a total of 595 DNA bands were amplified and 562 of which (94.45%) were polymorphic. According toNei-Li genetic similarity, UPGMA cluster analysis indicated that the 22 species were classified into 6 clustergroups with the genetic similarity of 0.68, and they were classified into 12 sub-cluster groups with the geneticsimilarity of 0.70. The Nei-Li genetic similarities of 22 species of Manglietia ranged from 0.605(between M.Yuyuanensis and M .conifera) ~0.956 (between M. hookeri and M. wangii).The average genetic similarity is0.778, which suggests that Manglietia have a plenty of inter-specie diversity. The genetic similarity coefficientbetween M. hookeri and M. wangii is 0.956, and their genetic relationship is actually most close. The resultsupports the classification of M. patungensis, M. Yuyuanensis and M. forrestii as independent speciesrespectively.S 8.08 - Screening and identification of the AFLP makers linked to a new powdery mildew resistance genein wheat 6V/6ALi XXP Liu XY, Li WL, Wang ZY*College of Life Science, Tianjin Key Laboratory of Cyto-Genetical and Molecular Regulation, Tianjin NormalUniversity, Tianjin 300387, P. R. Dr. Zhenying Wang, College of Life Science, Tianjin Key Laboratory ofCyto-Genetical and Molecular Regulation, Tianjin Normal University, Tianjin 300387, P. R. China.* Email: wzycell@yahoo.com.cnPowdery mildew is one of the most serious diseases of common wheat in the world, which dramatically reducethe wheat production and quality. Owing to the single resistance gene source and its repeated use, severalpowdery mildew resistance genes have been lose effect. Therefore,to transfer and utilize the high quality genesthen culture new resistant wheat strains are crucial for Powdery mildew prevention. So screening andidentification of the AFLP makers linked to a new powdery mildew resistance gene in wheat is one of the moststraightforward approaches to breeding new resistant wheat strains. Wheat H-.villosa chromosome substitutionline(6V/6A)was bred through hybrid ization of H.villo-sa(VV)with T.durum(AABB), then hybrid F1 waspollinated freely by cultivar wheat(AABBDD). Its resistant near-isogenic lines (NILs) was obtained by donorparent 6V/6A and recurrent parent Jing411. It was thought that the genetic background of NILs and 6V/6A maybe resistant to powdery mildew, and the aim of this work was for screening and identification of the AFLPmakers linked to a new powdery mildew resistance gene in wheat 6A/6V, which will be useful formarker-assisted selection and gene pyramiding in wheat resistance breeding. NILs, 6V/6A and Jing411 wereanalyzed using 120 AFLP primer combinations by Li-COR 4300 analysis system. Between 120 and 180 bandsper strain were obtained, ranging from 40 to 450bp. Selective amplification created 1350 unambiguous bands,of which 14 bands were polymorphic in 8 AFLP primer combinations. Few of polymorphisms detected betweenJing411 and NILs in a large proportion of primer combinations, more polymorphisms obtained in 6V/6A, theseresults suggest that NILs and Jing411 had nearly the same genetic background. Four of them only existed in6V/6A, and five of them obtained in Jing411 and NILs, but not in 6V/6A, these bands may relate to some other 190
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京different characters between Jing411/NILs and 6V/6A but no Powdery mildew resistance. Four bands weredetected only in Jing411, and one of the bands appeared in 6V/6A and NILs, but not in Jing411, the 5 bands maybe interrelated with Powdery mildew. However, further studied need to be done with more primer combinationsand sequence analysis.S 8.09 -The progress of Ramie induced mutation breeding technologyLiao WW, Jie YC *, Du XH and Mei KInstitute of Ramie, Hunan Agriculture University, Chang sha 410128Ramie (Boehmeria nivea (L.) Gaud) (Urticaceae, Boehmeria) is perennial herb, originated in China, for thepurpose of harvesting trophosome, it is used as both fiber and fodder, so we improve characteristics of ramievarieties through ploidy breeding to obtain ramie new varieties of high quality. This paper selectively analyzesthe main methods and achievement about the ramie radiation mutagenesis with 60Co-r rays and chemicalmutagenesis with colchicines. When using radiation mutagenesis, the parents should be selected first, they musthave fine genotype, good agricultural characters and only several characters need to be improved. Then inducemutation by radiation treatment. The mutant will be tested at last. The key of this technology is the selection ofparents, radiation source, radiation dose and position. Chemical mutagenesis uses the method of colchicinescombining with tissue culture, and then induces test tube plantlet. In the research of polyploidy, tissue culturecan improve the inducing frequency of colchicines, survival rate and reproduction coefficient, on the other hand,it can make all kinds of cells survive without being obsolete. Using the mutation breeding technology it hasmade some achievements, “YuanYeQing " and "7469" was the radiation offspring of "XiangZhu 1"with 60Co-rray. In addition, the researchers also use 60Co -r ray to irradiate the tube seedlings and have produced four goodmutant in two years; Since the 1982 ,our institute have used different colchicine to deal with the seed,subterranean stem and tube seedlings of ramie, the polyploid of ramie have been successfully inducted inApril ,1983. The Tri-1、 Tri-2 were the ramie polyploid varieties that have been breeded by the institute of ramie,hunan agriculture university for 20 years. In practice, it has showed that mutation breeding is an importantmeans as a kind of ramie germplasm innovation; it is also quite effective in improving the quality and yield offiber. It is proposed the development direction of ramie induced mutation breeding in our country: radiationmutagenesis combined with chemical mutagenesis, radiation breeding combined with conventional breedingand biological technology, further improving the accuracy of mutation breeding, and strengthening the researchof space mutation breeding technology.S 8.10 - Integrated approach for reliable sample archiving and DNA extraction of plantsQiu JS, McCullough A, and Oommen AGeneSeek, a Neogen Company, 4665 Innovation Drive, Suite 120, Lincoln, NE 68521, USA.* Email: jqiu@neogen.comReliable sample archiving and low-cost DNA extraction are critical for effectively carrying out practicalmarker-assisted plant breeding programs. We have developed a simple approach to archiving leaf samples usinga collection device and a simple, high-throughput DNA extraction procedure. Leaves are collected and placedin a small (5 x 5 cm2) collection device with printed barcodes. Two to three small (~2 mm diameter) dry leaf 191
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京punches per sample were taken directly from the collection device, and the DNA extracted utilizing a modifiedalkaline lysis method. The DNA samples are suitable for routine PCR applications and SNP analyses usingMassARRAY® genotyping system. Our studies demonstrate that the archived leaf samples can be stored atambient temperature and DNA extracted from a wide variety of plant species give robust and reproducibleresults. This simple approach does not require expensive reagents or equipment and is applicable forhigh-throughput genotyping in a variety of plant species.S 8.11 - TRAP markers for assessing genetic diversity of 75 most important sugarcane cultivars inMainland ChinaQue YX , Song XX , Chen RK and Xu LP *Key Laboratory of Sugarcane Genetic Improvement, Ministry of Agriculture, Fujian Agriculture and ForestryUniversity, Fuzhou 350002, China.* Email: xlpmail@yahoo.com.cnMarker-assisted selection is one of the most crucial approaches in sugarcane improvement program. Geneticdiversity among breeding materials is what determines the level of segregation and variability among the crossprogeny on which selection is performed. Thus, a well understanding of the genetic diversity among thepotential cross parents, which can optimize the choice of parents to cross and improve the efficiency ofsugarcane breeding, is fundamental to the sucesses of the breeding program. In the present study, a recentlydescribed molecular method, namely target region amplification polymorhpism (TRAP), which preferentiallygenerates polymorphic markers around targeted candidate gene sequences, was applied. Seventy-five mostimportant sugarcane varieties cultivated in Mainland China were selected of 350 sugarcane accessions and wereprofiled using fourteen TRAP primer pairs. In total, these primer combinations produced a total of 144 bands,out of which 103 (70.10%) were polymorhpic. The number of bands amplified from sugarcane samples usingeach primer combination ranged from 3 to 13, with an average of 7.4 bands per primer pair. The polymorphisminformation content (PIC) ranged from 0.8361 to 0.9215 in 75 sugarcane clones and the average was 0.8421.The result showed that the genetic similarity index of 75 sugarcane clones ranged from 0.5138 to 0.9236. It alsoindicated that some sugarcane varieties were abundant in the genetic similarity with wide genetic base, while theothers had low genetic similarity. Based on TRAP profiles, clustering of genotypes was done using unweighedpair-group method with arithmetic averages clustering algorithm (UPGMA) method and it categorized all of theseventy-five examined representative sugarcane samples into two groups at GS 0.667 with fairly moderatebootstrap support. Besides, those in the second group were further clustered into 7 sub-groups at GS 0.752.These major clusters corresponded with the results from principal component analysis (PCA). Therefore, TRAPis an effective tool for estimating genetic diversity and for analyzing the evolutionary and historical relationshipof sugarcane cultivars at the genomic level. It would be helpful in uncovering the genetic basis of sugarcanegermplasm collections, the selection of which parents to cross in sugarcane breeding program and thusaccelerate the process of sugarcane breeding.S 8.12 - FISH strategy for the genomic projects of potato and tomatoTang XM1, Jan de Boer1, Bai YL 1, Dóra Szinay1,2, Lang CT1, Myriam Olortegui Guzman2, Herman van Eck1,3Christian Bachem1, Erwin Datema4, and Roeland van Ham4, Richard Visser1,3, Hans de Jong2 192
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京1 Wagening UR Plant Breeding, Wageningen University and Research Centre; 2Laboratory of Genetics,Wageningen University; 3Center for BioSystems Genomic (CBSG), the Netherlands; 4Applied Bioinformatics,Plant Research International (PRI), Wageningen University and Research Centre.* Email: xiaomin.tang@wur.nl, hans.dejong@wur.nl, or richard.visser@wur.nlFluorescence in situ hybridization (FISH) technology was used for the ongoing potato and tomato genomicsprojects. Our high-resolution mapping of bacterial artificial chromosomes (BACs) and repeats on meioticpachytene chromosome complements features: 1) FISH of all previously mapped BACs simultaneouslyhybridized on potato chromosome 5 (5-multicolour pooled BAC-FISH) revealed several small gaps of about 1Mb. Cot100 genomic DNA was needed to suppress hybridization of repetitive DNA sequences. These resultswill help to focus on new BACs filling the gaps along this chromosome. 2) FISH was also used to demonstratethe chromosomal distribution of major repeat families. Here we characterized and mapped three repetitiveDNAs on Solanum tuberosum, one tandemly repeated DNA family PGR1 (potato genomic repeat 1), twointerspersed repeats P5 and REP2. 3) With the aim of using available tomato and potato BACs to study thechromosomal co-linearity in the Solanum genus, we developed a cross-species FISH strategy. Cot100 genomicDNA was still needed to suppress repeats in the BACs and hybridization stringency was adapted for thecross-species BAC painting. A hitherto unknown paracentric short arm euchromatin inversion in chromosomearm 6S was detected and gave indications for the existence of other smaller chromosomal rearrangementsbetween potato and tomato genomes.S 8.13 - Relationship between expression of BrANT, BrGRF5, BrAN3 and BrARGOS genes and organ sizecontrol in Chinese cabbageWang FD, Zhang S, Li LB, Li HY, Liu LF, Wang CH, Gao JW*Shandong Key Laboratory of Greenhouse Vegetable Biology, Shandong Branch of National VegetableImprovement Center, Institute of Vegetables, Shandong Academy of Agricultural Sciences, 250100 Jinan,Shandong, China.Email: jianweigao3@yahoo.comChinese cabbage (Brassica rapa L. ssp. pekinensis) is a common and economically important crop in Asia. Toobtain information on yield improvement applicable for the genetic engineering approach, we isolated putativehomologs of ANT, GRF5, AN3 and ARGOS from Chinese cabbage genomic DNA, which we designated BrANT,BrGRF5, BrAN3 and BrARGOS, respectively. We first analyzed the genomic structure of BrANT, BrGRF5,BrAN3 and BrARGOS genes. The results indicated that they all contain several introns in their genomicsequence except BrARGOS gene, and in their promoter domain exist many kinds of cis-elements, such asABRELATERD1, ARFAT, AUXRETGA1GMGH3, ERELEE4, WRKY71OS and so on, suggesting that theirexpression could be regulated by various factors. Subsequently, the expression patterns of BrANT, BrGRF5 andBrAN3 in Chinese cabbage were investigated with BrARGOS as positive control. The expression profiles ofBrANT, BrGRF5 and BrAN3 were similar to BrARGOS that they are all have more transcripts in the inbred lineswith bigger heading leaves than the inbred lines with smaller heading leaves and they also have higherexpression levels in the tissue or development stage with stronger ability of cell division. In addition, theexpression of BrANT, BrGRF5, BrAN3 and BrARGOS all could be promoted by NAA treatment and supressed 193
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京by ABA treatment. These results suggest that BrANT, BrGRF5, BrAN3 together with BrARGOS genes mayfunction as some regulators of organ size in Chinese cabbage by enhancing cell proliferation.S 8.14 - Diminution of nutrition inhibitor contents from soybean feed of livestock through RNAinterference manipulationWang PW , Ma J , Fu YP ,Wei YF , Qu J , Zhang J, Yao D , GuanSY and Zhou HTBiotechnology Center of Jilin Agricultural University, Changchun, 130118, China,* Email: peiwuw@yahoo.com.cnTrypsin inhibitor, lipoxygenase and lectin from soybean (Glycine max [L.] Merrill) are main nutrition inhibitorsin livestock feed, leading to a decrease in meat production in raw soybean feeding monogastric animals, such aschicks and swine. Efforts have thus been made over the last decades in diminution of those inhibitive factorsfrom soybean. For example, heating treatment during soybean-products processing was used to reduce theinhibitors. But, it simultaneously declined the nutritional contents of soybeans. Traditional soybean breeding isapplied effectively for elimination of the inhibitive contents from soybean. But this is succeeded only in limitedgenotypes.In this study, we describe a successful diminution of the above-mentioned nutritional inhibitors in soybeanthrough RNAi technique. The selected coding regions of trypsin inhibitor, lipoxygenase and lectin genes wereseparately cloned into an RNAi expression vector harboring a seed-specific promoter and, then, transferredrespectively into a recipient varieties of “jinong18”, “tongnong13” and “jinong27” by Agrobacterium mediatedapproach or pollen tube pathway method. The resulting transgenic plants were examined for both the transgenicexpression and the reductive effects on the content and/or activity of those inhibitors in the transgenic soybean.The results were as follows:1. In the transgenic plants, the mRNA contents of trypsin inhibitor, lipoxygenase and lectin genes weredecreased obviously in the embryos 30 days after pollination in comparison with the non-transgenic controls.This indicated that the gene transcription of trypsin inhibitor, lipoxygenase and lectin in the transgenic plantswere effectively suppressed;2. The enzymatic activities of trypsin inhibitor, lipoxygenase and lectin in the transgenic plant seeds were alsodeclined obviously. In T1 seeds, the activity of trypsin inhibitors was approximately 70% lower than the controland, the activity of lipoxygenase and lectin were both measured to be about 60%-40% lower than the control;3. We examined the contents of protein and oil in T1 seeds using near-infrared spectroscopy. The results showedthat the oil contents of the three transgenic plants increased to different extent in comparison with theircorresponding control. However, their protein contents decreased in all three transgenic soybeans;4. There were no significant changes among agronomic characters in T1 plants in comparison with the controlplants, implying that, as a consequence of RNA interference, the diminution of trypsin inhibitors, lipoxygenaseand lectin in the T1 seeds apparently exhibited no effects on the growth of plant.The above results suggested that RNAi technology may serve as a effective approach in inhibiting the geneexpression of trypsin inhibitors, lipoxygenase and lectin in soybean seed and, ultimately improving the nutritionquality of soybean in a genotype-insensitive manner. 194
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京S 8.15 - A highly efficient method for construction of rice artificial microRNA vectorsWang XM ,Yang Y , Yu CL , Zhou J , Cheng Y , Yan CQ and Chen JP*The Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou, ShiqiaoRoad 198, 310021, P.R.China.* Email: jpchen2001@yahoo.com.cnArtificial microRNA (amiRNA) has become a powerful tool for gene silencing in plants. A new method for easyand rapid construction of rice artificial miRNA vector is developed. The procedure involved modification of thepCAMBIA1300-UR vector by insertion of a ‘vector modification fragment’. This was prepared from theprecursor of Os-amiR528 by eliminating the central miRNA-containing region while simultaneously creating anAfeI restriction site. The fragment was then introduced to the destination vector to produce a multipurpose“Highly Efficient gene Silencing Compatible vector” (HESC-vector). AfeI was used to produce linearizedHESC-vectors, and a blunt end PCR product that included the test amiRNA sequence was cloned into this siteby a single ligation reaction to create the completed amiRNA vector. The method was highly efficient, greatlyreduced the time needed for vector construction and resulted in a DNA sequence identical to that of the currentmethod, making it particularly suitable for use in a systems biology approach to functional genomic research.This method not only reduces the cost of vector construction but also makes it possible for a single member oflaboratory staff to produce numbers of amiRNA vectors simultaneously and is therefore suitable for multiplegene modification. In Arabidopsis thaliana, an amiRNA library has already been set up to determine thefunction of all genes (http://2010.cshl.edu/), providing an example of using a systems biology approach forfunctional genomics research. The same strategy could also be used for gene functional research in rice. Ourmethod can greatly simplify the experimental steps and produce a large number of amiRNA vectors in a shorttime, minimizing time and cost. These outstanding features make it suitable for large-scale gene modificationprojects in rice.S 8.16 - DNA Fingerprinting Technique of Juncus effusus L.Wang ZHInstitute of Biotechnology, Zhejiang Wanli University, Ningbo, 315100Juncus effusus L. var. Decipiens Buchen is Juncaceae rushes of the genus of perennial herbs, commonly knownas Mat grass. The stems can be woven into “TaTaMi” mats, straw mats, straw, grass and grass baskets and otherfans, which have become one of main agricultural products of Ningbo foreign trade income. But since Japanpromulgated "Seed modified laws" on July 8, 2003, many of our products exported into Japan would likelybecome a "violation target" of Japanese patent of new varieties. Therefore, the molecular identification forspecies of our existing rush resources is very important. In addition, in recent years, rush varieties in China wereseriously mixed and deteriorated and greatly affected the production of rush. As the morphological observationis difficult to distinguish the difference among these rush varieties, it is extremely necessary to carry out DNAfingerprinting analysis for rush varieties.Four Juncus effusus L. varieties Yinlin 1, Nonglin 4, Gangshan and Taicao were used to develop DNAfingerprinting technology. All SSR primers were used to perform PCR amplification of genomic DNA in MatRush. The results showed sixty seven pairs of SSR primers can amplify PCR products in four cultivars. The 195
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京result also demonstrated that PCR products amplified by SSR primer RM24 in Juncus effusus L. variety Yinlin 1were different from those in other three varieties, while SSR primers RM267 and RM330 could differeniateNonglin 4 from other three cultivars. Since Yinlin 1 was developed by the breeders in China, RM24 couldprelimarily act as DNA fingerprinting marker to distinguish the varieties of Mat Rush produced by China fromthose produced by other countries and regions.S 8.17 - Cloning and characterization of CcLFY: a LFY homologue in Carya cathayensisWang ZJ, Chen FF, Huang YJ, Zheng BS and Huang JQ *Zhejiang Agriculture and Forestry University, A Nurturing Station for the State Key Laboratory of SubtropicalSilviculture,Linan, Zhejiang Province 311300 People’s Republic of China;*Email: Huangjq@zjfc.edu.cnLFY is a key integrator in floral inductive pathways. As a floral meristem identity, it switches on thedownstream genes that control floral organogenesis. A homologue of LFY, presumed as Carya cathayensis SargCcLFY, was cloned from hickory (Carya cathayensis Sarg.). The cloned cDNA sequence of CcLFY from Caryacathayensis Sarg has 1442 bps which encode a 385 amino acids protein. Structure analysis of the CcLFY genesequence showed that CcLFY has a Pro- rich region, a central acidic domain, a putative Leucine zipper, and abasic region formed by a core of Arg and Lys residues, which are critical motifs of transcription factors (TF).CcLFY protein sequence shares 89.2%, 83.7%, 83.6% and 65.6% identity with Castanea mollissi, Populusbalsamifera subsp., Tricho carpa, Salix discolor and Arabidopsis thaliana, respectively. Southern blot analysisdemonstrated that CcLFY has only a single copy in the hickory genome. It was strongly expressed in bud whileits expression was much weak in mature leaf and stem, and no expression in roots. During floral transition inspring, CcLFY was turned on at early stage, continued increasing then reached the maximum expression aroundMarch 18th. At this stage the main floral bud has been formed. After the expression peak, CcLFY showed anobvious decreasing pattern until the lateral buds formation. In situ hybridization found that CcLFY expressed inboth immature leaf and shoot apical meristem (SAM). During floral organogenesis, CcFLY expression showed adynamic increase pattern from immature leaf to meristem and then floral primordial and bracts, but no anyexpression in mature floral organs. That suggests that the gradual increasing level of CcLFY might be animportant determinant in flower initiation, it also participates the development of floral organ and induces lateralbud by vascular bundle.S 8.18 - Statistical method for mapping QTL of complex traits based on two double backcross populationsXu HM 1, Liu HL 2, Zhu ZH 1 and Zhu SJ11 The agronomy department of Zhejiang University, Hangzhou, 310029, China.2 The maize research institute of Si-chuan agriculture university, Ya-an,625014, China.* Email: hmxu@zju.edu.cn and shjzhu@zju.edu.cnMost important agronomic and quality traits of crops are quantitative, and their genetic variations are usuallycontrolled by a set of quantitative trait loci (QTL) with interaction involved. It is crucial to understand geneticarchitecture of complex trait for designing a efficient breeding strategy of plant breeding. In this research, weintroduced a new experiment design of QTL mapping and corresponding genetic analysis method. The mapping 196
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京population is formed by two backcross populations which is developed by F1 mating with homozygous parentalline P1 and P2 separately. This mapping population allows cross-environment replications, and can be used toanalyze the dominance effects as well as the epistatic effects and QTL-environment interaction, offsettingdrawbacks of a single backcross population. Genetic analysis method based on mixed linear model approacheswere proposed for estimating QTL position and QTL effects including the QTL main effects (the additive effect,the dominance effect, the epistatic effect) and QTL-environment interaction effects (QE). Monte Carlosimulations showed that the proposed method could provide unbiased estimations for both positions and geneticeffects of QTLs. Additionally, the method also exhibited high accuracy and efficiency.S 8.19 - Assessment of genetic diversity and relationship among a collection of CN sugarcane germplasmby SRAP markersXu LP *, Song XX, Que YX and Chen RKKey Laboratory of Sugarcane Genetic Improvement, Ministry of Agriculture, Fujian Agriculture and ForestryUniversity, Fuzhou 350002, China.*Email: xlpmail@yahoo.com.cnGenetic diversity in sugarcane germplasm is the base for the production of hybrids by controlled pollinationtechniques. It is important for the conservation, management and utilization of genotypes and indeed genes inthe breeding gene pool. Compared with morphological and cytological characterization, molecular marker willprovide more accuracy in assessing the extent of genetic diversity and genotyping of the breeding materials. Theobjectives of this study was to assess the diversity of, and establish the relationship between, 109 sugarcanevarieties cultivars and its related generas cultivated in Mainland China. SRAP amplification using 12 primercombinations resulted in a total of 162 scorable markers of which 151 were polymorphic and consisted of93.21%, suggesting that SRAP is a powerful marker technique for fingerprinting sugarcane germplasm. Thepolymorphism information content (PIC) based on SRAPs ranged from 0.8361 to 0.9215 with a mean of 0.8666.The number of alleles per SRAP primer pairs ranged from 8 to 17 and the average was 12.6. Genetic diversityanalysis showed that the genetic similarity index (GS) of 109 sugarcane clones and its related generas rangedfrom 0.5191 to 0.9847. It indicated a high percentage of genetic similarity among the sugarcane varietiesanalyzed and suggested that the sugarcane germplasm collection in Mainland China present a geneticallynarrow base, while much wider genetic distance existed between these sugarcane varieties and its relatedgeneras. Thus, these related generas assessed in this study should be potentially important source of diversityexpansion in sugarcane breeding program. Cluster analysis with UPGMA method and principal componentanalysis (PCR) both grouped these 109 sugarcane geremplasm into two classes at GS 0.635, while the secondclass were further categorized into two sub-calsses at GS 0.765. These results are useful to sugarcane breedersand would promote the utilization of these sugarcane germplasm. It will also help to direct the selection of thecross parents and reasonable distribution of cultivars in Mainland China.S 8.20 - Identification of the Specific SRAP Marker Associated With Cytoplasmic Male Sterility Gene ofBroccoliYao XQ 1 ,Li Y 1 2 , Xie ZJ 1* and Liu LW 2 ,1 Horticultural Research Institute, Shanghai Academy of Agriltural Sciences, Shanghai Key Laboratory of 197
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Protected Horticultural Technology, Shanghai 201106, China;2 College of Horticulture, Nanjing Agricultural University, Nanjing 210095, China.* Email: xiezj8@163.comSRAP (Sequence-Related Amplified Polymorphis) was analyzed between Ogura cytoplasmic male sterility lineXY and its maintainer line XYF of Broccoli.Total 146 pairs of SRAP primers were used. The amplification ofeight primers was polymorphic in the two lines. Only the Me6+Em7 primer had no changed-plant byindividual plant testing. And a 689-bp specific band M6E7-700 was detected in male sterility line but not in themaintainer line. Analysis of the sequence showed that this fragment had no homology with all of the malesterility genes which had been reported,and part of this fragment was highly homologous with part of BACclone KBrB042N05 and KBrB041L12 sequences in Brassica rapa. This suggested that the newly detectedfragment related to cytoplasmic male sterility and properly came from nuclear DNA. This study first discoveredit is different in nuclear DNA between Ogura-CMS line and its maintainer in the Brassica crop. After cloningand sequencing, specific primers were designed to transform the SRAP marker to more stable SCAR marker,which was named M6E7700.The results here offer new clues for supporting the molecular mechanism ofcytoplasic male sterility by cytoplasm-nuclear interaction.S 8.21 - Studies on rot soft resistant gene transformation of AmorphophallusYue Chaoyin1 Guo Zhenghong 1 Wang Jian 1 Liao Tiantian 1 Dai Jianwu 12 Chen Fan 12 HuangYinghua 31 Biotechnology Research Center of China Three Gorges University, Yichang, Hubei, 443002, China;2 Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, 100101, China;3 Department of Botany, Oklahoma State University / USDA-ARS Plant Science Research Laboratory,Stillwater, OK 74075, USA.* Email: yuechaoyin@163.comThe main compound of Amorphophallus konjac was glucomannan (about 30-75%), which has higher medicinaland nutrition value and bright market prospects. Now, the soft rot of A. konjac has been the most obstacles fordevelopment of A. konjac industry. So far, there are no effective measures to control this disease. In this study, aplant expression vector carrying a soft rot resistent gene --AHL Lactonase gene driven by CaMV 35S + ubidouble promoter was constructed and a optimized Agrobacterium based transformation system of A. konjac wasestablished. Callus of A. konjac were regenerated from bulbs placed on MS supplemented with 1.5 mg/L 6-BAand 0.15 mg/L NAA. After pre-cultured for 2 days, the callus of A. konjac were infected for 30min in a solutionof A. tumefaciens with 1.0 OD600 and 100µmol/L AS, and then co-cultured for 2 days. A. tumefaciens wasinhibited with 100 mg/L Kan, and transgenic tissues were cultured on the regeneration medium supplied with0.5g/L activated carbon. Finally the transgenic plants have been obtained with a transformation efficiency of30%. The result of GUS histochemistry stain showed that the heterologous gene could express in the rhizomesand roots of the transgenic A. konjac plants. The transgenic A. konjac plants growen well and showed resistanceto soft rot of A. konjac. This system would lay the foundation for establishing of the transgenic A. konjac withhigh activity against the soft rot of A. konjac. 198
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京S 8.22 - Population structure and genetic diversity in a rice (Oryza Sativa L.) core collection populationinvestigated with SSR markersZhang P, Liu XD, Li JQ *, Li XL and Lu YG *Guangdong provincial key laboratory of plant molecular breeding, South China Agricultural University,Guangzhou, Guangdong, 510642, China.* Email: lijinquan@scau.edu.cn; yglu@scau.edu.cnAssociation analysis has become a promising approach for the dissection of complex traits in plants. Theassessment of genetic diversity and population structure is essential for the efficient organization of breedingmaterial as well as association mapping. Association mapping based on the core collection population wouldmake use of genetic diversity and might enhance the power to detect QTLs. Therefore, the objective of thisstudy were to (1) examine the population structure of a rice core collection; (2) investigate the genetic diversitywithin and among subgroups of the rice core collection; (3) identify the extent of linkage disequilibrium (LD) ofthe rice core collection. A rice core collection consisting of 150 accessions which was established from 2260accessions of Ting’s collection of rice germplasm (consisting of 7128 accessions in total) were genotyped with274 SSR markers and used in this study. Two distinct subgroups (i.e. SG 1 and SG 2) were detected within theentire population by STRUCTURE, principal component analysis and principal coordinate analysis. SG 1consists of 112 varieties including all indica varieties and a few japonica varieties. SG 2 consists of 21 varietiesof typical japonica rice. In addition, 17 intermediate japonica varieties are in the mixed. Further independentSTRUCTURE run on SG1 indicates that SG 1 can be subdivided into four sub-subgroups (i.e. SG 1a-SG 1d),which consist of 24, 35, 21 and 32 accessions, and correspond to intermediate indica, sub-tropical japonica, lateseasonal indica and early seasonal indica, respectively. This hierarchical population structure was confirmed byits neighbor-joining phylogenetic tree. The pairwise Fst among subgroups and sub-subgroups shows maximumvalue (0.2516) between SG 2 and SG 1c, and minimum value (0.1055) between SG 1b and SG 1c. Similar trendis shown for modified Roger’s distance. The average number of alleles per locus for the entire population is 3.88and gene diversity is 0.54. SG 2 shows lower genetic diversity than those of SG 1 and AD. The percentage ofSSR loci pairs in significant (P < 0.01) LD is 33.9% in the entire population and the ratio of linked to unlinkedloci pairs in LD is 1.08. In the subgroups, the percentage of loci pairs in LD is lower and ranges from 1.3 to8.2%, while the ratio linked to unlinked loci in LD varies between 1.31 and 1.52. In the sub-subgroups, thepercentage of loci pairs in LD ranges from 1.6 to 4.3%, where the ratio of linked to unlinked loci in LD variesbetween 1.44 and 1.85. Across the entire population as well as the subgroups and sub-subgroups, r2 decays withgenetic distance, indicating that linkage is one main cause of LD. The results of this study would providetheoretical basis for association mapping using this rice core collection in future.S 9.01 - Fine mapping and cloning of one gene controlling seed coat color in Brassica napusChen X, Xu JS, Wu ZK, Zhao HY, Yuan F, Wu JSand Liu KD*National Key Laboratory of Crop Genetic Improvement, National Center of Plant Gene Research (Wuhan),Huazhong Agricultural University, Wuhan 430070, China.* Email: kdliu@mail.hzau.edu.cnBecause yellow seeded Brassica napus has higher oil and protein content, lower fibre content than black seeded 199
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京one, it has achieved more and more attentions by the oilseed breeders to satisfy our growing demand on thequality of the colza oil nowadays. But owing to the complex of the genetic mode of yellow seed trait and thegenome structure of brassica napus, the study of its molecular mechanism was weaker than that of otheragronomic traits in oilseed. In this research, we selected yellow seed coat line Y5 and black seed coat line Y47from one DH (doubled haploid) population developed by one pure yellow seeded line No.2127-17 and the blackseeded cultivar ZY821. Using the two DH lines which shared similar genetic background, we generated one F2mapping population. Genetic analysis indicated that the seed coat color was controlled by one single partialdominant gene. Further genetic mapping assay by BSA (bulk segregant analysis) technology revealed that 63AFLP (amplification fragment length polymorphism) markers and 11 SSR (simple sequence repeat) markerswere associated with the seed coat color gene, while it was located on the C08 chromosome primarily. Genotypeanalysis of the F2 segregation population containing 4260 lines was performed utilizing these molecular markersfor fine mapping the gene. As a result, it was delimited within a genetic interval of 0.75cM. This study would behelpful in the gene cloning and functional characterization.S 9.02 - Marker-assisted Breeding of Recessive Genic Male Sterility Lines and their TemporaryMaintainers in Rapeseed (Brassica napus)Dong FM, Hong DF, Liu PW, Zhang YH, Liu T, Wen YP, Dong L, Yang GS*National Key Laboratory of Crop Genetic Improvement, National Center of Rapeseed Improvement (WuhanBranch), Huazhong Agricultural University, Wuhan 430070.* Email: gsyang@mail.hzau.edu.cnThe spontaneous recessive genic male sterility (RGMS) line 9012AB was firstly reported in China and isregarded to have great potential in rapeseed heterosis utilization because of their stable and complete sterility,extensive distribution of restorers, and diverse cytoplasmic sources. In our breeding programs, 9012AB and itstemporary maintainer line T45 were individually utilized as the donor parents for introgression of the malesterility genes ms3 or Rf into the excellent polima cytoplasmic male sterility (CMS) lines (DH206A, DH303A,DH987A and DH 195A-14) and open-pollination (OP) cultivars (Zhongshuang 9, Zheshuang 72, Huyou 15),using a marker assisted selection (MAS) strategy. Massive dominant and co-dominant molecular markerslinked to ms3 and Rf were identified in map-based cloning of these two genes, and they were screened betweenthe donor and the recipient parents. Using those markers displaying stable and repeatable polymorphism forforeground selection, we picked out the individuals heterozygous at developed of both the ms and Rf loci, andanalyzed their genetic backgrounds with a lot of AFLP and SSR markers. After six generations’ selection inthree years, we have obtained three new recessive genic male sterility lines T900AB, ZS9AB and 987AB, alsowith their own homologous temporary maintainer lines. Genotypic analysis showed that the genome of thesterility lines and the temporary maintainer lines comprises above 96% of the recurrent parents DNA. Thepreliminary analysis of agronomy characteristics and quality showed that they have exhibited high similaritieswith their corresponding recurrent parents. The breeding of the other lines were ongoing as well as expected.These results demonstrated that MAS is an effective and convenient approach for breeding of new RGMS linesand their homologous temporary maintainers. 200
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京S 9.03 - Anthocyanin accumulation in cotton (Gossypium hirstum) fiber generated by the maize Lc geneintegrated via Agrobacterium-mediated under lightFan XP1,2, Liu J3, Yang WC3*1 Institute of cotton research, Academy of ShanXi agricultural science, China, 044000. 2 Temasek Life SciencesLaboratory, 1 Research Link, Singapore 117604.3 Key Laboratory of Molecular and Developmental Biology and National Centre for Plant Gene Research(Beijing), Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Datun Road,Beijing 100101, China.* Email: wcyang@genetics.ac.cnBreeding of naturally colored cotton fiber has been hampered by the limited germplasm, an alternative way is touse transgenic approach to create more germplasm for breeding. Here we report our effort to engineeranthocyanin production in cotton. The maize Lc gene, under the control of the constitutive 35S promoter andfiber specific expression promoter ACTIN1, were introduced into cotton through genetic transformation. Ourdata showed that the expression of the Lc gene under 35S promoter in tissues of transgenic cotton checked byRT-PCR, and it is sufficient to trigger the accumulation of anthocyanin in a variety of cell types including culli,somatic embryos, young roots, cotyledons and more pigment appeared in leaves, petioles and stems of matureplants. Floral parts also expressed red color in sepal, anther, stigma, but there was no phenotype or molecularsignal checked by RT-PCR in petal. There was no phenotype in leaf epidermis cells ether. Therefore,anthocyanin accumulation in cotton often displayed cell and tissue-specific patterns. The light triggerexperiment showing that the accumulation of colored anthocyanin in cotton young fibers requires theparticipation of light signaling. The 12 lines transgenic plants were generated by Lc gene under ACTIN1promoter showed that almost no color changed in variety tissues and presented less red color in young fiberunder light. These data indicate that it is feasible to engineer colored fibers through enhance promoter specialand its expression strength in transgenic cotton.S 9.04 - Mapping of Nuclear Male-sterile Genes ms14 using SSR markers in CottonHu BM, Hu Lei, Wang PZ*Xinjiang Condy Seed Science & Technology Development Co. Ltd, Urumqi, 830011.* Email: Condywpz@126.comNuclear male sterility (NMS) is a very important character for cotton breeding and genetics programs.Identification of linked molecular markers with NMS will greatly facilitate breeding for this trait. Among all ofthe nuclear male-sterile genes, only Dong-A (ms14) and ms5ms6 lines were utilized successfully in hybridproduction of cotton. Dong-A (ms14), a recessive NMS line developed from spontaneous mutation in uplandcotton, has been applied most widely and played an increasing role in hybrid cultivar development in China. Anintraspecific F2 population comprised of 180 individual plants was developed by crossing a cultivar of Kang A(Dong-A derived Lines, Gossypium hirsutum L.) to a cultivar of 601588 (Gossypium barbadense L.). Base onthis population the ms14 gene was mapped on chromosome 2 and closely linkaged with BNL3971 (simplesequence repeat markers) within a genetic distance of 16.7 cM. The BNL3971markers can be used for themarker assisted selection in breeding a new inbred line, and provide the foundation for gene isolation by map 201
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京based cloning.S 9.05 - Cytological studies of apomixis in Hickory (Carya cathayensis Sarg.)Huang JQ * , Zhang B , Wang ZJ , Huang YJ and Xia GHZhejiang Agriculture and Forestry University, A Nurturing Station for the State Key Laboratory of SubtropicalSilviculture,Linan, Zhejiang Province 311300 People’s Republic of China.* Email: Huangjq@zjfc.edu.cnHickory(Carya cathayensis) which is an important oil tree with high economic value which bears edible nut in ,China show the tendency of apomictic embryos develop as its polyembryony phenomemon and previousresearch. Up to now, it has not been reported the origin of mature embryos and its mechanism of reproduction.Hickory was identified here as a new apomictic species cytologically by paraffin section and flow cytometrydetection. Developmental pattern of embryo sac formation in hickory represented typical polygonum-type andfemale gametophyte developed from functional megaspore which was selected from tetrad after meiosis ofmegasporocyte. Zygote embryos had not been found in numerous histological researches and embryos alloriginated from nucellar cells. Initial cells of nucellar embryo located both at the micropylar end and chalazalend of embryo sac and presented with cell enlargement and vacuolization inhomogenously. Mature nucellarembryo only developed at the nucellus beak region although bud like nucellar embryo also distributed at thechalazal end of embryo sac in some ovules absence of endosperm development which was abortion later. Ploidyof Endosperm in hickory was identical with leaves of mother tree and was diploid by flow cytometry detection.Development of endosperm in hickory was spontaneous type.S 9.06 - The rice CYP81A6 can be used as a bentazon and sulfonylurea herbicide-resistance transgene anda selection marker geneLiu C, Liu SQ, Wang F, Liu KD*National Key Laboratory of Crop Genetic Improvement, National Center of Plant Gene Research (Wuhan),Huazhong Agricultural University, Wuhan 430070, China.* Email: kdliu@mail.hzau.edu.cnCytochrome P450 (CYP450) genes have been widely used to generate transgenic plants for weed control andphytoremediation. Bentazon and sulfonylureas are two different classes of herbicides that can kill broad-leafweeds in rice field. A CYP450 gene CYP81A6 encoding a monooxygenase has been identified in rice, whichconfers the resistance to bentazon and sulfonylureas. In this study, the CYP81A6 gene was isolated from rice(Oryza sativa L. cv. ‘Nipponbare’) and then introduced into Arabidopsis. Transgenic Arabidopsis showed strongtolerance to bentazon and bensulfuron-methyl (BSM), one of sulfonylurea herbicides. Transgenic plants cantolerate 30-120 mg/L of bentazon in soil whereas wild-type plants showed obvious symptom below aconcentration of 30 mg/L of bentazon. Arabidopsis plants expressing the CYP81A6 gene could germinate andgrow healthily in medium containing 1×10-5 g/L of BSM, whereas the wild-type plants could not survive in thesame conditions. In addition, our results showed that the selection efficiency of CYP81A6 as a selection genewas comparable to that of bar gene in Arabidopsis. Using the CYP81A6 as a selection marker, we also screenedtransgenic Arabidopsis harboring the target gene encoding β-glucosidase. These results demonstrated that the 202
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京CYP81A6 gene can not only be used to produce crop plants resistant to bentazon and sulfonylureas to controlthe weeds, but also serve as a novel selection marker for genetic transformation in some dicot crops.S 9.07 - QTLs mapping for fiber quality traits across multiple generations and environments in UplandcottonSun FD, Zhang JH, Wang SF, Shi YZ , Liu AY, Li JW, Gong JW, Yuan YL *Cotton Research Institute, Chinese Academy of Agricultural Sciences , Key Laboratory of Cotton GeneticImprovement , Ministry of Agriculture , Anyang Henan, China, 455004 .* Email: youluyuan@hotmail.comIdentification of stable QTLs of fiber quality traits in multiple generations and environments could facilitatemarker assisted selection (MAS). In the present study, F2, F2:3 and recombinant inbred lines (RILs, F6:8)populations derived from an Upland cotton cross between 0-153, an Upland cotton germplasm that containsintrogressive G arboreum L. chromosome fragment with excellent fiber quality, and sGK9708, a commercialtransgenic Upland cotton cultivar, were constructed for QTL tagging of fiber quality. A total of 5742 SSR primerpairs was used to screen for polymorphism between the two parents. Linkage maps of F2 and RILs were thusconstructed, containing 155 and 190 loci and with a total map distance of 959.4 cM and 700.9cM respectively.The data of fiber quality of F2 and F2:3 family lines obtained in 2003 and 2004 in Anyang, Henan province andthose of RILs in 2007 in Anyang, Henan Province and 2008 in Anyang, Henan Province, Quzhou, HebeiProvince and Linqing, Shandong Province were used to screen the QTLs in multi generation and environmentsthrough Composite Interval Mapping. A total of 51 QTLs for fiber quality were identified and 10 for fiberstrength, 10 for fiber length, 11 for micronaire, 8 for fiber uniformity and 12 for fiber elongation ratio. 8 out of51 fiber quality QTLs were indentified in F2, F2:3 and RILs simultaneously. 2 QTLs for fiber strength located onC7 and C25 were detected in all the three generations and four environments simultaneously and each QTLexplained 16.67%~ 27.86%, 9.43%~21.36% of phenotypic variation respectively. These stable QTLs of fiberquality traits in multiple generations and environments could be usecd for marker assisted selection (MAS).S 9.08 - Cotton molecular breeding: exploiting genes from Gossypium mustelinumWang BH 1,2, Zhuang ZM 1,2, Liu LM 2, Lubbers Ed 2, May OL 2, Zhang ZS 2,3, Auckland S 2,3, Rong JK 2,3 ,Paterson AH 2,3, Zhang TZ 4, Chee PW 2*1 School of Life Science, Nantong University, Nantong, China;2 Department of Crop and Soil Sciences, University of Georgia, Tifton, GA;3 Plant Genome Mapping Laboratory, University of Georgia, Athens, GA;4 National Key Laboratory of Crop Genetics & Germplasm Enhancement, Cotton Research Institute, NanjingAgricultural University, Nanjing, China.* Email: wchee@uga.eduThe cotton genus Gossypium L. consists of about 45 diploid species and 5 tetraploid species, among which 4species are cultivated, namely G. herbaceum, G. arboreum, G. hirsutum (Upland cotton), and G. barbadense.Upland cotton provides the majority of cotton production in the world. However, the narrow genetic diversityrestricts cotton breeding by limiting the range of valuable traits available for use. Wild species are a broad 203
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京resource to widen the germplasm and improve the traits in cotton cultivars. In this research, a tetraploid wildcotton species, G. mustelinum is utilized to exploit genes from wild cotton species through cotton molecularbreeding.The Brazilian endemic cotton G. mustelinum is the species in the polyploid Gossypium clade that is mostdiverged from G. hirsutum, thus potentially harbors the greatest number of novel alleles which could be usefulfor cotton improvement. In this study, we seek to explore the genome structure of G. mustelinum, to understandthe transmission genetics in crosses between G. mustelinum and cultivated G. hirsutum, and to identifyagriculturally valuable alleles from G. mustelinum. Using an interspecific G. mustelinum by G. hirsutum F2population, we constructed a genetic map consisting of 1,055 loci covering all 26 chromosomes. Comparing thismap to other interspecific cotton maps revealed a number of structural changes in the polyploid Gossypiumgenomes. We further developed an advanced backcross population with 35 independently derived BC3F2families with an average size of 151 individuals for a total of 5,305 progenies. Both genotypic and fiber qualitydata were collected from 21 BC3F2 families and QTL analyses for fiber quality traits were performed.Opportunities in utilizing G. mustelinum as source of novel alleles for the improvement of fiber quality will bediscussed.S 9.09 - 油菜 BnERF56 基因通过乙烯和水杨酸信号途径调控对多种腐生真菌的抗性汪承刚 黄军艳 董彩华 刘胜毅中国农业科学院油料作物研究所 核盘菌的侵染诱发了一系列植物抗性相关基因的表达,通过对油菜被核盘菌侵染的 cDNA 芯片的分析, 我们挑选出一个在侵染后 6 小时显著上调表达的基因, 序列分析发现, 它属于油菜乙烯响应因子(ERF)家族的成员, 称为 BnERF56。相应于 BnERF56 作为转录因子的功能发挥, 亚细胞定位实验表明,BnERF56定位在细胞核。用植物激素乙烯,茉莉酸和水杨酸处理油菜,荧光定量 PCR 检测表明,BnERF56 的表达受乙烯负调控, 水杨酸正调控, 而独立于茉莉酸信号途径。 BnERF56 的超表达拟南芥转基因植株 PDF1.2,PR-1 的表达上调,对腐生真菌核盘菌和灰霉菌的抗性显著增强。抗病相关基因 BnERF56 的鉴定为油菜转基因抗病育种提供了新的基因源。S 9.10 - Genetic Analysis and Improvement of a Recessive Genic Male Sterility in Brassica napusXia SQ, Zu F, Zhu Y, Dun XL, Zhou ZF, Yi B, Wen J, Shen JX, Ma CZ, Tu JX*, Fu TDNational Key Laboratory of Crop Genetic Improvement, National Sub-center of Rapeseed Improvement inWuhan, Huazhong Agricultural University, Wuhan 430070, China.* Email: tujx@mail.hzau.edu.cnHybrid cultivars have been successfully used to increase double-low rapeseed production worldwide. To utilizecrop heterosis, two aspects must be carried out. The first aspect is an effective pollination control system, suchas male sterility. Currently, the main approaches for rapeseed hybrid production in China are cytoplasmic malesterility (CMS) and genic male sterility (GMS). There are several disadvantages of CMS, such as limitedavailability of restorer lines and sensitivity to temperature. On the other hand, GMS especially recessive GMShas advantages, such as rich resources of restorers, stable male sterility, and complete male sterility. Comparedwith CMS, GMS is not as easy to generate a completely male sterile population, and this may limit its 204
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京application in hybrid seed production. Brassica napus lines 7-7365ABC are a recessive epistatic genic malesterile (REGMS) three-line system. The genetic mode of the REGMS lines was studied by molecular markertechnology combined with different segregation experiments. Two population of near-isogenic lines,7-736512AB and 7-7365AC, which correspond to the BnMs4 gene and BnRf gene derived from a homozygoustwo-type line 7-7365AB, respectively, were constructed. Comparison of mapping results, six markers, CNU063,ENA06, sR4047, SC25, SC916, and SSR1, were common in both linkage groups between the BnRf gene andBnMs4 gene. The identical two closest markers flanking the BnMs4 gene and BnRf gene had genetic distancesof 1.0 cM and 2.4 cM, respectively. Both gene were located at the same region in the top of the N7 linkagegroup, and might be multiple alleles. This genetic model was used to drive a completely male sterile populationfor hybrid seeds production. But breeders have to use a 1:1 mixed population of 7-7365AB to reproduce themale sterile line. We therefore propose a new method of reproducing the male sterile line using self-pollinatedseeds generated by heat shock. The seeds were crossed with the maintainer line to drive a completely populationfor hybrid seed production. This method makes REGMS a more efficient and safer tool for rapeseed hybrid seedproduction. The second aspect of heterosis utilization is the genetic difference between two parents. Breedingpractices have shown that the appropriate genetic difference between parents is a positive correlation with theF1 yield. We designed a marker-assisted selection strategy to transfer the target genes into two subgenomic(ArArCcCc) materials, and bred new A and B lines in REGMS system. The combining ability analyses of yeildand yeild-related characters were done in the crosses from the new A line and 10 restorers within a randomblock design. The results demonstrated that the improved sterile line showed positive effects of value in thegeneral combining ability (GCA), and gave a good opportunity to select a powerful hybrid cultivar.S 9.11 - Two major QTLs for oleic acid content on the linkage groups A5 and C5 in rapeseed (Brassicanapus)Yang Y, Yang S, Chen Z, Guan C, Chen S, Liu ZOilseed Crops Institute, Hunan Agricultural University, 410128, Changsha, China.* Email: zsliu48@sohu.comQuality of rapeseed oil depends largely on its fatty acid composition. The major fatty acid of the zero-erucic oilis the oleic acid, a key ingredient responsible for the reduction in blood pressure induced by olive oil. In contrastto the polyunsaturated fatty acids, oleic acid is chemically more stable, which makes its shelf-life longer.Therefore, breeding for high oleic acid content extending 80% is currently a major goal of rapeseedimprovement. Mapping QTLs for the acid content could pave a way for molecular breeding and gene cloning inrapeseed. An F2 segregating population from the cross a high-oleic (85.2%) rapeseed line with a low-oleic(60.6%) cultivar in zero-erucic background was used in this study for construction of genetic map and the QTLmapping of oleic acid content. 189 F2 seeds harvested from a single F1 plant were analyzed for fatty acidcomposition by half-seed method of gas chromatogram (GC) using a part of cotyledon. Their oleic acid contentranged from 61.5% to 86.0% and displayed normal distribution as shown by Normality plots with tests using thesoftware SPSS14.0. It is estimated that oleic acid content is controlled by about 2 genes according toCastle-Wright equation. These F2 seeds were then sown to produce the F2 plants. The genomic DNAs wereextracted individually from these F2 plants and genotyped using 329 simple sequence repeat (SSR) primer pairspolymorphic between both parents. A genetic map consisting of 342 SSR loci and 19 linkage groups was 205
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京constructed using the software Joinmap4.0 with regression mapping algorithm and Kosambi’s function. Thesoftware QTL Cartographer 2.5 with the comprised composite interval mapping (CIM) was used to scan theQTLs for oleic acid content in rapeseed. Two major QTLs for oleic acid content were found, each on the linkagegroups(LGs) A5 and C5, totally explaining 75.5% of the genetic variance of oleic acid content. The QTL on LGA5, more important than one on LG C5, had a LOD score of 56.3 and explained 59.1% of the genetic variance.This QTL corresponds to the FAD2 gene which is located on the same position of the QTL, as reportedpreviously. The QTL on LG C5 had a LOD score of 23.4 and explained only 16.4% of the genetic variance.However, this QTL is a new major QTL discovered in this study for oleic acid content and probably associateswith a FAD2 gene which is located on LG C5.S 9.12 - Inheritance and QTL identification for oleic acid content in rapeseed (Brassica napus L)Zhang JF, Qi CK , Chen S , Chen F , Gu H, Gao JQ and Fu SZInstitute of Industrial Crops, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, Jiangsu, ChinaA high oleic acid content line APL01 was crossed with a low oleic acid content line M083, and six basicgenerations (P1, P2, F1, B1, B2 and F2) derived from this cross were used to analyze the genetic model by usingmajor gene and polygene mixed genetic model. The segregation population of (APL01/M083)BC1F1 and 251molecular markers were used to construct a genetic recombination map of rapeseed, which contained 19 linkagegroups, and WinQtlCart 2.0 was used to screen the QTLs linked to oleic acid content in rapeseed. Resultsshowed that oleic acid content was controlled by two additive-dominance-epistasis major genes andadditive-dominance-epistasis polygene. The additive effects of the two genes were 14.35 and 9.88, thedominance effects were -2.30 and -0.45, respectively. The additive-additive epistasis effects equaled to 7.63,which was the largest in three kinds of epistasis. The heritabilities of major genes were 78.25%-92.56%, andthat of the polygene were 0%-6.42%. Four QTLs, qOA1、qOA8、qOA13-1 and qOA13-2, related to oleic acidcontent in rapeseed were identified. qOA1 was located in the region of m19e21c-A0214Ra142 on linkage groupN1, which could explain 3.29% of the oleic acid content variation in segregation population BC1F1. qOA8 waslocated in the region of m11e37b-A0226Ba267 on N8, and accounted for 9.44% phenotypic variation. qOA13-1was located in the region of A0301Bb398-m18e46 on linkage group N13, and accounted for 18.92% phenotypicvariation. qOA13-2 was located in A0226Ba367-A0226Ra425 on linkage group N13, and accounted for 5.35%phenotypic variation. Among these four QTLs, qOA1、qOA8 and qOA13-1 had positive effects, while qOA13-2had negative effects to the oleic acid content. The effect of qOA8 and qOA13-1 were large, and could beregarded as major gene loci, but the effects of qOA1 and qOA13-2 were relative small, and could be regarded aspolygene loci. qOA8 and qOA13-1 could be used as molecular markers in high oleic acid rapeseed breeding.S10.01 - Quantitative trait loci analysis of stem strength and related traits in soybeanChen HF, Shan ZH, Sha AH, Wu BD, Yang ZL, Zhou R and Zhou XNOil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Key Laboratory of Oil CropBiology of the Ministry of Agriculture, Wuhan, 430062, China.* Email: xazhou@public.wh.hb.cnAbstract: Stem strength is one of the major influencing factors of lodging in soybean [Glycine max (L.) Merr.]. 206
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京To identify quantitative trait loci (QTL) associated with stem strength and related traits in soybean, arecombinant inbred line (RIL) population consisting of 165 lines derived from Zhongdou 29 × Zhongdou 32was used in three years, and a prostrate tester was engaged for testing stem strength. Four traits, stem strength,stem diameter, nodes on main stem, root weight were positively correlated with each other at P<0.01. A linkagemap spanning 1240.7 cM was constructed using 245 SSR (simple sequence repeat) markers and a phenotypicmarker (leaflet shape). With the method of composite interval mapping (CIM), total 46 QTLs associated withthe four traits were identified. One major QTL ss1 for stem strength mapped in the marker interval ofSct_067-Satt207 on linkage group (LG) A2 was detected in three years, which accounted for 9.2-19.3%variation with LOD of 3.1-6.6. Most of the QTLs were clustered, especially on LG A2, C2 and L, and someQTLs were mapped to the same loci. The combination of stem strength and related traits may be used as aselection index for lodging resistance with marker-assisted selection (MAS) for its improvement in soybeanbreeding. The results in present study may provide useful information for further genetic studies on stemstrength of soybean.S 10.02 - Identification of Quantitative Trait Loci (QTLs) associated with salt tolerance during seedlinggrowth in soybeanChen HTA,B, Cui SYA,C, Fu SXA, Gai JYA, and Yu DYA,*A National Center for Soybean Improvement, National Key Laboratory of Crop Genetics and GermplasmEnhancement, Nanjing Agricultural University, Nanjing 210095, China.B The Institute of Vegetable Crops, Jiangsu Academy of Agricultural Sciences, Nanjing, 210014, China.C Jiangsu Yanjiang Institute of Agricultural Sciences, Nantong 226541, China.* Email: dyyu@njau.edu.cnSalt stress is an important factor affecting the growth and development of soybean. The inheritance andexpression of traits associated with salt tolerance during the seedling stage is complex. The present study wasconducted to identify quantitative trait loci (QTL) associated with salt tolerance during seedling growth insoybean. Field and greenhouse experiments were conducted to evaluate 184 recombinant inbred lines (RILs)derived from a cross between Kefeng No.1 and Nannong1138-2 for salt tolerance and QTLs that are associatedwith salt tolerance. The molecular map of this RIL population, covering 2625.9 cM of the genome, convergedinto 24 linkage groups, consisted of 221 SSR markers and 1 disease resistant gene (Rsc-7). QTL mapping wasconducted using WinQTLCart. Eight putative QTLs significantly associated with salt tolerance were identified,one QTL was identified both in field and greenhouse experiments. In the field, salt tolerance was assessed(tolerance rating, TR) visually on a 0 (death) to 5 (unaffected by salt stress) scale; Three QTLs were detected ontwo linkage groups explaining 7.1% to 19.7% of the totalphenotypic variance for salt tolerance. In thegreenhouse, plant survival days (PSD) and percentage of plant survival (PPS) under salt stress were measured.Six QTLs were detected on six linkage groups, and explained 7.8% to 19.2% of total phenotypic variation forsalt tolerance. A major QTL was identified between markers Sat_164 and Sat_358 on linkage group G in boththe field and greenhouse. This QTL qppsN.1 identified in the same location as a salt tolerance QTL previouslyreported in soybean. The detection of new QTLs will provide important information for marker-assistedselection (MAS) and further genetic studies on salt tolerance in soybean. 207
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京S 10.03 - Toward identification and mapping SSR markers linked to fresh seed dormancy in peanut (A.hypogaea L.)Faye I1, Fonceka D2, Tossim AH3, Rami JF2 and Ndoye O 1,31 Institut Sénégalais de Recherches Agricoles, Centre National de Recherches Agronomiques (ISRA/CNRA) BP53 Bambey, Sénégal ;2 Centre de Coopération Internationale en Recherche Agronomique pour le Développement (CIRAD), UMRDéveloppement et Amélioration des Plantes, TA A9613, Avenue Agropolis Montpellier, France ;3 Centre d’Etudes Régional pour l’Amélioration de l’Adaptation à la Sécheresse (CERAAS), BP 3320, Thiès,SénégalIn the Semi-arid tropical (SAT) zone, late rains that frequently occur after peanut maturity cause significant (10to 20%) yield losses and seed quality reduction in non-dormant varieties belonging to the subspecies Fastigiata(i.e. Spanish and Valencia market types). These varieties are largely cultivated in SAT for their earliness (90days) that are well adapted to the shortness of the rainy season. However, these varieties lack fresh seeddormancy and then are prone to in-situ germination. In this study, the inheritance of fresh seed dormancy wasstudied in a F2 population of 76 individuals derived from an intrasubspecific Spanish x Spanish cross. Thesegregation ratio for the dormancy trait in the F2 population fit well with a 3:1 (dormant: non dormant)distribution. That statement gives evidence that the trait is monogenic with dormancy dominant overnon-dormancy. Polymorphism between the parents was estimated using 558 microsatellite makers. Apolymorphism rate of 17% was observed. This level of polymorphism was higher than those previously reportedin peanut cultivated x cultivated cross. This relatively higher percentage polymorphism observed in our study ismost likely a consequence of the screening of long-sized microsatellite markers. Bulk segregation analysis(BSA) was performed on the F2 population to identify SSR markers linked to the trait. Unfortunately, we havenot found a linkage between the dormancy trait and the markers we used. Actually, projection of thepolymorphic markers on our peanut map showed that linkage groups a01, a05, a09, b01 and b10 were notsaturated. Therefore, further screening of molecular markers is needed to find markers linked to the trait. Thegrowing number of public available microsatellite markers together with the increase number of cultivated xcultivated genetic maps published offer new opportunities for gene mapping using intrasubspecific crosses inpeanut.S 10.04 - cDNA-AFLP reveals differentially expressed genes related to floral sex determination incucumberFeng Z, Zhi WQ*, WuT,Zhou XJHorticultural Department, Northeast Agricultural University, 59 Mucai Road, Harbin 150030,China.* Email: qzw303@126.comFloral sex determination is a developmentally regulated process culminating in plants bearing unisexual flowers.The process has been extensively studied in flowering plants, particu- larly in cucumber, a monoecious species,to elaborate physiolo- gical and molecular aspects of floral sex determination. In this investigation, weperformed genome-wide transcriptional profiling of apical tissue of a gynoecious mutant (Csg-G) and themonoecious wild-type(Csg-M) of cucumber in an attempt to isolate genes involved in sex determination, using 208
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京the cDNA-AFLP. Using cDNA-amplified fragment length polymorphism, we detected 81 transcriptderivedfragments showing differential expression between the two genetypes that may be involved in cucumber sexualexpression. The fragments were sequenced and analyzed. The fragments were sequenced and analyzed, andexpression of ten genes encoding known proteins was compared between the two strains by reversetranscriptasepolymerase chain reaction-(RT-PCR). RT-PCR analyses confirmed the differential patternsobserved by cDNA-AFLP in six of those genes.S 10.05 - Establishment of SSR fingerprints of Chinese new pear (Pyrus L.) cultivars and its applicationon cultivar identificationGao Y, Tian LM, Liu FZ, Cao YF*Research Institute of Pomology, CAAS/Key Laboratory of the Use of Fruit Germplasm Resources, Ministry ofAgriculture, Xingcheng 125100, China* Email: yfcaas@263.netThe fluorescent labeling of simple sequence repeat (SSR) detection was used to establish SSR fingerprints of 93new pear (Pyrus L.) cultivars.10 pairs of SSR primers screened amplified 132 alleles, with an average of 13.2alleles per locus. The range of observed heterozygosity was 0.2421~0.8632. The amplified banding patterns of10 loci were 8~44 with an average of 29.7. Each cultivar had its cultivar-specific SSR patterns and can use it as areliable method for cultivar identification. Two pairs of primers (KA16 and CH05d04) with high polymorphismcould distinguish all of the 93 pear new cultivars except for the mutants.S 10.06 - TP-M13-SSR technique and its applications in the studies on the analysis of genetic diversity forpear germplasm resourcesGao Y, Tian LM, Liu FZ, Cao YF*Research Institute of Pomology, CAAS/Key Laboratory of the Use of Fruit Germplasm Resources, Ministry ofAgriculture, Xingcheng 125100,China* Email: yfcaas@263.netAn economical method of simple sequence repeat (SSR) detection was used in studies on genetic diversityanalysis of 25 cultivars of 5 species in which fluorescent labeling of SSR-PCR products was used, with tailedprimer M13 (TP-M13-SSR). Two pairs of primers NH013a and NH007b could distinguish all of the pear cultivars,the identified rate of 5 pairs of primers is 76% on average.The ranges of gene diversity, PIC and locusheterozygosity on 5 SSR locuses were 0.6560~0.83460,0.6398~0.8149 and 0.4348~1.0000 respectively. Thegene diversity and PIC of P. sinkiangensis on average which were 0.7160 and 0.6812 respectively were the lowest ,but those of P.Ussuriensis which were 0.7720 and 0.7379 respectively were the highest.The method has theadvantages of high-throughput, sensitiveness, cost-effectiveness and high accuracy. And it had been used instudies on genetic diversity analysis of pear successfully. Therefore, the TP-M13-SSR automated fluorescentdetection system is suggested to be suitable for studies on genetic diversity analysis of many pear accessions bythe use of a large number of SSR markers. The same as other accessions with large genome size, the TP-M13automated fluorescent detection system can also be attempted. 209
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京S 10.07 - Association mapping of drought adaptation in a reference collection of common bean (Phaseolusvulgaris l.)Godwill S. MakundeCrop Breeding Institute, Department of Research & Specialist Services, Harare, ZimbabweCommon bean (Phaseolus vulgaris L.) is one of the most important legumes in the world where it has impactson agriculture, the environment, human nutrition and health (Graham and Vance, 2003). The crop addsbiodiversity in agriculture through positive roles in crop rotations and intercropping with cereals and many othercrops. Their ability to fix atmospheric nitrogen in the soil (Serraj, 2004) makes them play a significant role inboth structure of ecosystems and sustainable agriculture. Production of common bean is mainly in thedeveloping countries dominated by small holder farmers and in Africa is largely cultivated by women(Broughton et al., 2003). Production of common bean is mainly rain fed world wide. The yield realized in theseareas is only 20 – 30% of the genetic potential of improved varieties (Wortmann et al., 1998) due to an array ofboth biotic and abiotic constraints (Sinclair and Purcell, 2005). An estimate of more than 60% of the commonbean crop grown in developing countries suffers from water stress at some stage of crop growth (Thung and Rao,1999; White and Singh, 1991). Water stress periods are more witnessed in Sub-Saharan Africa than any othercontinent. A yield loss of over 300,000Mg of beans occurs due to drought annually in Africa (Wortmann et al.,1998). Annual world wide yield losses are estimated to be US$500 million (Sharma and Lavanya, 2002).Drought management through supplementary irrigation has been an option to increase realizable yields but veryfew smallholder bean growers have access to irrigation water and equipment due to the prohibitive initial costsand monthly charges. The development of drought-adapted common bean varieties is a practical and economicapproach to minimize crop failure and improve food and nutrition security in bean growing areas (Rao, 2001).The objectives of this study are to: a) identify sources of drought tolerance in CIAT reference collection forfuture breeding programs and or as finished products (b) identify simply inherited markers in close proximity togenes affecting drought tolerance. Marker associations can involve discovery of candidate genes if linkagedisequilibrium is at a short distance (c) improve genetic and physiological understanding of drought tolerance indifferent gene pools of common bean through the genetic and physiological characterization of the CIATreference collection (d) establish the relevance of deep rooting towards high yield under terminal droughtenvironment in a selected few Andean and Mesoamerican genotypes from the reference collection. Preliminaryresults indicate that grain formation and filling is the major yield determent that affect final yield under droughtenvironment and was indicated by number of empty pods and seed per plant. Some genotypes from racesDurango and Nueva Granada 2 were not sensitive to water stress. However, these genotypes do not give highyield under optimum moisture conditions. Race Chile and Peru genotypes were not well adapted to the testingenvironments and gave no or little seed yield. Glasshouse experiment for deep rooting ability showed thatgenotypes under water stress conditions had deeper roots than genotypes growing under optimum waterconditions. This supports that deep rooting is an adaptive trait, induced moisture stress in the soil. However,deep rooting alone does not guarantee high yield as indicated by some Andean genotypes possessing deep rootsbut gave low yield under field experiments. The best bet for drought tolerance is to combine deep rooting andgreater grain filling into one genotype. Studies at molecular level to determine polymorphisms for droughttolerance in common bean will be done. Microsatellites and SNPs will be used. 210
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京S 10.08 - Establishment and Optimization of ISSR Reaction System for F1 Generation of CaucasianClover × White CloverHuang F1, Wang MJ 1, He LJ2, Chen L11 College of Ecology and Environmental Science, Inner Mongolia Agricultural University, Huhhot 010019,China2 College of Agronomy, Inner Mongolia Agricultural University, Huhhot 010019, ChinaE-mail: wangmj_0540@163.comThe F1 generation of Caucasian clover × white clover was subjected to ISSR-PCR to investigate the geneticdiversity and the specific molecular markers linked to specific characters. The optimal ISSR-PCR reactionconditions in F1 generation of Caucasian clover × white clover were established by studying the mainparameters. The results showed that the optimum concentrations of seven reactants in 25µL reaction mixturewere as follows: 15ng genomic DNA, 0.2mmol L-1dNTP, 0.4µmol L-1ISSR primer, 1.0U Taq DNA Polymerase,2.5µL 10×PCR Buffer, 2.0mmol L-1MgCl2. The suitable PCR procedure was one preliminary denaturation at94℃ for 2 min; 30 cycles each involved denaturation at 94℃ for 30s, anneal at 52℃ for 45s, extended at 72℃for 1min, and a final extension at 72℃for 7min, then keep the temperature at 4℃.S 10.09 - Development of Soybean Germplasm Lacking of 7S Globulin α-SubunitLiu SS*, Jiang ZQ, Teng WL, Zhang BB, Ge YJ, Diao GZ, Zheng TH, Zeng R, Wu S, Li WBSoybean Research Institute, Northeast Agricultural University / Key Laboratory of Soybean Biology, Ministryof Education, Harbin 150030, Chinaα-subunit of soybean β-conglycinin is one of the major allergens in soybean products. It is possible to reduce orremove the allergens from soybean by development of mutant lines. In this study, a cross was made between 10major varieties in Heilongjiang province of China and “riB” lacking α-, α-, and 11S acidic subunits in order tobreed soybean variety characterized by α-subunit null. F1 seeds were sown individually and individual plantswere harvested to obtain F2-progeny seeds. Subunit compositions of seeds in F1 and F2 were examined forstorage protein by SDS-PAGE. The results showed that all the F1 seeds obtained from cross contained α- andα-subunits. Five band types of 7S globulin in the F2-progeny were observed including α-null,(α+A1aA1bA2)-null, A3-null, (α+A4)-null, and (α+α)-null types.S 10.10 – Found a new soybean (Glycine max) dry resistance mutant and analysis of agronomic traitsMa J1, Wang PW*1, Zhang Z1, Wei YF1, Fu YP1The Biology Center of JILIN Agricultural University,130118,Changchun China.* Email: winter0106@163.comWe found a dry resistance mutant in an experiment in which we intended to decrease lipoxygenase content insoybean seed by RNAi. The germination test of the mutant progeny indicated that it had higher germinationvigor than control group Laboratory germination percent tests showed mutant/control group (following the same)=5.3/1 after germinated 7 days later (following the same). In addition, the root system of mutant progenydemonstrated significant change. The axial root of mutant progeny was slightly longer than control group; 211
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京specifically, the rate of average axial root length was 1.1/1. The fibrous root system, however, was moredeveloped as shown by the facts that the rate of average fibrous root length was 2.6/1 and the rate of fibrous rootaverage was 1.8/1. Drought test in Laboratory indicated that mutant progeny had high drought resistance andthat drought resistance and root length were positively correlated.Agronomic traits indicated the number oflegume of mutant progeny were more than control group 26%.,and the mutant progeny has higher growthpotential, the growth period was advanced about 5 ds, and the productivity increased 28% than the control groupin normal years; the mutant progeny has higher drought-resistance activity, the growth period was advancedabout 7 ds, and the productivity increased 23% than the control group in drought years. Quality inspectionshowed the total protein of mutant progeny was lower than control group 8.3%, fat was higher than controlgroup 5.3%, and the content of Asp、Arg、Prol had significant deviation.S 10.11 - Enhanced level of methionine in transgenic soybean seeds expressing Arabidopsis cystathionineγ-synthase geneSong S 1,2, Hou W 1, Godo I 3, Wu C 1, Amir R 3, Han T 11 The National Key Facility for Crop Gene Resources and Genetic Improvement (NFCRI), Institute of CropSciences, The Chinese Academy of Agricultural Sciences, 12 Zhongguancun South Street, Beijing 100081,China.2 College of Agriculture, Northeast Agricultural University, Harbin 150030, China. 3Laboratory of Plant Science,Migal Galilee Technology Center, P.O. Box 831, Kiryat Shmona 12100, Israel. S.* Email: hantianfu@hotmail.comSoybean(Glycine max (L.) Merr.) seeds are extensively used as high-protein food or feed for human andlivestock all of the world. However, the low level of sulfur amino acids, methionine and cysteine, limits itsnutritional quality. With the aim of increasing the level of methionine in soybean seeds, methionine-insensitiveform of Arabidopsis cystathionine γ-synthase (AtD-CGS), controls methionine biosynthesis, was expressedunder the control of seed specific promoter in two soybean cultivars, Jilinxiaoli No. 1 and Zigongdongdou. Inthe homozygous transgenic seeds, the level of soluble methionine during the reserve accumulation stage and drymature stage increased 6-fold and 2-fold respectively, compared to the level in the wild type seeds. The totalmethionine content in the transgenic Jilinxiaoli No. 1 seeds, including the protein-bound methionine, increasedto about 16-fold in both stages, compared to the level in the wild type seeds. The phenotype and the germinationrate of the transgenic seeds were as those of the wild type. Since the transgenic soybean seeds expressingAtD-CGS have significantly higher levels of both soluble and protein-bound methionine content, this mayrepresent a model and target system for improving the nutritional quality of other legume crops.S10.12 - Transformation of Soybean by Salt Tolerance Gene of BADH via Agrobacterium-MediationWang PW , Ma J , Fu YP , Qu J ,Zhang J , Yao D and Guan SYBiotechnology Center of Jilin Agricultural University, Changchun, 130118, China,* Email: peiwuw@yahoo.com.cnAs an important source of plant protein and oil for human beings, Soybean (Glycine max) is widely cultivatedaround the world. Soybean is moderate sensitive to salt and its salt-tolerance is low. Soil salinization is an 212
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京essential environmental factor which limited the yield of soybean. In this study, the salt-tolerance gene ofBetaine Aldehyde Dehydrogenase (BADH) cloned from suaeda-heteropterakitog was transferred into a soybeanrecipient variety of Jinong17 by agrobacterium-mediation. After 7 generation selection, a novel germplasm ofhigh salt-tolerance soybean was obtained. Molecular analysis suggested that BADH gene integrated into thegenome as double copies and expressed in the transgenic plants, which can grow normally in high salt stress(the content of NaCl in soil is 0.8%). Compared with the non-transformed receptor plants at the initial stage ofsalt stress, the relative electric conductivity of transgenic plants was 25.5% lower, the high molecules leakagevalue was 26.9% lower, the content of MDA was 33.2% lower, the content of proline was 30.2% higher, theactivity of SOD and BADH were higher than the control. Observations of agronomic characteristics suggestedthat the transgenic soybean lines have higher growth potential and drought resistance, the growth period wasdelayed 6-8 days, and the yield increased up to 30% than the control.S 10.13 - Differentially expressed cDNAs of soybean during infection by Phytophthora sojaeXu P. F a 1, WuJ. J b 1, XueA. G. c, ZhangS. Z a, *, LiW. B a, ChenW. Y d, LvH. Y e, LinS. F f, Li N. H a, Fan S. Ja , Wang X aa Soybean Research Institute, Key Laboratory of Soybean Biology of Chinese Education Ministry, NortheastAgricultural University, Harbin,150030, China; b Soybean Research Institute, Heilongjiang Academy ofAgricultural Sciences, Harbin, 150086, China; c Eastern Cereal and Oilseed Research Centre, Agriculture andAgri-Food Canada, 960 Carling Ave Ottawa, Ontario K1A 0C6, Canada;d Suihua Research Institute,Heilongjiang Academy of Agricultural Sciences, Suihua, 152052, China; e Institute of Genetics andDevelopmental Biology, Chinese Academy of Sciences, Beijing, 100101, China; f College of Agriculture andBiotechnology, China Agricultural University, Beijing, 100094, China.* Email: dnzhshzh@yahoo.com.cnTo elucidate the differential gene expression patterns in soybean (Glycine max (L.) Merrill.) during infection byPhytophthora sojae, a suppressed subtraction hybridization (SSH) cDNA library was constructed with cDNAsfrom soybean cultivar “Suinong-10” treated with sterile distilled water as the driver and cDNAs from thatinoculated with P. sojae as the tester. A total of 2067 recombinant colonies from the SSH library were randomlypicked, amplified, and sequenced. After discarding 312 poor quality expressed sequence tags (EST), 1755 highquality ESTs were assembled and edited to 1384 tentatively unique genes (TUG), in which, 586 showedsignificant homology to known sequences, and 798 had low homology or no match with known sequences. AcDNA microarray containing 307 singletons from the 586 TUGs and 222 singletons from the 798 TUGs wasdeveloped to characterize differentially expressed cDNAs in the SSH library, and eight cDNAs were identifiedto be up-regulated after microarray and then confirmed by Real-time PCR. They were homologous topathogenesis-related protein 10, disease resistance response related protein, pathogen-related protein,phenylalanine ammonia-lyase, isoflavone reductase, WRKY transcription factor 31, major allergen Pru ar 1, andpleiotropic drug resistance protein 12, respectively. Most of the up-regulated cDNAs encode enzymes ofphytoalexin biosynthesis and pathogenesis-related proteins to prevent against attack of P. sojae. To ourknowledge, Pru ar 1 is the gene reported for the first time in soybean. The findings of this research contributedto a better understanding of soybean resistance to P. sojae at molecular level. 213
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京S 10.14 - Dissection of genetic overlap of drought and low-temperature tolerance QTLs at thegermination stage using backcross introgression lines in soybeanZhang WB 1,2,$, Qiu PC 2,1,$, Jiang HW 1$, Liu CY1, Li CD4, Fan DM 2, Zeng QL 2, Hu GH 1,3,*, Chen QS 2,*1 Land Reclamation Research & Breeding Centre of Heilongjiang, Harbin 150090, China.2 College of Agriculture, Northeast Agricultural University, Harbin 150030, China.3 The National Research Center of Soybean Engineering and Technology, Harbin 150050,China.4 Heilongjiang Academy of Agricultural Sciences Jiamusi Branch, Jiamusi 150030, China.* Email: qshchen@sohu.com; hugh757@vip.126.com,Northeast of China is the main soybean production area, drought and low-temperature tolerance are both mainfactors involved in reducing soybean yield and limiting planting regions, the effective way is to breed cultivarswith drought and low-temperature tolerance. And the cultivars were able to achieve by BILs in this study. BILs,termed isogenic lines (ILs), or near isogenic lines (NILs), or chromosome segment substitution lines (CSSLs),can be accomplished through interspecific crosses, phenotypic selection, and multiple backcrosses. A set of theBC2F3 lines was constructed with Hongfeng 11 as recurrent parent and Harosoy as donor parent, and screened indrought and low-temperature condition at the germination stage. The 25% (w/v) PEG - 6000 solution was set toidentify the tolerance of drought, and 6℃ to identify the tolerance of low-temperature. The transgressivematerials were chose because their germination time was shorter than the recurrent parent, Hongfeng 11 underthe identification-conditions of drought and low-temperature tolerance, respectively. Related QTLs wereobtained by Chi-test and ANOVA analysis with genotypic and phenotypic data, the probability level wasP<0.05. The results show that 18 QTLs of drought tolerance and 23 QTLs of low-temperature tolerance weredetected. Drought and freezing impose osmotic stress on plants. Upon exposure to osmotic stress, plants exhibita wide range of responses at the molecular, cellular and whole plant levels. Therefore, there is no difficult tounderstand that an overlap to a considerable degree of is existed in its resistance mechanism. Among the QTLsdetected, 12 QTLs were correlated with drought and low-temperature tolerance commonly, which showed apartial genetic overlap between drought and low-temperature tolerance at the germination stage in soybean. Thesimilar research on genetic overlap occurred in rice. In the 12 genetic overlap QTLs, Satt253, Satt513, Satt693,Satt240, Satt323, and Satt255 were detected by at least one method for both drought and low-temperaturetolerance. And Satt557, Satt452, Sat_331, Satt338, Satt271, and Satt588 were detected by only one analysismethod. The mechanism of drought and low-temperature tolerance in crops was complicated; two analysismethods were used to detect QTLs in order to improve the reliability of the results. So the loci detected by bothmethods may show close correlation with drought and low-temperature tolerance of the germination stage insoybean, which lay the foundation for fine mapping of genetic overlap QTL and molecular pyramiding breeding.This will play an important role in MAS for development of both drought and low-temperature tolerancevariety.S 10.15 - Identification of QTLs for yield traits in cultivated peanut (Arachis hypogaea L.)Zhang XY 1, 2,Han SY 2,Tang FS 2,Xu J 2,Liu H2,Yan M 2,Dong WZ2,Huang BY 2,Zhu SJ 1*1 College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310029, Zhejiang, China;2 Industrial Crops Research Institute, Henan Academy of Agricultural Sciences, Zhengzhou 450002, Henan,China. 214
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京* Email: shjzhu@zju.edu.cnTotal bearing branches, mature pods per plant, yield, and 100-pod weight comprise the four major yieldcomponent factors of peanuts (Arachis hypogaea L.). Quantitative trait loci (QTLs) analyses for the yieldcomponents of peanuts were developed using recombinant inbred lines (RILs). A set of RILs (F8:9), including215 lines by single seed descent from a cross Zheng 8903 × Yuhua NO. 4, was developed at the experimentalstation of the Henan Academy of Agricultural Sciences, China. Field experiments were performed following acompletely randomized design with two replicates during each year and for each location. Phenotyping of thepopulation was performed after harvesting. Parental genotypes were screened with 1556 simple sequence repeat(SSR) markers, 85 (5.46%) of which were polymorphic. Segregation data obtained for these markers facilitatedthe development of a genetic linkage map, including 18 linkage groups with 61 SSR loci. WinQTL Cartgraphersoftware was used on the genotyping and phenotyping data. In all, 4 QTLs accounting for 5.36%–7.81%phenotypic variation were identified for the yield components based on the data collected at Sanya site (EI) and7 QTLs accounting for 5.13%–11.73% phenotypic variation for yield components were identified based on thedata collected at Zhengzhou site (EII), China. To detect the interaction effects of QTLs and the environment, theQTLNetwork software was used, after which 9 QTLs accounting for 3.70%–10.54% phenotypic variation foryield components were identified in two environments. These markers should be useful for introgression of themajor QTLs for yield traits of the peanut through marker-assisted selection.S 11.01 - Irradiation mutant mapping of wild beet translocation lines carrying resistance genes againstthe beet cyst nematodeCapistrano GGG *, Jäger SC, Harloff H, Jung CPlant Breeding Institute, Olshausenstr. 40, 24098 Kiel, Germany.* Email: g.capistrano@plantbreeding.uni-kiel.deThe beet cyst nematode Heterodera schachtii Schmidt (BCN) is a severe pest in sugar beet (Beta vulgaris L.)and the only sources of complete resistance are the wild species Patellifolia procumbens and the related speciesP. patellaris. Sugar beet translocation lines A906001 and TR363 carrying a translocation of P. procumbenschromosome 1 are resistance to BCNs. The nematode resistance gene Hs1pro-1 had been cloned from thetranslocation line A906001 using a YAC based 1st generation physical map. However TR363 does not carry thisgene. Hs1pro-1 gave complete resistance in a complementation study done with sugar beet hairy roots but onlypartial resistance was found in whole sugar beet plants transformed using this gene. These were strongindications for a second resistance gene on this translocation. Because the whole translocation is excluded fromrecombination, chromosome breakage mutants were produced. Seeds were 400 Gy gamma irradiated to producebreakages within the translocation and thus narrow down to the resistance gene. The mutants were screenedwith translocation specific markers for the identification of lines with smaller translocations. 2,670 seeds wereirradiated and, after screening of 578 M1 offspring with three molecular markers evenly spread around thetranslocation, two mutants were found. These mutants, named TR320 and TR659, are susceptible to the BCNand had lost most of the translocation region. A BAC based physical map was established with a minimal tilingpath encompassing 18 BACs covering 1,484 kb of the translocation region which had been estimated to be1,500 kb in size. The BAC contigs contain only two small gaps. By collinearity analysis between four wild beet 215
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京translocation lines, the region housing the nematode resistance gene could be narrowed down to ca. 180 kb. Thecandidate sequences present on these regions are being investigated.S 11.02 - Transfer of Anti-Herbicide Gene bar into Upland Rice CultivarsGeng PP1, La HG1, Huang DN2 and Wang HQ1*1 Center of Upland Rice , China Agricultural University,Beijing 100193;2 China National Rice Research Institute,Hangzhou 310006, China.*E-mail: wanghuaqi@cau.edu.cnFor high efficient and secure chemical weed-control, the anti-herbicide gene bar is transferred into cultivars ofupland rice Handao297 etc in this study. The transgenic progenies were established by the anti-basta screeningto calli and regenerating plantlets. Further molecular test by PCR amplification, Southern blotting andbasta-resistant field identifications to T0, T1 and T6 indicated that extrogenous gene bar has integrated into thegenomes of Handao297 and varied with 2 to 6 copy number respectively. The most transgenic lines have 2 to 4bar copies and expressed stable basta-resistant but a few lines showed bar gene-silenced. The promisingtransgenic T8 lines of Handao297 have been developed successfully which performed high resistance to 0.1%basta in seedling stage in field condition.S 11.03 - Mapping and deployment of a leaf rust gene from Aegilops speltoides in hard red spring wheatbreedingHumphreys DG, Hiebert C and McCalllum BAgriculture & Agri-Food Canada, Cereal Research Centre, 195 Dafoe Road, Winnipeg, Manitoba, Canada R3T2M9.* Email: gavin.humphreys@agr.gc.caLeaf rust, caused by Puccinia triticina (Eriks.) is a particularly destructive fungal disease in wheat (Triticumaesivum L.; 2n = 6x = AABBDD). Annual epidemics of this disease cost millions of dollars in losses toproducers. Effective and environmentally friendly control of leaf rust is possible through the deployment ofmajor resistance genes; however, it is imperative that these genes do not have detrimental effects on agronomicperformance or end-use quality. Previous research demonstrated that the ‘Thatcher’ isoline (RL6161) whichpurportedly contained a gene (Lr) derived from Aegilops speltoides (2n = 14, SS) gave a high level of leaf rustresistance. Furthermore, RL6161 had improved grain yield and lower lodging scores compared to Thatcherand was similar to Thatcher for grain protein content, hardness and sedimentation volume (Humphreys et al.2003. Proc. Int. Wheat Genet. Sym. p.724-726). RL6161 has since been used to transfer the leaf rust resistancegene into improved hard red spring breeding material. Deployment and stacking of the Lr gene would befacilitated through marker assisted backcrossing. The purpose of this research was to map the resistance geneand to determine whether the size of the Ae. speltoides translocation was modified during development of hardred spring breeding lines. A doubled haploid (DH) mapping population was produced using F1 plants from thecross Thatcher/RL6161. Segregation of leaf rust resistance fitted a single gene ratio (p1:1 = 0.60). Ninety-sixDH lines were used for mapping with simple-sequence repeat (SSR) markers. Markers revealed that the Ae.speltoides translocation in RL6161 was intercalary and covered a region equivalent to approximately 38 cM of 216
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京the wheat genetic map on chromosome 1BL. Recombination between wheat and Ae. speltoides DNA wasobserved under standard meiotic conditions (i.e. without the use of ph1b etc.) in the DH population. Some ofthese recombinants represented individuals with minimized translocation sizes and restored wheat DNA linkedto the Lr gene. Breeding populations were assessed for similar reductions in Ae. Speltoides DNA after severalcycles of meiosis.S 11.04 - Using Marker Assisted Selection Technique as a tool to identify Cassava Mosaic Disease resistantcultivars in first-backcross populationsAdofo- Boateng P1, Akromah R2, Manu-Aduening J 11 CSIR-Crops Research Institute, P.O.Box 3785, Kumasi;2 Kwame Nkrumah University of Science and Technology, Kumasi.* Email: prisboat@yahoo.com;. rakromah@yahoo.co.uk; jmaduening@yahoo.co.ukCassava mosaic disease (CMD), the most important disease of cassava (Manihot esculenta) is a potential threatto Ghana’s cassava production. The disease is embedded in most landraces resulting in low yields. Host plantresistant has been found to be the best control strategy. A breeding programme using genetic hybridisation andMarker Assisted Selection was initiated in 2007 to improve the resistance levels of farmer-preferred landracesand reduce the long breeding cycle for developing improved cassava varieties. Thirty farmer-preferred landraceswere selected and crossed with a high yielding and mosaic resistant cultivar, TME 11, from IITA. The resultantprogenies found to have a level of resistance to the CMD were AW 18, Nyamebekyere 273, NK 43, AW 3, NK26, K 25, Dabodabo, Ahwengyankwa, 674 Debor, Degarti, Agric Bankye, and NK 57. These resultant progenieswere backcrossed to the resistant cultivar. The backcrossed one (BC1) progenies totalling 224 were screenedwith molecular markers that are associated to the CMD 2 gene. DNA was extracted from leaves of the 224 BC1progenies and 13 parents. Two SSR markers (SSR 28 and NS 158) and one SCAR marker RME 1, were used toscreen and select for the resistant BC1 progenies. Marker Assisted Selection (MAS) revealed that 82% of thegenotypes had at least a marker allele for the CMD 2 gene, indicating resistance. The study further revealed thatby using MAS, the breeding cycle of cassava in the generation of varieties could be reduced from 8 years to 2years. The resistant genotypes identified will be evaluated in future breeding work.S 11.05 - Identification of the bcterial blight resistance genes derived from O. meyeriana.Yan CQ 1, Yang Y 1,Wang XM 1, Yu CL1, Zhou GQ 1,2, Zhang WL 2, Zhou J 1, Cheng Y1 and Chen JP 1*1 Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural SciencesMOA and ZhejiangProvincial Key Laboratory of Plant Virology, Hangzhou 310021, China;2 College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua, 321000 P.R.ChinaThe bacterial blight (BB), caused by Xanthomonas oryzae pv. oryzae (Xoo), is one of the most serious diseasesto the world rice product. O. meyeriana, an important wild rice resource, shows a high resistance to bacterialblight pathogens. The excellent traits of O.meyeriana, which is GG genome, can not be introduced intocultivated rice with AA genome by conventional breeding method. Here, by using asymmetric somatichybridization with O.meyeriana as the donor species and O.sative (Dalixiang) as the recipient species , weobtained Y73 , which is one of the progeny lines and shows high resistance to bacterial blight of rice. Cross of 217
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Y73 and susceptible cultivar IR24 was developed for the identification of the bcterial blight resistance gene.Genetic analysis showed that the the bcterial blight resistance was controlled by polygene and the resistancegenes were recessive. According to the genetic mapping , three resistance loci were mapped on three regions ofchromosome 1, chromosome 3 and chromosome 5, respectively.S 11.06 - Molecular cytogenetic analysis of wheat-Elymus repens introgression lines with resistance toFusarium head blightZeng J, Cao W, Fedak G, Hucl P, Yang Y, Xue A and Chi DEastern Cereal and Oilseed Research Centre, Agriculture and Agri-Food Canada, 960 Carling Ave, Building 50,Ottawa, ON, K1A 0C6, Canada. (J.Z;Y.Y.) Triticeae Research Institute of Chengdu Academy of Sciences,Sichuan Agricultural University, Wenjiang, Sichuan, 611130, R. P. of China. (P.H.) Department of PlantSciences, University of Saskatchewan, 51 Campus Drive, Saskatoon, SK, S7N 5A8, Canada.Elymus repens (2n = 6x = 42, StStStStHH) is a hexaploid species, distantly related to bread wheat Triticumaestivum L. em Thell (2n = 6x = 42, AABBDD). As a potential source of resistance to Fusarium head blight(FHB), Elymus repens was crossed to common wheat in order to transfer resistance genes. Severalwheat-Elymus repens introgression lines with high level of resistance to FHB were developed and designatedP1131-1-5, P1142-1-2, P1142-2-9, P1142-2-5, P1142-2-15, P1142-3-B, P1142-3-E and P1142-3-15. Genomic insitu hybridization (GISH) analysis revealed a variation in number and composition of alien chromosomes.Evidence from GISH indicated that P1131-1-5 contained eight St-H translocation chromosomes and fourwheat-St-H terminal translocation chromosomes plus 44 wheat chromosomes. P1142-1-2 contained 56chromosomes with a pair of translocation chromosomes involving St and H genomes, four wheat-St-Htranslocation chromosomes and eight wheat-St-H terminal translocation chromosomes, plus 21 pairs of wheatchromosomes. In P1142-2-9, a pair of translocation chromosomes of E. repens and ten terminal translocationchromosomes involving St-, H- and wheat chromosomes were detected. In P1142-2-5 and P1142-2-15, tenterminal translocation chromosomes were also identified as trigenomic rearrangement chromosomes. Besides,P1142-2-5 (2n = 53) had one terminal translocation chromosome involving segments from both the St and Hgenomes. In lines of P1142-3-E and P1142-3-B, one and two translocation chromosomes were identified aswheat-alien chromatin interchange involving St-, H- and wheat genomes, respectively, while line P1142-3-1-5with 42 chromosomes carried a pair of terminal trigenomic translocation chromosomes. Meiotic analysisindicated that the translocation chromosomes in this line were homologous and genetically stable. Evaluation ofFHB reaction indicated that all the introgression lines have a high level of resistance to FHB. Thesewheat-Elymus repens introgression lines will be useful for development of improved wheat cultivars withresistance to FHB in wheat breeding programs.S 11.07 - GISH Characterization of Two Hexaploid Partial Amphiploids with Resistance to FHB, LeafRust and Stem RustZeng J, Cao W, Fedak G, Sun S, McCallum B, Fetch T, Xue A and Zhou YEastern Cereal and Oilseed Research Centre, Agriculture and Agri-Food Canada, 960 Carling Ave, Building 50,Ottawa, ON, K1A 0C6, Canada. (J.Z., Y.Z.) Triticeae Research Institute of Sichuan Agricultural University, 555Northeast Road, Wenjiang, Sichuan, 611130, China. (S.S.) Institute of Crop Genetics, Shanxi Academy of 218
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Agricultural Sciences, 64 Nongke Beilu, Taiyuan, 030031, China. (B.M.,T.F.).Cereal Research Centre,Agriculture and Agri-Food Canada, 195 Dafoe Road, Winnipeg, MB, R3T 2M9, CanadaThinopyrum intermedium (2n=6x=42, EeEeSt), a wild relative of wheat, is an excellent source of diseaseresistance. Two partial amphiploids, 08-47-50 and 08-53-55, were developed from wide crosses between durumwheat and Th. intermedium. Meiotic analysis indicated that the two lines contained 42 chromosomes and werecytologically stable. Genomic in situ hybridization revealed that the two partial amphiploids carried differentchromosomal compositions. Line 08-47-50 had a chromosome complement of 28DW+6St+4Ee+4Ee-St, whilethe line 08-53-55 had a chromosome complement of 28DW+4St+10Ee-St. Greenhouse evaluation indicated thatthe both lines had a high level of resistance to Fusarium head blight (FHB) with a severity of 6%, compared tothe susceptible check Roblin at 100%. Leaf rust test showed that these two lines were highly resistant at theseedling stage to the Canadian western co-op trial screening races: MBRJ, MGBJ, TJBJ, TDBG and MBDS andepidemic race mixtures from 2006, 2007 and 2009, which each represented over 50 races. In addition, the twolines were also highly resistant to the most virulent stem rust Ug99 races TTKSK and TTKST. These two partialamphiploids developed in this study could be used as a new source of multiple disease resistance in bread wheatand durum wheat breeding programs.S 11.08 Study on Virus-resistance of Papaya Ringspot Virus Mediated by Double-Strand RNAZhang F, Jiang LCollege of the Department of Horticulture and Forestry of Huazhong Agricultural University, Ministry ofEducation Key Laboratory of Plant Biology, National Indoor Conservation Center of Virus-free Gemplasms ofFruit Crops, Wuhan, 430070In this study, tobacco, papaya and arabidopsis were used as the plant materials.The main experiment methodwas agrobacterium mediated gene transformation. The agrobacterium were harboured pHellsgate12-CPIR, whichwas structured the CP gene inverted repeats. And we constructed the agrobacterium transient expressionexpriment in papaya. The transgenic plants were inoculated with PRSV and then we analyzed thevirus-resistance. After 3 to 7 days, the leaves of WT plants displaying the phenomenon of lesion, wrinkle andchlorosis. And then we carried out the RT-PCR expriments. The expression of CP mRNA was detected in theWT plants infecting with PRSV, but not detected in the transformed plants. It seems that the transformed plantsinduced the RNAi process to resist the PRSV.S 11.09 - Cloning and functional characterization of a NBS-LRR that participates in wheat defenseresponse to BYDV infectionZhao D 1,2 , Li N 1 , Huang X 1,2 , Zhao J 1 , Du LP 1,2 , Huang ZJ 2 and Zhang ZY 1*1 National Key Facility for Crop Gene Resources and Genetic Improvement / Institute of Crop Sciences,Chinese Academy of Agricultural Sciences, Beijing 100081, China;2 Biology College, Hebei Normal Unversity, ShiJiaZhuang050016, China.* Email: zhangzy@mail.caas.net.cnMany plants contain nucleotide-binding domain and leucine-rich repeat (NB-LRR)- type immune receptor that 219
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京function during defense against pathogens. Barley yellow dwarf virus (BYDV) causes significant yield losses ofwheat worldwide. No resistance gene to BYDV has been found in common wheat. Thinopyrum intermedium, awheat wild relative, shows highly resistant to BYDV infection. The distal segment of Thinopyrum intermedium7Ai#1 chromosome long arm carries a BYDV resistance gene (Bdv2), and has been introgressed into wheat-T.intermedium translocation lines. In this study, a full-length cDNA encoding NB-LRR protein namely TNBL1was isolated from wheat-T. intermedium translocation line YW642 based on a differentially-expressing fragmentidentified by cDNA-AFLP analysis. The deduced protein consists of 1059 amino acid residues, and contains acoiled-coil (CC), and a NBS domain, and LRRs. Sequence alignment and phylogenetic analysis indicated thatthe protein TNBL1 is a new member of NB-LRR family. Like active-functional resistance protein MLA inbarley, subcellular localizations of TNBL1 were the cytoplasm and nucleus, evidenced by transient expressionof yellow flurenscence protein fused with TNBL1 in onion epidermal cells. To investigate if the gene TNBL1plays roles in wheat defense responses to BYDV, we generated transgenic wheat plants with overexpressing orRNA interfering of the gene TNBL1. In the loss-of-function experiments, compared to the resistantrecipient-parent, wild-type YW642, the expression levels of TNBL1, SGT1 and RAR1 in these TNBL1 RNAiplants were reduced, importantly, the knocked-down of TNBL1 transcript led to reduced resistance to BYDVinfection. In the gain-of-function results, compared to the susceptible recipient-parent, wild-type Zhong8601,the expression levels of TNBL1 in the positive transgenic plants were markedly increased, and in turn, thesetransgenic plants overexpressing TNBL1 showed significantly resistant to BYDV. These results suggest thatTNBL1 gene participates positively in defense response to BYDV infection, and SGT1 and RAR1 may berequired for the function of TNBL1 gene.S 11.10 - High resolution mapping of Yd2 gene region in barley using the strategy of comparative mappingZhao YH 1, Wang YF 1, Niu HB 2, Liu LD 1, Hu Q 1 and Li RZ 31 School of life science, Ludong University, Yantai, China;2 National engineering research center for wheat, Zhenzhou, China;3 Department of plant and soil science, University of Kentucky, Lexington, USA.* Email: zyhbob@163.comBarley yellow dwarf virus (BYDV) infects all cereal types and causes serious yield losses worldwide. The Yd2resistance gene in barley has provided an effective and durable resistance to BYDV over 50 years. In an attemptto identify and characterize the Yd2 gene, a map-based cloning strategy has been undertaken in our laboratory.Constructing a high-resolution genetic map of Yd2 region was the first step towards the isolation of Yd2. Thougha linkage map of the Yd2 region of barley chromosome 3 had been constructed and published, the saturation ofthe map was not enough high to fit for map-based cloning. Detecting markers closely linked to Yd2 was a verydifficulty work. The strategy of comparative mapping allowed us to resolve molecular markers previouslyunresolvable from Yd2. To date, many genetic maps of the whole barley genome has been constructed by thediverse mapping projects. Using cMap program, we compared many barley genetic maps previously constructedand developed the consensus map of the Yd2 region. Information generated from the diverse mapping projectswas further enhanced by the development of consensus map. More markers were integrated into the consensusmap, based on the collinearity of genetic maps. Finally, we obtained a putative integrated genetic map of Yd2region with 114 molecular markers using comparative mapping. The high-density map spanned a genetic 220
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京distance of 1.2cM providing an average density of one marker every 0.01cM. Forty-eight markers were closelylinked to Yd2 gene, less than 0.1 cM from Yd2. Among them, six markers were about 0.01 cM from Yd2, theclosest markers. All these markers were involved in SSR, AFLP, RAPD, RFLP, CAPS and STS etc. Wetransformed all RFLPs into STSs for convenient manipulation. The PCR primers of all these markers were listedin this study. This high-density integrative map can provide useful information for map-based cloning of Yd2gene.S 12.01 - Enhancing chickpea production for drought tolerance by using marker assisted backcrossing(MABC) and marker assisted recurrent selection (MARS) approachesChamarthi SK 1, Gaur PM 1, Kumar A 1, Thudi M 1, Tripathi S 2, Krishnamurthy L 1, Kimurto P 3,Chaturvedi SK 4, Eshete M 5, Kileo R 6, Varshney RK 1,7*1 International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Patancheru-502 324,India;2 Indian Agricultural Research Institute (IARI), Pusa Campus, 110 012, New Delhi, India;3 Egerton University (EU), Njoro-20115, Kenya; 4 Indian Institute of Pulses Research (IIPR), Kanpur-208 024,India; 5 Ethiopian Institute of Agricultural Research (EIAR), Addis Ababa-2003, Ethiopia;6 Lake Zone Agricultural Research Institute (LZARI), Mwanza-1433, Tanzania;7 Generation Challenge Programme (GCP), c/o CIMMYT, 06600 Mexico-DF, Mexico.* Email: r.k.varshney@cgiar.orgChickpea (Cicer arietinum L.), is the third most important cool season food legume in the world. It has beengrown extensively by the poor farmers throughout the Indian subcontinent. India alone contributes a share ofabout 67 % to the global chickpea production. However the crop productivity is adversely affected by a numberof abiotic and biotic stresses. Among abiotic stresses, terminal drought is one of the major limiting factors inchickpea production across the world and more than 70% of chickpea growing area is affected by this. With anobjective to develop improved varieties of chickpea with enhanced drought tolerance, two modern breedingapproaches, namely marker-assisted backcrossing (MABC) and marker-assisted recurrent selection (MARS) arebeing deployed in chickpea breeding programme at ICRISAT in collaboration with its partners. In context of thefirst approach (MABC), a hotspot harboring several root trait QTLs with >30% phenotypic variation has beenidentified based on extensive genotyping and phenotyping data from two mapping populations. Genomic regionbracketed by two SSR markers (TAA170, ICCM0249) is being introgressed in three elite chickpea lines (JG 11,Chefe and KAK 2). Above mentioned two SSR markers have been used for foreground selection, while sixAFLP primer combinations have been used to undertake background selection. After completing three cycles ofMABC and two cycles of selfing, BC3F3 plants are being raised at present for seed multiplication. The BC3F4progenies from these plants will be grown for testing their agronomic performance. In the second approach(MARS), favorable alleles for drought tolerance are being pyramided by using two different crosses (JG 11 ×ICCV 04112, JG 130 × ICCV 05107). F3 plants from these crosses are being genotyped and F3:5 progenies arebeing phenotyped under rainfed and irrigated conditions at different locations in India and Africa. QTLs will beidentified based on analysis of F3 genotyping and F3:5 phenotyping data and superior QTLs alleles will beaccumulated by three cycles of recurrent selections. The progenies so developed will be evaluated inmultilocation trials. In summary, it is anticipated that progenies developed following MABC and MARSapproaches may have enhanced drought tolerance. 221
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京S 12.02 - Differential expression of miRNAs in response to salt stress in maize rootsDing DCorn (Zea mays) responds to salt stress via changes in gene expression, metabolism and physiology. Thisadaptation is achieved through the regulation of gene expression at the transcriptional and post-transcriptionallevels. MicroRNAs (miRNAs) have been found to act as key regulating factors of post-transcriptional geneexpression. However, little is known about the role of miRNAs in plants responses to abiotic stresses. A customparaflo microfluidic array containing release version 10.1 plant miRNA probes (http://microrna.sanger.ac.uk/)was used to discover salt stress-responsive miRNAs using the differences in miRNA expression between thesalt-tolerant maize inbred line NC286 and the salt-sensitive maize line Huangzao4. miRNA microarrayhybridization revealed that a total of 98 miRNAs, from 27 plant miRNA families, had significantly alteredexpression after salt treatment. These miRNAs displayed different activities in the salt response, and miRNAsbelonging to the same miRNA family showed the same behaviour. Interestingly, 18 miRNAs were found whichwere only expressed in the salt-tolerant maize line, and 25 miRNAs that showed a delayed regulation pattern inthe salt-sensitive line. A gene model was proposed that showed how miRNAs could regulate the abioticstress-associated process and the gene networks coping with the stress. Salt-responsive miRNAs are involved inthe regulation of metabolic, morphological and physiological adaptations of maize seedlings at thepost-transcriptional level. The miRNA genotype-specific expression model might explain the distinct saltsensitivities between maize lines.S 12.03 - QTL analysis for plant height and heading date in rice under two nitrogen levelsFeng Y , Zhai RR , Cao LY , Lin ZC , Wei XH and Cheng SHChina National Rice Research Institute, Hangzhou 310006, China.* Email: hcheng@mail.hz.zj.cnA recombinant inbred lines (RIL), derived from a super hybrid rice cross of XQZB/R9308 and its geneticlinkage map, were used to identify QTLs for the plant height (PH) and heading date (HD) in rice under N- andN+ conditions. A total of 10 QTLs for PH and 8 for HD were detected under two nitrogen levels, and each QTLexplained 5.68%-18.40% of total variation. The QTL of which contribution rate was 16.99% and 15.85% forplant height was identified at the interval of RM3670-RM2 on chromosome 7 under two nitrogen levels. TheQTL of which contribution rate was 18.40% and 15.70% for heading date was identified at the interval ofRM180-RM5436 on chromosome 7 under two nitrogen levels. One pleiotropic QTL controlling plant height andheading date simultaneously was identified at the interval of RM5556-RM310 on chromosome 8 under twonitrogen levels, which suggested that this chromosomal region may be enriched with the key genes for plantheight and heading date. QTL × N-level interaction were detected for plant height and heading date in rice. Itwas supposed that the QTL only detected at low nitrogen stress might be related with high-efficiency utilizationof nitrogen in rice.S 12.04 - Research on Genetic Variation Mechanism of Diploid and Tetraploid of Siraitia grosvenoriiFu W, Ma XJ 222
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京Institute of Medicinal Plant Development, Chinese Peking Union Medical Colledge, China Academy MedicinalScience, No. 151 Malianwa North Road, Haidian District, Beijing 100193, P. R. China,* Email: fuwei0212@163.comIn 2005, our group found a mutant plant from generations of BoLin and ND in experimental field. This mutantis bigger than any other speies. Its flower is wide particularly, while pollen is abortion seriously.This research was to analysis the karyotype and genetic polymorphism between normal and mutant plant ofSiraitia grosvenorii. Genetic differences and polymorphism ananlysis included the genomes (DNA) and thegene expression level (RNA) by SRAP molecular markers.The results showed that normal plant were diploid with chromosome number is 2n=2x=28, while the mutantplant were tetraploid with chromosome number is 2n=4x=56. Therefore the latter is also called asautopolyploidy.Polymorphism among the different ploidy was analyzed based on SRAP markers. There were about 1010recordable bands generated by 196 primer combination, which averaged 5.2 bands for each primer pair. Theresults generally indicated that the genetic diversity revealed by the SRAP among the diploid and autopolyploidof Siraitia grosvenorii woud be quite low by comparions of other plants.The study was to explore molecular mechanism of differential expression between diploid and tetraploid byusing cDNA-SRAP. About 2150 amplifiedbands were obtained with 196 pairs of SRAP primer, and length ofthem between 100-800bp. Differential fragments were about 1250-1300. Stable differential fragments weresorted out, cloned and sequenced. Sequence analysis revealed that most fragments had significant homologousnucleotide sequence with resistant and photosynthesis genes concluing pyruvate kinase, zinc finger protein,cellulose synthase, proline-rich protein, ribulose-1,5-bisphosphate carboxylase/oxygenase and so on. The resultsindicated that tetraploid of Siraitia grosvenorii has more resistant and photosynthesis ability.S 12.05 - Transcript profiling and identification of molecular markers associated with drought tolerancein TeaGupta S*, Bharalee P, Bhorali R, Gohain T, Bandyopadhyay B, Das SK, Agarwal N, Singh HR, Ahmed P,Bhagwati P, Bhattacharya N, Borchetia S, and Das STea Research Association, Jorhat, Assam, India.* Email: sushmitapakhi4u@rediffmail.comTea (Camellia sinensis), being one of the widely cultivated cash crop in India, is prone to high yield losses dueto recurring droughts. Understanding the genes that govern tea plant architecture and response to drought stressis urgently needed to enhance breeding in tea with improved drought tolerance. In order to identify, develop andimplement MAS in Tea, we took whole genome transcriptome polymorphisms analysis approach providinginsight into drought stress-related gene activity. Whole genome transcript profiling was done using c-DNAAFLP technology. Two genotypes ie., drought tolerant(TV-23) and drought susceptible(S.3/A.3) were selectedbased on physiological data and field performance, an induced drought experiment was performed. c-DNAAFLP based transcript profiling was done at two different stages of induced drought ie., before wilting stage andwilting stage. Based on c-DNA AFLP transcript profiling, we identified approximately 450 transcript DerivedFragments (TDFs) presumed to be involved in drought stress. Out of these we were able to associate 35 TDFs 223
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京with the trait and found to be differentially expressed in tolerant plant and identified as putative candidate genesmarker. These TDFs were selected for EST development to develop gene markers for identification of droughttolerant genotypes in tea. Hierarchial clustering formed two distinct clusters, first cluster distinctly separatesdifferent stages of drought i.e., time point of sampling while the second cluster separates the two germplasm.Functional analysis of TDFs showed genes related to metabolism, energy production, transport and signaltransduction. This study provides the first global catalogue of Tea genes and partial transcripts expressed duringdifferent stages of drought stress, together with their functional annotations. This will help to elucidate themolecular basis of the drought tolerance and identify markers for selection of drought tolerant genotypes in tea.S 12.06 - Evaluation of screening effectiveness on stress tolerance in advanced introgression populationsin riceMeng LJ1, Lin XY2, Sun HB3, Ma XF4, Zhu LH1, Xu JL1*, Li ZK1,5*1 Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081, China;2 Institute of Rice, Jilin Academy of Agricultural Sciences, Changchun, 136100, China;3 Institute of Crop, Tianjin Academy of Agricultural Sciences, Tianjin, 300112, China;4 Institute of Rice, Liaoning Academy of Agricultural Sciences, Shenyang, 110101, China;5 International Rice Research Institute, DAPO Box7777, Manila, Philippines.* Email: xujlcaas@yahoo.com.cn; Lizhk@caas.net.cnUsing BC2F6 advanced backcross populations derived from crosses between Chaoyou1 as recurrent parent and11 diverse entries (eight indica varieties and three japonica varieties) as donor parents, salt tolerance (ST) at theseedling stage and the stage from tillering to mature, cold tolerance (CT) at the booting stage and heat tolerance(HT) at flowering stage were identified and screened under the stress intensity when the check, Chaoyou1completely died or almost infertility under stress. The percentage of ST plants was much difference amongvarious populations at seedling stage and the stage from tillering to mature. The percentage of ST plants wasaveragely 4.3% for the introgression progenies from eight indica donors, three times as high as that of theintrogression progenies from three japonica donors. The percentage of CT plants at booting stage was nodifference of introgression populations between indica and japonica donors. It indicated that in Chaoyou1background indica donors hold stronger genes or epistatic loci which can improve ST at the stage from tilleringto mature.Different patterns of ST at the stages of seedling and from tillering to mature were revealed across populations,for example, X21 showed consistent stronger ST at the two stages, the ST of X22 and Bg90-2 became weak tostronger from seedling to later stage while the opposite was true for Chhomrong, indicating ST between the twostages is genetically independent. The progenies derived from different donors showed different screeningeffectiveness on multiple stress tolerance. More ST and CT individuals were selected in the introgressionprogeny of X22 as a donor, more ST and HT plants selected in the progeny of Bg90-2 and more CT and HTplants selected in the progeny of Chhomrong and Yuanjing7, indicating the screening effectiveness for multiplestress tolerance was evident in the introgression populations from above four donor parents in Chaoyou1background.The average fertility of HT and CT plants selected from the progenies in which Fengaizhan1 and Yuanjing7 asdonor parents was high, meaning these two donors are useful for improvement of HT and CT of Chaoyou1. The 224
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京fertility of CT plants selected from the progenies of OM997, Doddi and Chhomrong as donor parent was above70%, suggesting the three donors are beneficial to improve CT of Chaoyou1. The fertility of ST plants screenedfrom the progenies of X21, Q5, X22, OM997, Shennong265 and Bg90-2 were more than seven times as high asthat of Chaoyou1, suggesting these donors will be ideal to improve ST of the recurrent parent. The resultsindicated that most backcross populations showed segregations of ST, CT and HT even although the donorparents don’t have better stress tolerance themselves, and suggesting most donor parents possess favorablegenes for the tolerance or resistance the stresses‘ hidden genetic variation ’ which are very useful for resistancerice breeding.S 12.07 - QTL Identification and the Genetic Overlap of High Yielding, Drought and Salinity Tolerance inRiceWang WS 1, 2, Guan YS 1, 2, Xu JL 1, Elec V 2, Ali J 2 Zhu LH 1 and Li ZK 1, 2*1 Institute of Crop Sciences / National Key Facility for Crop Gene Resources and Genetic Improvement,Chinese Academy of Agricultural Sciences, Beijing 100081, China;2 International Rice Research Institute, DAPO Box 7777, Metro Manila, Philippines.* Email: lizhk@caas.net.cnRice production is limited by many environmental factors, of which drought and salinity are the most importantabiotic stresses which greatly constrain yield and planting area. Development of drought and salinity tolerant(DT, ST) variety is the most effective method to solve this problem. Two BC1F2 introgression populationsderived from an elite indica variety Huang-Hua-Zhan (HHZ) as the recurrent parent and indica varieties IR64and CDR22 as donor parents, were screened under irrigated, drought and salinity conditions at IRRI. 27 highyielding (HY) ILs, 22 drought tolerance (DT) ILs and 30 salinity tolerance (ST) ILs were selected from the twopopulations. These selected ILs were analyzed using X2 to identify candidate QTLs located in the donorsegments associated with HY, DT and ST. In HHZ/IR64 population, QTLs affecting HY, DT and STrelated-traits detected were 5, 8 and 16 respectively, locating in 10 chromosomes except 10 and 12. InHHZ/CDR22 population, 19 QTLs affecting HY, DT and ST were identified in chromosome 1, 2, 4, 5, 6, 7, 8and 9. Among of those QTLs, 7 contributing to HY, DT and ST, 2 affecting HY and DT, HY and ST, DT and ST,respectively. Genetic overlap regions affecting HY, together with DT and/or ST detected in these twopopulations were concentrated in the distribution of chromosomes 1, 6 and 7. In contrast to the high geneticoverlap among the three traits in HHZ/CDR22, there were few shared loci identified in the HHZ/IR64,indicating the 2 donors contributed different sets of loci to the DT and/or ST in the selected ILs. Our resultsprovided useful information and materials for further improvement of DT and ST in rice by pyramidingbreeding, and for detailed molecular dissection of DT and ST in rice using high throughput genomic tools.S 12.08 - Genetic Dissection of QTL for Leaf Water and Canopy Temperature Characteristics Associatedwith Drought Tolerance in WheatYang DL 1,2, Jing RL 2, Li W 1and Chang XP 21 National Key Facility for Crop Gene Resources and Genetic Improvement / Key Laboratory of CropGermplasm and Biotechnology, Ministry of Agriculture / Institute of Crop Science, Chinese Academy ofAgricultural Sciences, Beijing , 100081, China; 2 College of Life Science and Technology, Gansu Agricultural 225
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京University, Lanzhou, 730070, China.* Email: jingrl@caas.net.cnDoubled haploid lines (DHLs) previously constructed by a cross between Hanxuan 10, a drought tolerantcultivar, and Lumai 14, a high-yield cultivar in well-watered field, and ITMI recombinant inbred lines (RILs)previously obtained by single-seed descent (F9) from the cross between Opata85, a Mexican wheat cultivar, andW7984, an amphi-hexaploid cultivar, were used as experiment materials in this study. Some importantphysiological traits associated with drought tolerance including the leaf relative water content (RWC), the rateof excised-leaf water loss (RWL) and the canopy-air temperature difference (CTD) from two wheat geneticpopulations were investigated in different locations under two water regimes (drought and well-wateredcondition). And, QTL (quantitative trait loci) mapping and QTL × water environment interactions (QEIs) wereanalyzed for these target traits in present study. The detailed results were obtained as follows: Major QTLs forRWL, RWC and CTD were mainly distributed on chromosomes 1D, 2A, 2B, 3B, 3A, 3D, 4A, 5A, 4B, 5D, 6B,6D, 7A and 7B in DHLs and RILs. Some co-located QTLs for RWL were found in specific intervals XGli1~XksuD14.1 on 1D, Xgwm610~Xgwm397 on 4A, Xgwm193~P3470-210 on 6B and XksuG48~Xfba187 on 6D,respectively. And, one major QTL for RWC, QRwc.cgb-2B-2, as well as one major QTL for CTD, QCtd.cgb-4B,were respectively detected in the interval Xcdo678~Xcmwg660 on 2B and Xgwm495~Xgwm149 on 4B in2005 and 2006. Two major QTLs, QRwl.cgb-6B-1 and QRwc.cgb-3B-1 were investigated the significantadditive QEIs (A-QEIs) with rainfed environments, explaining phenotypic variation of 10.55% and 9.15%,respectively. Among epistatic QTLs, one pair for RWC on 2D-7B and two pairs for CTD on 2B-3B and 1D-7Bshowed higher phenotypic variations explaining with 22.15%, 12.67% and 13.19%. Moreover, the epistasis andthe epistatic QEIs (E-QEIs) showed higher phenotypic variations, indicating that epistasis and E-QEIssignificantly affected the inheritances for three target traits.S 12.09 - SSSL-based identification of QTLs for cold tolerance at different growth stages anddevelopment of rice variety with cold tolerance at all growth stages by gene pyramidingYang TF 1, Wang XF 1, Zhang GQ 2, Zhang SH 1*, Huang ZH 1, Zhao JL1 and Liu B 1*1 Rice Research Institute, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China;2 South China Agricultural University, Guangzhou 510641, China.* Email: lbgz_2006@yahoo.com, szhanggz@tom.comCold stress is one of the key abiotic stresses in rice production and may occur at different growth stages.Developing cold-tolerant variety is the most economical and effective way to prevent cold damage. However,breeding for cold tolerance in rice is difficult because of its polygenic nature. Understanding the genetics of coldtolerance and performing molecular breeding is the fundamental way for effective breeding.In this study, twenty-six varieties from 10 countries were used to screen for rice germplasm with cold toleranceat different growth stages. Five varieties with survival percentage above 80.0% under 5℃ for 10 days atbudburst stage, one variety with survival percentage above 78.0% under 11℃ for 7 days at seedling stage andtwo varieties with cold tolerance index over 0.65 under 16℃ for 7 days at heading stage were identified.Interestingly, no variety showed good cold tolerance at all three stages and no significant correlation betweenstages was found. By backcrossing the above cold tolerant varieties to an elite cultivar “Hua-jing-xian74”, a set 226
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京of single segment substitution lines (SSSLs) with cold tolerance at different stage were developed. QTLs forcold tolerance at heading stage and seedling stage were identified using the SSSLs at 16℃ and 11℃ created byphytotron, respectively. At heading stage, seven QTLs were mapped on chromosome 3, 6 and 12. The additiveeffect percentage of individual QTL ranged from 21.4% to 25.5%. At seedling stage, two cold-tolerantindicators, cold tolerance (CT) scored in scale under cold treatment and survival percentage (SP) under normalconditions after cold treatment were used to evaluate the cold tolerance of the SSSLs. Five QTLs for CT weremapped on chromosome 4, 6 and 12. The individual additive effect percentage ranged from 16.4% to 33.3%.Among the five QTLs, two QTLs could be detected in two seasons. Five QTLs for SP were mapped onchromosome 4, 6 and 12. The individual additive effect percentage ranged from 15.8% to 53.1%. Three QTLscould be detected in two seasons. Two QTLs were detected in two seasons using either CT or SP as indicator.Identification of QTLs associated with cold tolerance at budburst stage using the SSSLs is underway andmarker-assisted pyramiding of cold-tolerant genes at different growth stages will be performed to develop lineswith cold tolerance at all stages.Our results suggest that different genetic mechanisms control the cold tolerance at different growth stages in riceand it can be a good strategy to identify the genes for cold tolerance at different growth stages and perform genepyramiding breeding. Since SSSLs have only single chromosomal segment difference with their recurrent parent,it makes identification of QTLs for cold tolerance at different growth stages simpler and more accurate, and theyare particularly useful for gene pyramiding breeding.S 12.10 - Genetic Basis of Drought Tolerance at Seedling Stage in Wheat by Seedling Biomass AnalysisZhang JN 1,2,3, Wei TM 1 , Chang XP 1 , Liu GR 3 and Jing RL 1*1 National Key Facility for Crop Gene Resources and Genetic Improvement / Key Laboratory of CropGermplasm Utilization, Ministry of Agriculture / Institute of Crop Science, Chinese Academy of AgriculturalSciences, Beijing 100081, China;2 Hebei Academy of Agriculture and Forestry Sciences, Shijiazhuang 050031, China;3 College of Agronomy, Agricultural University of Hebei, Baoding 071000, China.* Email: Jingrl@caas.net.cnWheat (Triticum aestivum L.) is the staple food crop in China, but frequently suffers drought stress duringgrowing season. It makes the improvement of drought tolerance (DT) in wheat especially important. In thisstudy, a nature population consisting of 309 accessions was employed as the plant materials to dissect thegenetic bases of drought tolerance at seedling stage by genetic diversity analysis and association analysis. Theplants were grown in the containers and six replications were set to two water regimes (three underdrought-stress (DS) and three under well-water (WW)). The experiment was repeated for three times. Droughttolerance coefficient (DS/WW) of seedling biomass, which was closely related to the early vigor under droughtstress, was used for evaluating drought tolerance. Base on the seedling biomass drought tolerance coefficient(DS/WW), 309 accessions were divided as three classes: high tolerance, medium tolerance, and sensitive todrought, respectively. The accessions were genotyped by 83 SSR markers evenly distributed on 21chromosomes. The results showed that genetic diversity indexes of highly drought-tolerant accessions in Dgenome and 5th homoeologous group were lower than that of sensitive accessions. D genome and 5thhomoeologous group may receive Hitchhiking effect of drought tolerance. A total of 15 loci were detected 227
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京associated with seedling biomass under two water regimes. Xwmc311-7B, Xgwm515-2A were identified underboth water regimes, DS and WW. Xgwm135-1A, Xcfd43-2D associated with seedling drought tolerance.Allele-104 of Xgwm135-1A and Allele-166 of Xcfd43-2D were elite alleles correlated with seedling droughttolerance. These markers may play an important role in wheat molecular breeding for seedling drought-tolerantimprovement.Gene discovery and pre-breeding for Russian Wheat Aphid and Ug99 resistance in wheatM. Cakir1,16, H. Phan1, Janine Vitou2, S. Haley3, F. Peairs4, D. Mornhinweg5, M. Bohssini6, F. Ogbonnaya6, J.Lage7, V. Tolmay8, J. Malinga9, O. Edwards10, M. Christopher11, AM. Castro12, H. Kuchel13, B. Jacobs14, I.Barclay15, J. Sheppard111 WA State Ag. Biotec. Centre, Murdoch Uni., Murdoch, WA 6150 Australia, 2CSIRO European Lab., 34980Montferrier sur Lez, France, 3Soil and Crop Sci. Dept., Colorado State Uni., Fort Collins, CO 80523-1170 USA,4 Bioag. Sci. and Pest Mgmt. Dept., Colorado State Uni., Fort Collins, CO 80523-1177 USA, 5USDA, ARS, SPA,1301 N. Western, Stillwater, OK 74075 USA, 6ICARDA, Aleppo, Syrian Arab Repub., 7CIMMYT, Apdo 370,Mexico 6, D.F. Mexico, 8ARC-Small Grain Ins., Bethlehem 9700, The Repub. of South Africa 9KARI-Njoro,NJORO, Kenya, 10CSIRO Entomology, Floreat Park, WA 6014 Australia, 11Leslie Research Centre,Toowoomba, Qld 4350 Australia, 12Dept., of Plant Sci., Faculty of Ag. Sci., Uni., of La Plata, CC31, 1900-LaPlata, Argentina, 13Australian Grain Tech., Roseworthy SA 5371 Australia, 14LongReach, Marleston, SA 5033Australia, 15Department of Ag. and Food, South Perth, WA 6151 Australia, 16 Presenting author; email:M.Cakir@murdoch.edu.auThe Russian Wheat Aphid (RWA) (Diuraphis noxia) is one of the major insect species that can cause economicyield losses to wheat (Triticum aestivum) and barley (Hordeum vulgare) growers around the world. This aphid,although is not yet present in Australia, is extremely damaging with up to 70-100% yield loses in wheatproducing lands in various parts of the world, causing significant financial losses to the grains farmers.The objectives of the study are to: 1) Identify RWA resistant wheat germplasm from around the world andcharacterize against available RWA biotypes in various countries, 2) identify molecular markers closely linkedto new resistance genes, and 3) introgress RWA resistance into more adapted wheat backgrounds from Australia,Kenya, where possible including Ug99 resistance into introgressions.We have evaluated a wide range of wheat lines from various parts of the world against a number of RWAbiotypes from Mexico, Hungary, South Africa, France and USA. Lines with moderate to good levels ofresistance were identified. To date, with the use of F2 and DH wheat populations we have mapped genes from atleast four new sources for RWA resistance on chromosomes 1DS and 7D, and closely linked SSR and DArTmarkers have been identified. Introgression of RWA resistance genes to adapted wheat and barley lines have 228
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京progressed to BC2 generation. As we have identified some wheat lines that are resistant to both RWA and Ug99in Kenya, four-way crosses were established to introgress resistance genes to both pests. Progress ofintrogressing Ug99 resistance along with RWA resistance in Kenya will further be discussed.Genotypic and phenotypic diversity analysis of a set of international wheat germplasm against RussianWheat AphidM. Cakir1,12, H. Phan1, J. Vitou2, S. Haley3, F. Peairs4, AM Castro5, R. Lindeque6, M. Bohssini7, J. Lage8, J.Malinga9, V. Tolmay6, F. Ogbonnaya7, O. Edwards10, John Sheppard111 WA State Ag. Biotec. Centre, Murdoch Uni., Murdoch, WA 6150 Australia, 2CSIRO European Lab., 34980Montferrier sur Lez, France, 3Soil and Crop Sci. Dept., Colorado State Uni., Fort Collins, CO 80523-1170 USA,4 Bioag. Sci. and Pest Mgmt. Dept., Colorado State Uni., Fort Collins, CO 80523-1177 USA, 5Dept., of PlantScience, Faculty of Agricultural Sci., Uni., of La Plata, CC31, 1900-La Plata, Argentina, 6ARC-Small GrainInstitute, Private Bag X29, Bethlehem 9700, The Republic of South Africa, 7ICARDA, Aleppo, Syrian ArabRepublic, 8CIMMYT, Apdo 370, Mexico 6, D.F. Mexico, 9KARI-Njoro, P.O. Box Private Bag- 20107, NJORO, 10 11Kenya, CSIRO Entomology, Floreat Park, WA 6014 Australia, Leslie Research Centre, Toowoomba Qld4350 Australia12 Presenting author; email: M.Cakir@murdoch.edu.auThe Russian wheat aphid (RWA), Diuraphis noxia, is one of the most damaging insect pests of wheatthroughout the World. This aphid, although is not yet present in Australia, is extremely damaging with up to70-100% yield loses in wheat producing lands in various parts of the world, causing significant financial lossesto the grains farmers.The objectives of the study are to: 1) estimate the phenotypic and genotypic diversity in the available RWAwheat germplasm from around the world against available RWA biotypes in various countries, 2) identify newresistance sources and predict the location of new resistance genes.Over 70 wheat lines were evaluated in standard seedling screening tests against a number of RWA biotypescollected from Mexico, Hungary, South Africa and France in Montpellier, France, and endemic biotypes in USA,Mexico, Kenya, South Africa, Syria and Argentina. Lines with moderate to good levels of resistance wereidentified across many biotypes. Whole genome analysis with Diversity array technology (DArT) markers hasrevealed significant genetic diversity. Results from an association analysis will also be presented. 229
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京“Genomics for Agricultural Innovation”– Japan’s national research project for utilization of genomics ininnovative crop improvementMakoto Takano (Program Officer, Agriculture, Forestry and Fisheries Research Council Secretariat, MAFF,Tokyo, JAPAN 100-8950, mtakano@affrc.go.jp)The world population is estimated to reach 9 billion in the middle of the 21st century and climate change shouldwidely affect food production that must lead to severe food shortage. Therefore, global efforts for breeding ofcereal crops with higher yield or resistance to a hostile environment are crucial in maintaining a stable worldfood supply.In the last 20 years, the extensive Rice Genome Research Program led by the National Institute ofAgrobiological Sciences (NIAS) has provided a foundation for understanding the structure and function of therice genome. More importantly, it has paved the way for the successful sequencing of the entire rice genome in2004 by an international sequencing consortium led by Japan. Subsequently, many agronomically importantgenes associated with such as high yield and resistance to both biotic and abiotic stresses were identified. The “Genomics for Agricultural Innovation” project was launched in 2008 by the Ministry of Agriculture,Forestry and Fisheries (MAFF) in order to utilize the fruits of extensive rice genome analysis for agriculturalpurposes. It serves as a core program for improving cereal crops through the advancement of basic and appliedbiotechnologies. To enrich our understanding of many agronomically important traits, wide range of researchesbased on the genomic information of rice and other cereals are being undertaken such as finding useful genes,identifying gene functions, controlling gene expression, and introducing novel genes into widely cultivatedvarieties. The project consists of 16 individual research programs from basic research such as bioinformaticsand production of research resources, to development of novel crop varieties using transgenic approach ormarker assisted selection. These programs are organically integrated in order to provide role models foreffective application of genomics in innovative crop improvement. With this project, we hope to make asignificant contribution in the worldwide efforts to develop novel cereal crops that are expected to solve theproblems associated with food, environment and energy.……………………………………………………………………………………….. 230
  • Proceedings of the 3rd International Conference of Plant 第三届植物分子育种国际学术会议摘要Molecular Breeding, Sept 5-9, 2010, Beijing, China 2010 年 9 月 5-9 日,中国,北京 农业部主管 中国农业科学院主办 全国农业核心期刊 农业科技通讯 刊号:ISSN1000-6400 CN11-2395/S 邮发代号:2-602 月刊 每月 17 日出版 单价:8 元 全年:96 元 全国各地邮局及本刊编辑部均可订阅 展示优良品种 荟萃科技成果 聚合实用技术 本刊及时报道农业最新研究成果,尤其是种业界的新产品、新技术。侧重大田,兼顾园艺,是 种植业者首选刊物。 主要栏目:人物风采、工作研究、专题论述、试 验研究、粮食作物、经济作物、蔬菜、果树、西甜 瓜、林木花卉、良种荟萃、市场信息等。内容丰富 翔实、信息量大、技术实用。 地址:100081 北京中关村南大街 12 号《农业科技通讯》编辑部 电话:010-82109664 82109665 82106276 传真:010-82109664 E-mail::tongxuna@yahoo.com.cn…………………………………………………………………………….. 231