A rice waxy mutant (M6) was generated from the japonica rice cultivar Kitaake through gamma irradiation. M6 had milky opaque kernels and lower seed size than the wild type. Sequence analysis found a 23 bp duplication in the GBSSI gene of M6, generating a stop codon and no GBSSI protein. Starch isolated from M6 lacked amylose but had similar amylopectin structure as wild type. M6 starch exhibited altered properties including higher crystallinity and different thermal and pasting properties compared to wild type starch.
impact of the mixed consortium of indigenous arbuscular mycorrhizal fungiIJEAB
The effect of inoculation of indigenousarbuscular mycorrhizal fungi (AMF) co-inoculated with Azospirillumlipoferum(strain Az204) and phosphobacteria (Bacillus megaterium - strain PB2) on the growth and yield of rice under the System of Rice Intensification (SRI) in the nursery and field was studied by conducting a field trial at Agricultural College & Research Institute, Madurai. The indigenous AMF was isolated from rice fields of this Institute and were identified as Glomus sp., Gigasporasp. and Acaulospora sp. These AMF were mass multiplied in maize plants using vermiculite as substrate and used as mixed consortium AMF. The mat nursery was prepared and AMF inoculated at the rate of 100g/m2. Also treatment was done using Azospirillum and phosphobacteria on treatment wise. At the time of transplanting seedling dip was done for the 8-day old rice seedlings using the same microbial inoculants. In the main field seed also application of mixed consortium AMF along with Azospirillum and phosphobacteria was carried out based on the treatment schedule. The results of the field trial revealed that the seedlings in the nursery showed vigorous growth and AMF colonization and spore count were recorded the maximum in the treatment with AMF, Azospirillumand 75% RDF of N and P. In the main field also there was increased growth and yield of rice plant in the same treatment due to the inoculation of mixed consortium AMF co-inoculated with Azospirillum on rice variety, ADT43 in the presence of 75% N and P. The yield of rice in this treatment recorded 11.8% higher than with 100% NP alone, besides saving 25% NP. We conclude that the mixed consortium ofindigenousAMF inoculation at the nursery and main field under SRI increased growth and grain yield of rice.
Breeding strategies for nutritional quality in major cereal cropsHeresh Puren
The presentation describes about the nutritional deficiency symptoms, deficiency status at both national and global scenario which signifies the need for breeding strategies for nutritional improvement as well as the various strategies for improvement of nutritional quality in major cereal crops.
impact of the mixed consortium of indigenous arbuscular mycorrhizal fungiIJEAB
The effect of inoculation of indigenousarbuscular mycorrhizal fungi (AMF) co-inoculated with Azospirillumlipoferum(strain Az204) and phosphobacteria (Bacillus megaterium - strain PB2) on the growth and yield of rice under the System of Rice Intensification (SRI) in the nursery and field was studied by conducting a field trial at Agricultural College & Research Institute, Madurai. The indigenous AMF was isolated from rice fields of this Institute and were identified as Glomus sp., Gigasporasp. and Acaulospora sp. These AMF were mass multiplied in maize plants using vermiculite as substrate and used as mixed consortium AMF. The mat nursery was prepared and AMF inoculated at the rate of 100g/m2. Also treatment was done using Azospirillum and phosphobacteria on treatment wise. At the time of transplanting seedling dip was done for the 8-day old rice seedlings using the same microbial inoculants. In the main field seed also application of mixed consortium AMF along with Azospirillum and phosphobacteria was carried out based on the treatment schedule. The results of the field trial revealed that the seedlings in the nursery showed vigorous growth and AMF colonization and spore count were recorded the maximum in the treatment with AMF, Azospirillumand 75% RDF of N and P. In the main field also there was increased growth and yield of rice plant in the same treatment due to the inoculation of mixed consortium AMF co-inoculated with Azospirillum on rice variety, ADT43 in the presence of 75% N and P. The yield of rice in this treatment recorded 11.8% higher than with 100% NP alone, besides saving 25% NP. We conclude that the mixed consortium ofindigenousAMF inoculation at the nursery and main field under SRI increased growth and grain yield of rice.
Breeding strategies for nutritional quality in major cereal cropsHeresh Puren
The presentation describes about the nutritional deficiency symptoms, deficiency status at both national and global scenario which signifies the need for breeding strategies for nutritional improvement as well as the various strategies for improvement of nutritional quality in major cereal crops.
Bio fortification for Enhanced Nutrition in Rice by Conventional and Molecula...Sathisha TN
Micronutrient malnutrition is widespread, especially in poor populations across the globe where daily caloric intake is confined mainly to staple cereals. Rice, which is a staple food for over half of the world's population, is low in bioavailable micronutrients required for the daily diet. Improvements of the plant-based diets are therefore critical and of high economic value in order to achieve a healthy nutrition of a large segment of the human population. Rice grain biofortification has emerged as a strategic priority for alleviation of micronutrient malnutrition
Successful colonization of roots and Plant growth promotion of sorghum (Sorgh...Premier Publishers
Pseudomonas putida (P29) and Azotobacter chroococcum (Azb19) are the efficient promising strains selected from in vitro plant growth promoting studies. These two strains were tested for their ability to promote growth of sorghum and colonize sorghum roots. Seed bacterization with P29 and Azb19 resulted in increased plant height, shoot height, root volume, leaf area and total plant dry mass. Further, bacterial inoculation also significantly increased macro-and micro-nutrient uptake by sorghum plants. Using electroporation method, pure cultures of P29 and Azb19 were transformed with pHC 60 plasmid containing gfp gene. Transformants detected by colony PCR were used to study the colonization pattern on roots of sorghum. Confocal fluorescence scanning microscope (CLSM) was used to locate the inoculants on or inside roots. Root colonization in sorghum by P29 was internal whereas Azb19 was detected on root surface. GFP-tagged Pseudomonas was predominantly detected at the root differentiation zone. In case of Azb19 small aggregates of micro-colonies were observed on the surface of the roots. The efficient sorghum root colonization by these inoculants clearly demonstrated that the introduced strains could successfully inhabit the rhizosphere and thus resulting in increased nutrient uptake. Inoculation with P29 resulted in increased uptake of P (288.5%), K (179.1%), Fe (242.7%), and Zn (168.1%) as compared to Azb19 where the uptake of P, K, Fe, Mn, and Zn increased by 142.6%, 161.6%, 199.5%, and 121.9%, respectively. On the other hand, inoculation with Azb19 could enhance better uptake of N (163.6%) as compared to P29 (133.3%). The strains also differed in their mode of root colonization.
Marker-Assisted Introgression of opaque2 and crtRB1 for Enhancement of Amino Acids and Provitamin-A in Sweet Corn.Marker-Assisted Introgression of opaque2 and crtRB1 for Enhancement of Amino Acids and Provitamin-A in Sweet Corn
Scientific opportunities and challenges of bio-fortificationGlo_PAN
Presentation by Andrew Westby, Director, Natural Resources Institute (University of Greenwich) at the launch event of the Global Panel's Biofortification Policy Brief.
Held at the All Party Parliamentary Group All-Party Parliamentary Group on Agriculture and Food for Development on 2 February 2015
Bio fortification through Genetic EngineeringBalaji Rathod
Crop Bio-fortification is the idea of breeding crops to increase their nutritional value.
Bio-fortification differs from ordinary fortification because it focuses on making plant foods more nutritious as the plants are growing, rather than having nutrients added to the foods when they are being processed.
This is an improvement on ordinary fortification when it comes to providing nutrients for the rural poor, who rarely have access to commercially fortified foods.
A description of the history, variation in methods/ approaches for biofortifying rice, benefits and challenges faced with biofortified rice and consequences for future generations..
Variability and Association of Yield and Yield Component Traits of Durum Whea...Premier Publishers
Durum wheat (Triticum turgidum L. var. durum) is a member of the Poaceae family. It is tetraploid (genomes of AABB) with 28 chromosomes (2n = 4x = 28). The use of poor yielder local durum wheat genotypes was the major critical problem of durum wheat production in Ethiopia. The present study was specifically, to evaluate genetic variations of Ethiopian advanced durum wheat lines; to investigate the important yield component traits of durum wheat; to determine the magnitude of association between yield and yield component of traits and to identify promising candidate genotypes to be used in future durum wheat breeding programmes. The study was carried out on 81 genotypes and the experiment was laid out in a triple lattice design with total of 243 experimental units. The analysis of variance showed highly significant variation among the genotypes for all traits. Most traits, have showed higher PCV and GCV values (>10%) except days to maturity. Broad sense heritability (H^2) was ranged from 57.9% to 89.7%. Future research on durum wheat crop should give emphasis on quality parameter specially required standard for pasta and Makoroni requirement of the industry demand.:
Bio fortification for Enhanced Nutrition in Rice by Conventional and Molecula...Sathisha TN
Micronutrient malnutrition is widespread, especially in poor populations across the globe where daily caloric intake is confined mainly to staple cereals. Rice, which is a staple food for over half of the world's population, is low in bioavailable micronutrients required for the daily diet. Improvements of the plant-based diets are therefore critical and of high economic value in order to achieve a healthy nutrition of a large segment of the human population. Rice grain biofortification has emerged as a strategic priority for alleviation of micronutrient malnutrition
Successful colonization of roots and Plant growth promotion of sorghum (Sorgh...Premier Publishers
Pseudomonas putida (P29) and Azotobacter chroococcum (Azb19) are the efficient promising strains selected from in vitro plant growth promoting studies. These two strains were tested for their ability to promote growth of sorghum and colonize sorghum roots. Seed bacterization with P29 and Azb19 resulted in increased plant height, shoot height, root volume, leaf area and total plant dry mass. Further, bacterial inoculation also significantly increased macro-and micro-nutrient uptake by sorghum plants. Using electroporation method, pure cultures of P29 and Azb19 were transformed with pHC 60 plasmid containing gfp gene. Transformants detected by colony PCR were used to study the colonization pattern on roots of sorghum. Confocal fluorescence scanning microscope (CLSM) was used to locate the inoculants on or inside roots. Root colonization in sorghum by P29 was internal whereas Azb19 was detected on root surface. GFP-tagged Pseudomonas was predominantly detected at the root differentiation zone. In case of Azb19 small aggregates of micro-colonies were observed on the surface of the roots. The efficient sorghum root colonization by these inoculants clearly demonstrated that the introduced strains could successfully inhabit the rhizosphere and thus resulting in increased nutrient uptake. Inoculation with P29 resulted in increased uptake of P (288.5%), K (179.1%), Fe (242.7%), and Zn (168.1%) as compared to Azb19 where the uptake of P, K, Fe, Mn, and Zn increased by 142.6%, 161.6%, 199.5%, and 121.9%, respectively. On the other hand, inoculation with Azb19 could enhance better uptake of N (163.6%) as compared to P29 (133.3%). The strains also differed in their mode of root colonization.
Marker-Assisted Introgression of opaque2 and crtRB1 for Enhancement of Amino Acids and Provitamin-A in Sweet Corn.Marker-Assisted Introgression of opaque2 and crtRB1 for Enhancement of Amino Acids and Provitamin-A in Sweet Corn
Scientific opportunities and challenges of bio-fortificationGlo_PAN
Presentation by Andrew Westby, Director, Natural Resources Institute (University of Greenwich) at the launch event of the Global Panel's Biofortification Policy Brief.
Held at the All Party Parliamentary Group All-Party Parliamentary Group on Agriculture and Food for Development on 2 February 2015
Bio fortification through Genetic EngineeringBalaji Rathod
Crop Bio-fortification is the idea of breeding crops to increase their nutritional value.
Bio-fortification differs from ordinary fortification because it focuses on making plant foods more nutritious as the plants are growing, rather than having nutrients added to the foods when they are being processed.
This is an improvement on ordinary fortification when it comes to providing nutrients for the rural poor, who rarely have access to commercially fortified foods.
A description of the history, variation in methods/ approaches for biofortifying rice, benefits and challenges faced with biofortified rice and consequences for future generations..
Variability and Association of Yield and Yield Component Traits of Durum Whea...Premier Publishers
Durum wheat (Triticum turgidum L. var. durum) is a member of the Poaceae family. It is tetraploid (genomes of AABB) with 28 chromosomes (2n = 4x = 28). The use of poor yielder local durum wheat genotypes was the major critical problem of durum wheat production in Ethiopia. The present study was specifically, to evaluate genetic variations of Ethiopian advanced durum wheat lines; to investigate the important yield component traits of durum wheat; to determine the magnitude of association between yield and yield component of traits and to identify promising candidate genotypes to be used in future durum wheat breeding programmes. The study was carried out on 81 genotypes and the experiment was laid out in a triple lattice design with total of 243 experimental units. The analysis of variance showed highly significant variation among the genotypes for all traits. Most traits, have showed higher PCV and GCV values (>10%) except days to maturity. Broad sense heritability (H^2) was ranged from 57.9% to 89.7%. Future research on durum wheat crop should give emphasis on quality parameter specially required standard for pasta and Makoroni requirement of the industry demand.:
Cereal endosperm with inhibition of starch branching enzyme (SBE) increases resistant starch content and has health benefit. For plants, endosperm starch is degraded to provide energy for seedling growth. However, whether the inhibition of SBE influences in situ degradation of starch during seedling growth is seldom reported. In this study, a normal japonica rice cultivar Wu-xiang 9915 (WX) and its derived transgenic rice line (WTR) with inhibition of SBE were cultivated in the dark only in deionized H2O. The plant growth and starch property changes were investigated during seedling development. Compared with WX, WTR showed a significantly slow plant growth. The slow degradation of starch in seed restrained the plant growth. For WX, the amylopectin and amylose were simultaneously degraded, leading to that the endosperm residual starches had similar crystalline and short-range ordered structure during seedling development. However, for WTR, the amylopectin had higher resistance to in situ degradation than amylose, and endosperm residual starches changed from CA- to CB-type and its ordered structure increased during seedling development.
Screening Of the Association of Aspergillusfungi – Macerase and Cellulase Enz...IJERA Editor
In nature, the destruction of plant wastes takes place by not a single microorganism, butby complex of microorganisms which belong to different species and genera with complex of enzymes. Using of highly active monocultures to createfungal association which produce multifunctional enzymeswith desired properties is a promising approach to create relevant and effective crop production microorganisms with beneficial properties. Among15 industrial micro my cetes the most active association was from Aspergillusniger 355 and Aspergillusawamori 1-8 which had highmacerase and cellulase enzymes activity.
Recent Milestones Achieved in Rice Genomes: Hurdles and Future Strategies by ...CrimsonpublishersMCDA
Rice is an important crop among the other cereals and considered as the model crop for function genomic studies. The rice genome size is very small 389m. The simplicity protocol of genetic transformation, physical and molecular map also developed. The recent advancement in genome sequencing and genome editing technologies has enabled us to demonstrate the potential and function of various genes for rice improvement. This spotlight presents the comprehensive overview the modern tools and resources for advance in rice genome to develop elite rice genotype which have potential tolerance against multi stresses. However, we argue the next step of rice functional genome improvement, draft genome refinement and resequencing of rice broad diversity panel genome with highly efficient technology and multidisciplinary integrated approaches to inferring gene function and future rice improvement program.
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Study on Characterization of Various Biofilms Prepared by Starch Isolated fro...ijtsrd
In the present study, the rhizome of Maranta arundinacea L., Arrowroot, was selected for a rich source of starch for the preparation of biofilm. Firstly, some physicochemical properties of the selected sample were determined by AOAC method. Furthermore, the elemental analysis of the selected sample was carried out by Energy Dispersive X ray Fluorescence EDXRF spectroscopy. Moreover, antimicrobial activities of various solvent extracts were examined by Agar well diffusion method on six tested organisms. And then, the qualitative determination of starch tests such as Iodine test and Tannic acid test were done. In addition, starch from Arrowroot powder was isolated and confirmed by FT IR spectrum. Finally, starch biofilms were prepared by using isolated starch and various ratios of plasticizers PVA, PEG, and Sorbitol. The characterizations of seven kinds of prepared biofilms were measured. Aye Mon Thida Nyo | Arnt Win | Baby San Chit Su | Mar Pi Myint | Phyu Phyu Khaing "Study on Characterization of Various Biofilms Prepared by Starch Isolated from Maranta Arundinacea L." Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-3 | Issue-5 , August 2019, URL: https://www.ijtsrd.com/papers/ijtsrd26588.pdfPaper URL: https://www.ijtsrd.com/chemistry/other/26588/study-on-characterization-of-various-biofilms-prepared-by-starch-isolated-from-maranta-arundinacea-l/aye-mon-thida-nyo
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High-value pleiotropic genes for developing multiple stress-tolerant biofort...PABOLU TEJASREE
Modern agriculture confronts multifaceted challenges, encompassing biotic and abiotic stresses alongside malnutrition. Biofortified crops emerge as a pivotal solution, augmenting nutritional quality during plant growth. By harnessing specific genes with pleiotropic effects for stress tolerance, these crops exhibit heightened yields, resilience against pests and diseases, and adaptability to environmental stressors. This innovation not only secures food safety and nutrition but also fosters the development of "high-value farms," ensuring sustainable escalation in global food productivity and stable food prices.
Conclusion: Integrating diverse transgenes and gene editing with omics approaches enhances stress tolerance and nutritional content in biofortified crops. This holistic strategy enables precise modifications to crop genomes and comprehensive insights into stress responses and nutrient metabolism, ensuring sustainable food production and nutrition security.
Microbiological and Physicochemical Properties of Wholegrain Millet Sourdough...Premier Publishers
This study enumerates the bacterial and fungal isolates responsible for the fermentation of millet sourdoughs, evaluates the antimicrobial safety of the sourdough starters and the quality of bread produced thereafter. The individual flours were spontaneously fermented in the ratio 1:1 (w/v) for 48 h. At the end of the fermentation, the microbial consortium of the sourdough meals obtained were determined and also screened for antagonistic activity against selected pathogens. All the sourdough starters revealed strong clearance zones against the selected food borne pathogens with values ranging from 5.00 to 16.1 mm. The proximate compositions of the breads were improved by the sourdough fermentation process compared with the raw flour. The moisture content of flour determined ranged from 7.48 to 11.20% while the protein, fat, ash, crude fibre and carbohydrate were 9.53 to 12.4%; 2.13 to 2.78%; 0.82 to 2.87%; 2.10 to 4.67% and 71.83 to 76.33% respectively. The carbohydrate content of the breads reduced with fermentation and ranged from 63.10% to 73.33% while the protein, ash, crude fibre and fat content increased and ranged from 11.57 to 15.31%; 2.39 to 3.08%; 1.76 to 3.62% and 7.15 to 12.32% respectively. The highest value for calcium 48.86 mg/100 g was exhibited by pearl millet bread. Sensory evaluation showed significant difference (P < 0.05) in the appearance, taste, colour, aroma and texture of the bread samples. The findings have suggested that indigenous wholegrain cereals could be employed for bread production, which has the potential to improve the nutritional profile and safety of cereal fermented foods which forms the bulk of the diet of rural dwellers in sub-Saharan Africa.
TEST CROSS PERFORMANCE AND COMBINING ABILITY OF QUALITY PROTEIN MAIZE (ZEA MA...Premier Publishers
The present study was conducted to assess the performance of test cross hybrids and estimate the combining ability of highland maize inbred lines for grain yield and yield-related traits. 40 crosses generated by crossing twenty lines with two testers and two genetic checks were evaluated using alpha lattice design with two replications at Ambo and Kulumsa agricultural research centers in 2019 main cropping season. Analyses of variances showed significant mean squares due to crosses for all studied traits that indicated genetic variation among the materials. GCA mean square due to lines showed significant differences for all traits. Similarly, GCA mean square due to testers was significant for all traits except anthesis-silking interval, while SCA mean squares were significant only for grain yield and number of ears per plant. Inbred lines viz., L7, L13, L5, L2 and L18 were good general combiners for yield and yield attributing characters. Tester CML159 was high combiner for grain yield than CML144. Among the hybrids, L5 x T1, L18 x T1 and L12 x T1 exhibited high mean values over checks and highest SCA effects for yield and yield attributing traits, thus could be used for further use in the breeding and cultivar development process.
Among the biotic stresses that affect cowpea (Vigna unguiculata L) productivity, infestation by cowpea bruchid beetles (Callosobruchus maculatus) is a major problem, causing yield losses of up to 100 %. To alleviate this problem, use of resistant varieties is a feasible approach for small-scale farmers. In Zambia, there are no reported sources of resistance to C. maculatus. The objectives of this study were: i) to evaluate certain cowpea mutants, generated at the University of Zambia, for resistance to C. maculatus; ii) to cluster the tested genotypes based on height, number of pods per plant, 100-seed weight, yield ha-1, number of eggs laid and adult emergence; and iii) to evaluate the candidate mutants for protein content. Experiments were conducted at three locations in 2014/15. The mutants, LT 11-5-2-2, BB 7-9-7-5 and BB-14-16-22, were found to be resistant to C. maculatus across locations and they out-performed their respective parents (p< 0.05) in the degree of resistance to C. maculatus. Cluster analysis, using selected agronomic parameters, showed that the selected resistant mutants (cluster B) were more similar to each other (95 %) than to Namuseba and Msandile (included as susceptible genotypes), which clustered at a similarity level of 78 % (cluster A). The mutants, LT 11-5-2-2, BB 7-9-7-5 and BB-14-16-22, showed resistance to C. maculatus, but their protein content was similar to their parents, indicating that this crucial trait had been maintained in the mutants.
www.ijera.com 1|P a g e Production of microbial enzymes by new method of cult...IJERA Editor
We have developed efficient methods for long-term culturing and selection of highly active versions of the original cultures of micromycetes – producers of enzymes. We theoretically substantiated and experimentally confirmed an advantage of growing micromycetes in a new filament-spongy immobilized growth structure on the substrate relative to the traditional method of deep cultivation of free cells in the form of pellets. When comparing a traditional with our innovative method of cultivation, many advantages of the latter are revealed, above all being the possibility of the formation of new highly selective cultures in the long process of their growth with modified culturally - morphological properties.
Similar to Characterization and Starch Properties of a Waxy Mutant in Japonica Rice Kitaake (20)
When breeding diploid potatoes, tetraploid progeny can result from the union of 2n eggs and 2n pollen in 2x-2x crosses. Thirty-three crosses were made to examine tetraploid progeny frequency in 2x-2x crosses. All crosses were between S. tuberosum dihaploids and diploid self-compatible donors, M6 and DRH S6-10-4P17. Using chloroplast counting for ploidy determination, the frequency of tetraploid progeny was as high as 45% in one of the 33 crosses. Based upon single nucleotide polymorphism (SNP) genotyping, the tetraploid progeny were attributed to bilateral sexual polyploidization (BSP), which is caused by the union of 2n egg and 2n pollen. Dihaploids were identified that produce lower frequencies of 2n eggs. The results of this study suggest that S. tuberosum dihaploids with a high frequency of 2n eggs should be avoided in 2x - 2x crosses for diploid breeding programs.
This study developed a low cost and affordable to small-scale farmers’ indirect air-cooling combined with evaporative cooling (IAC+EC) system for storage of fruit and vegetables under both arid and hot; and humid and hot climatic conditions. Field heat from freshly harvested produce should be immediately removed through cooling to the desired storage temperature. The aim of this study was to determine the effectiveness of IAC+EC system in terms of the cooling time requirement of the fresh tomato fruit. A fresh tomato cooling experiment to remove field heat during the summer month of September in Pietermaritzburg was conducted for 36 hours where the IAC+EC system was compared to storage under ambient conditions. The results showed that 16 hours was required to reduce the flesh temperature of tomatoes to 16.5°C while the flesh temperature for tomatoes under ambient conditions followed the ambient temperature profile with time of storage. The IAC+EC system reduced and maintained the microenvironment air temperature inside the coolers to 16.5°C - 19°C. The ambient temperature varied between 21 and 32°C. The results in this study are evidence that IAC+ EC system can be a choice for farmers, for cooling the fresh by reducing the field temperature after harvest.
Dioscorea rotundata is a staple food crop for millions of people in the tropical and subtropical regions. In vitro germplasm conservation is a very useful tool in yam improvement strategies but very little is known about the genetic integrity and stability of in-vitro conserved yam plants. In this study, 42 accessions from in vitro and field populations were genotyped using 11 microsatellite markers and 23 morphological descriptors to assess variability within and between accessions. Out of the 23 morphological variables used, 13 were identified as most discriminate and were used to cluster the accessions into 4 clusters using the unweighted pair group arithmetic mean average (UPGMA). Accession maintained in field as well as in in-vitro showed high genetic similarity (R2 = 0.91, p-value: 1e-04). Out of the 42 accessions analyzed, nine accessions maintained in the field and in-vitro displayed different genetic profiles. This study provided basic information on the possible somaclonal variation of yam accessions maintained through in-vitro. Further study with advanced tools such as next-generation sequencing is required to elucidate the nature of the observed variation within clones.
A study was carried out on plant density at the experimental field of the Institute of Agricultural Research for Development (IRAD) Nkolbisson, Yaoundé to determine the appropriate spacing to improve rainfed rice production in the bimodal rainfall forest zone of Cameroon. The experiment was conducted during the main cropping seasons of 2017 and 2018. The planting spacing used were 15cm x 15cm, 20cm x 20cm, 25cm x 25cm and 30cm x 30cm giving the plant populations of 444444, 250000, 160,000 and 111,111 plants / ha respectively using two varieties (Nerica 3 and Nerica 8). The experiment was laid out in a randomized complete block design with three replications. Significant differences were observed in the growth and yield across the years. Treatments were highly significant concerning the number of days to the appearance of the first flower, the number of days to 50% flowering, and the number of days to 50 % maturity. Plants were taller with more tillers and gave higher yields in 2017 than those of 2018. The spacing significantly affected the plant height, number of tillers, and panicle length for both varieties. The interaction of spacing and variety was significant for the number of tillers per m2 and the number of seeds per panicle, however, it was not for the weight of 1000 grains and the percentage of full bales. The yield components determining yield increase were the number of panicles / m2 and the number of seeds/panicles. Nerica 3 variety gave higher yields compared to the Nerica 8, the closer the spacing, the higher the yield. There were a strong significance and positive correlation between yield, number of panicles, and the number of grain per panicle. The spacing that gives the highest number of panicle per m2 was 15 cm X 15 cm and this spacing gave good yield in the region where the study was carried out.
Intermolecular interaction is the material basis for cells to achieve their functions, and protein-protein interaction is an important approach to illuminate the regulation network of biological molecules and has important theoretical significance and potential application value for revealing the activity law of life in nature. This paper mainly summarizes and analyzes the new advances and applications of modern biotechnologies in the study of protein-protein interactions, including local surface plasmon resonance (LSPR), yeast two-hybrid, GST-Pull-down, bimolecular fluorescence complementation, and coimmunoprecipitation. At the same time, the principles of different research methods for protein-protein interaction and their other applications in the field of life sciences are also discussed, all of these will provide a reference value for the analysis of protein-protein interaction and the molecular regulation mechanism of biomacromolecules.
Field experiment involving five improved onion (Allium cepa L.) varieties and carried out at Kulumsa Agricultural Research Center during the 2018 to 2019 cropping seasons under irrigated condition to identify the best performing variety for seed production to the target areas of Arsi Zone. The onion varieties included in the field experiment were (Nafis, Robaf, Nasic Red, Bombe Red, and Adama Red). The experimental design was a randomized complete block design (RCBD) with three replications. Phenological and growth parameters, seed yield, and yield components were studied. The result showed that Variety had a significant effect on most phonological and growth attributes as well as on yield attributes. The number of days to flowering and vigorously were significantly affected by variety; while plant height, days to boolting and branch number was not significantly influenced by variety. Flower stalks height and diameter, number of umbel per plant, and umbel diameter also significantly affected by variety. Seed yield per umbel and mean seed yield per hectare also showed a significance difference among varieties. The highest seed yield per hectare (1415.89 kg/ha) was recorded from Adama Red and followed by Nafis variety, whereas Nasic rerd, Bombe red and Robaf show no significance difference among each other, Therefore, it can be concluded that use of the improved onion varieties such as Adama red and Nafis is advisable and could be appropriate for onion seed production in the test area even though further testing is required to put the recommendation on a strong basis.
Rubber trees are among the important cultivated crops in Malaysia, and have contributed to the country’s overall economic growth since the 1950s. However, the existing anatomical and morphological studies are relatively insufficient. Currently, Hevea brasiliensis has been cultivated and planted commonly as a commercial planting clone, while Hevea camargoana remains to be a non-cultivated and underutilized rubber species. For many years, there only exists little information both in private plantations and government agencies that have carried out anatomical and morphological assessments on these underutilized species. There is little information about the characteristics of H.camargoana, thus raising the issue among plant breeders on how to best use this underutilized rubber species. This study attempts to investigate the taxonomic values and characteristics of Hevea brasiliensis and Hevea camargoana through anatomical and morphological studies.
Use of distributed electricity generation systems is currently increasing due to their economic and environmental benefits. Agricultural greenhouses require heat and electricity for covering their energy needs while their annual energy requirements vary significantly. Aim of the current work is the investigation of applying various distributed electricity generation systems in greenhouses. A review of different distributed generation systems currently used in various sectors as well as in greenhouses has been implemented. Various technologies are examined utilizing either renewable energies or fossil fuels in very efficient energy systems. Most of them are mature and cost-effective having lower environmental impacts compared with traditional centralized electricity generation technologies. Their use in greenhouses results in many benefits including the creation of an additional income for the farmer, reduction of carbon emissions into the atmosphere and increasing stability of the electric grid. It is suggested that distributed electricity generation systems should be used more in greenhouses when the necessary conditions are favorable.
Grain mold, considered the most important disease of sorghum, is associated with several fungal genera. The disease reduces both yield and quality. In this study, over 300 sorghum seed samples collected from Texas, Florida, and Georgia were evaluated for grain mold severity, seed weight, germination rate, and seed fungal community. Grain mold severity of the seed samples, except for those collected from Cameron, Texas, were rated 3 or higher, indicating that these sorghum lines were moderately susceptible under naturally-infected field conditions during the 2016 and 2017 growing seasons. Seed weight across surveyed locations ranged from 1.1 g to 4.0g for samples collected in Texas during the same period. Percent germination rates for samples collected in Texas ranged from 59.6% to 86.7%. Sorghum samples collected from Florida and Georgia exhibited moderately susceptible response to grain mold infection. Mean seed weight was 1.9 g for samples collected from Florida, while in Georgia, mean seed weight was 2.3 g. Germination rate was low for samples collected from Florida and Georgia. Mycological analysis of sorghum seed samples collected from farmers’ fields in Central and South Texas during the 2016 and 2017 growing seasons showed Alternaria species as the most frequently isolated fungal genus, accounting for 40% and 42 % in 2016 and 2017, followed by Fusarium incarnatum, F. acuminatum, F. equiseti, & F. semitectum Complex. In Florida and Georgia, Fusarium incarnatum, F. acuminatum, F. equiseti, & F. semitectum Complex was the most frequently recovered fungal species, accounting for 77% and 72% of the total. genera/species isolated from seed samples. Other fungal species, including Curvularia lunata, Bipolaris sp., Colletotrichum sublineola, F. verticillioides, Penicillium sp., Aspergillus flavus, F. thapsinum, F. oxysporum, F. sporotrichioides, F. graminearum, F. proliferatum, and Aspergillus niger were also isolated from sorghum seeds in various frequencies. In conclusion, the presence of large number of fungal genera associated with grain deterioration and their effect on other traits, makes management of this disease complex challenging. To identify grain mold resistant sources in a region, using the most dominant species in that region to screen the sorghum germplasm is recommended.
Potato is the second most important food crop in Kenya after maize. It is a valuable cash and food crop mainly grown by small-scale farmers. Potato is cultivated mainly under rainfed production conditions at altitudes between 1500 and 3000 metre above sea level (masl). Consequently, National Performance Trials (NPTs) test sites are located in these highlands. Recently, potato production has spread to the mid-highlands (1200-1500masl) mostly due to high food demand occasioned by population increase, and migration of small scale farmers from the densely populated highlands to the more spacious midlands. Consequently, there is increased need to breed potato varieties that can grow profitably in the warmer mid-altitudes. This development called for identification of new conventional NPT test sites located in the mid-altitudes. Six mid-altitude sites were identified and client-managed conventional NPTs carried out. Twenty six potato genotypes were evaluated for two seasons. First season crop was planted in 2016 while the second season crop was planted in 2017. There was a significant (p≤ 0.001) effect of genotypes, environments and genotype x environment interaction in the first season. Trans-Nzoia was the highest yielding site. All genotypes yielded between 10 and 25 ton/ha. In the second season, only environments showed significant (p≤ 0.001) effects. Again, Trans-Nzoia was the highest yielding site. Across the two seasons, Trans-Nzoia gave an average yield of 26.96 ton/ha followed by Kabete at 18. 21ton/ha. All the other four sites yielded less than 10 ton/ha. The high yields at Trans-Nzoia and Kabete could be due to higher altitudes in these two sites compared to the others. In order to develop potato varieties that can produce profitably in the warm mid-altitudes, breeding and early generation selections activities should be based in these areas.
The tests for the evaluation of seedling establishment, palatability and acceptability were carried out at the Teaching and Research Farm of the Department of Agricultural Technology and the Food Laboratory of the Department of Home and Rural Economics, Adamawa State College of Agriculture, Ganye, Adamawa State. The experimental designs used were Randomized Complete Block Design and the Completely Randomized Design. Some of the parameters measured were percentage seedling establishment, palatability, and acceptability tests. The highest mean percentage seedling establishment was observed in 12.00g levels of H. suaveolens whole powder treated shelled groundnut seeds (97.67, 97.00, and 94.00) at the three periods of storage. Percentage seedling establishment decreased with a decrease in levels of H. suaveolens whole powder. One of the major constraints to the use of plant materials as an alternative to synthetic insecticides is the issue of standardization in dosage of application. This work has suggested that, if groundnut seeds are to be used for sowing, they should be stored with a 12.00g level of H. suaveolens whole powder for every 300.00g for three months. However, if they are to be used for consumption, they could be stored with a 6.00g level of H. suaveolens whole powder for every 300.00g for at most three months. There was no significant difference between treated and untreated shelled groundnut seeds at the three periods of storage on the acceptability and palatability score of cooked groundnut at all dosages of application. All treatments were within the acceptable score rates even though the two rates of the synthetic insecticides, actellic dust were at the lowest acceptable score rate. The finding also showed that acceptability and palatability decreased with an increase in dosage of application. The periods of storage also had a significant impact on the mean percentage seedling establishment. It was observed that there was a higher percentage of seedling establishment when shelled groundnut seeds were stored from November- January. At this period of the year, the temperature and humidity are usually low and this might have necessitated the high state of inactivity and low performance of the bruchids compared to other periods of storage within the year.
Irrigation experiments of cocoa tend to concentrate on yield of matured cocoa trees compared to field establishment of young seedlings. Seedling survival leading to optimum population density are fundamental to obtaining maximum yield of crops. The aim of this experiment was to determine the effect of mulching and irrigation on survival of hybrid cocoa clone raised in three different growing media during the establishment phase. The experimental design was a 2 x 2 x 3 factorial arranged in a split-split plot design, with irrigation as the main plot factor, mulching as the subplot factor and growing media as the sub-sub plot factor with three replications. Cocoa pod husk (CPH) was used as the mulching material and each plant received 5kg at a rate of 5.6 t/ha. Irrigation was done daily by applying 4L of water except when it rains. Data was collected on soil moisture, plant height, leaf number, stem girth and plant survival. Results indicated that irrigation and mulching significantly (P<0.01) enhanced soil moisture. Cocoa seedlings raised in topsoil, mulched and irrigated significantly (P<0.05) increased survival rate (94.5%) compared to seedlings raised in soil without irrigation and no mulching (47.1%). Similarly, the survival rate of seedlings raised in growing media M2, mulched and irrigated (93.0%) was significantly (P<0.05) higher than similar seedlings without irrigation (73.4%). However, the survival rate of seedlings raised in M3 without mulching but irrigated was significantly (<0.05) higher 92.1% compared to seedlings raised in M3 mulched and irrigated 67.1%. Irrigation is very important in ensuring high survival rate during early field establishment. Mulching with cocoa pod husk without irrigation did not improve cocoa seedling survival.
Few years ago National Centre for Agricultural Mechanization (NCAM) launched melon sheller to reduce burden of obtaining melon oil and cake, and to enhance rural women’s productivity. This study looked at the productivity of promoted melon shelling technology and preference by rural women in Nigeria. Data were gathered from the NCAM workshop and survey of melon processors in Niger State. In the workshop, it was found that time allowed for soaking melon and covering with piece of clothes contributed to the productivity of the sheller. Results from survey revealed that 95.1% of the respondents’ preferred Internal Combustion technology and 90.2% liked electric-driven melon shellers over the manual one. Also, 69.5% obtained the shellers from local fabricators rather than NCAM. The technology led to increased turnout of melon (378kg/hr) and reduced wastages to 1.26kg out of 75.6kg. The study concluded that the promoted melon sheller is efficient and have positive implication on the rural women’s productivity and revenue. The study recommends that the rural women should put the melon sheller into its maximum capacity use. The NCAM should extend training to the local fabricators so that they can improve on the technology most especially in the areas of winnowing and washing of melon kernels.
Keeping in view of lack of recommended rates of N and NPS fertilizers, a field experiment was conducted to evaluate the effect of the newly introduced NPS fertilizer and nitrogen on growth, physiology and above ground biomass of garlic. Four NPS (0-0-0, 78.75-69-12.75, 105-92-17 and 131.25-115-21.25 kg N-P-S ha-1) and three nitrogen fertilizer rates (114.13, 228.26 and 278.33 kg N ha-1) were laid out in Randomized Complete Block Design with three replications. Significantly highest plant height (28.02 cm), leaf diameter (1.27 cm), dry and fresh weight (4.71 g and 6.11 g) and leaf length were recorded on garlic plants supplied with 105-92-17 kg N-P-S ha-1 and also the highest plant height (27.75 cm), leaf length (24.02 cm), fresh and dry weight (6.23 g and 5.04 g) were recorded on garlic plants supplied with 278.33 kg N ha-1. The interaction effect also show a significant effect in almost all the growth parameters; the early day to 50% emergence was recorded from a plot which received 228.26 kg N ha-1 and 105-92-17 kg NPS ha-1 and the highest plant height, leaf length, fresh and dry above ground biomass and leaf diameter were 29.62 cm, 25.60 cm, 6.93 g, 5.59 g and 1.4 cm, respectively were observed by the interaction of 278.33 kg N ha-1 and 105-92-17 kg N-P-S ha-1 with no significant difference with 228.26 N and 78.75-69-12.75 kg N-P-S ha−1. From this one season experiment, fertilizer rates 307.01-69-12.75 kg N-P-S ha−1 could be recommended for garlic production.
In the past, conservation effort has primarily focused on establishment of forest reserves for timber production, with minimal or no consideration to how these natural resources will help sustain or improve livelihoods of adjoining communities. This study examined the contribution of Aramoko forest reserve to the livelihoods of the adjoining communities in Ekiti State. One hundred and twenty pre-tested questionnaires were administered among the products collectors in the study area. Data collected were analysed and the result showed that 78.3 % of the respondents are married with 52.3 % of them male, while 50.8 % of the respondents aged between 51-70 years with 64.1 % having at least secondary education. Twenty-five (25) different products were collected by the respondents in the study area. These products include among others; timber with highest frequency of (120) followed by herbal plants (115), Archantina marginata (105), Thaumatococcus danielli (89), Bridelia ferruginea (70) and Irvingia garbonensis (65). The use of motor cycle accounted for 41.4 % of the respondents means of transportation, while 31.3 % of the respondents sell their product in bit at the village market. Picking, digging, plucking, cutting and uprooting were methods used for harvesting by the respondents while illegal felling (24 %) and indiscriminate bush burning (22.4 %) were the major conservation problem in the study area. Specifically, some of the products harvested are used for food, medicine, fuelwood, herbs and cultural purposes. The study recommends that research should be conducted into how these products can be artificially raised in the nursery for plantation establishment.
Cocoa farmer faces increasing challenging environment through exposure to risks factors which have impacted negatively on their production or output. Since farmers are primary producers and often times lack capacity to control risks factor, it is important to manage this factors. This study examines the cocoa farmers’ risk preferences and crop insurance perception and isolates the drivers of decision to uptake crop insurance among cocoa farmers in Ekiti state. The sample for the study consist of 200 cocoa farmers who were household heads selected through multi-stage sampling across four Local government areas of Ekiti State who are predominantly cocoa producers. Data collected on socio-demographic characteristics, perception of crop insurance and risk preferences, were analyzed using descriptive statistics and binary logistic model. Results showed that majority of the cocoa farmers were without crop insurance, with higher enrolment in Membership of cooperative society, owned their land, larger household size and lower educational level but higher level of farm experience when compared to cocoa farmers who are holding crop insurance. Farmers risk preferences showed no significant difference between farmers with or without insurance. Education (β=0.59), Household size (β=0.0029) and Debt use (β=0.02), Membership of cooperative (β=-4.53), Farming Experience (β=-2.51), Owned Land (β=-2.19) and Non-Farm Income (β=-0.65) were among the significant determinants of insurance uptake. Risk mitigating measures such as provision of necessary incentives such as improved varieties of cocoa seedling, as well as provision of fertiliser and approved pesticides, financial assistance, and simple processing technologies that produce standard cocoa bean plus a re-jigged Nigeria Agricultural Insurance Corporation (NAIC) for an improved discharge of its function are recommended.
Mitigation of climate change requires the decrease of greenhouse gas emissions into the atmosphere and the increasing use of renewable energies replacing fossil fuels. Agricultural greenhouses are energy-intensive agricultural systems using mainly fossil fuels. The use of renewable energies during their operation is limited so far. The possibility of using renewable energies for covering their energy needs has been investigated, focused on the Mediterranean region. Various sustainable energy technologies which are reliable, mature, cost-effective and broadly used in various applications are examined. These include solar-PV systems, low enthalpy geothermal energy, solid biomass burning, co-generation systems, high efficiency heat pumps and reuse of rejected industrial heat. Combined use of these systems in greenhouses can cover all their energy requirements in heat, cooling and electricity, reducing or zeroing their net CO2 emissions into the atmosphere due to operational energy use. It is concluded that depending on their local availability in Mediterranean countries, these benign energy technologies can assist greenhouse crop growers in the reduction of their carbon emissions, contributing in the achievement of the universal goal for climate change mitigation.
Field experiment was carried out under rain-fed conditions during the 2013 and 2014 cropping seasons at the Teaching and Research Farm of the Department of Agricultural Technology, Adamawa State College of Agriculture, Ganye, solely to develop an integrated pest management for the control of groundnut Aphid (Aphis craccivora Koch) [Homoptera: Aphididae]. The experimental Design used was the Randomized Complete Block Design (RCBD). Parameters measured were, average number of leaves per plot at 6 weeks after sowing, number of branches at 3 weeks after sowing, number of branches at 6 weeks after sowing, number of pods per plot, weight of harvested seeds per plot. The result obtained shows that, there was significant difference among the treatments in all the parameters measured at 0.05 level of probability using the Least Significant Difference (LSD). According to the results recorded, the highest mean yield of groundnut was obtained on plots treated with the combination of chemical and physical control methods (1444g) followed by plots treated with physical, chemical and cultural control methods combined (1296g). The highest mean number of pods per plant was recorded in the combination of physical, chemical and cultural control methods (18.00) followed by chemical and physical control methods as combined (15.00). The work shows that, the cultivation of groundnut with the control of groundnut Aphid (Aphis craccivora Koch) using integrated pest control applied as recommended facilitates better growth and guarantees good crop yield. The use of physical, chemical and cultural control method is profitable in Ganye Area of Adamawa State and is therefore suggested for use to local farmers.
Rice (Oryza sativa L.) is one of the most important food crops in the world. Purple rice, as a special rice with important nutritional and health functions, has important scientific significance and application value for genetic improvement of purple rice and breeding of new varieties through in-depth study on its agronomic and related quality traits and products processed, and analysis of its genetic basis. In this paper, the new progress of purple rice in agronomic traits, quality traits, environmental impacts on purple rice and genetic basis of purple rice seed coat in recent years were reviewed and analyzed, and the application prospects of purple rice processing products and purple gene in rice genetic improvement were also prospected. Thus, our results will provide important information and reference for breeding new purple rice varieties with good quality and high yield.
Groundnut is an important oil seed crop, grown throughout the tropics and sub tropics worldwide. It is one of the three economically important oilseed crops grown in Ethiopia. Groundnut is commonly produced by small scale farmers as food and cash crops in the study area. The area has potential to the production of Ground nut for food and nutrition security as well as export commodity. However, scarcity of varieties that fit to the environment is one of the major constraints of production. Therefore, this experiment was conducted to evaluate five Ground nut varieties and select early maturing varieties with considerable yield and agronomic traits. The field experiment was conducted in 2017 and 2018 at Abaya and varieties were planted in Randomized complete block design (RCBD). Data were collected on yield and important agronomic traits. The computed analysis of variance revealed significant variations among varieties for days to maturity, number of primary branches, number of pegs per plants, Grain yield and shelling percentage. The pooled over year mean of varieties indicated , Tole- 1 variety ( Check) is high yielding with mean grain yield of 4174.7kg followed by variety Sedi (3552.5kg/ha) and Babile local (3550.4kg/ha).Variety Sedi has special merit in terms of earliness and therefore recommended for moisture stress areas of Abaya and location with similar agro ecologies while Tole -1 (Standard check) is high yielding varieties and should be used in production until new varieties will be developed through selection/breeding program.
Richard's aventures in two entangled wonderlandsRichard Gill
Since the loophole-free Bell experiments of 2020 and the Nobel prizes in physics of 2022, critics of Bell's work have retreated to the fortress of super-determinism. Now, super-determinism is a derogatory word - it just means "determinism". Palmer, Hance and Hossenfelder argue that quantum mechanics and determinism are not incompatible, using a sophisticated mathematical construction based on a subtle thinning of allowed states and measurements in quantum mechanics, such that what is left appears to make Bell's argument fail, without altering the empirical predictions of quantum mechanics. I think however that it is a smoke screen, and the slogan "lost in math" comes to my mind. I will discuss some other recent disproofs of Bell's theorem using the language of causality based on causal graphs. Causal thinking is also central to law and justice. I will mention surprising connections to my work on serial killer nurse cases, in particular the Dutch case of Lucia de Berk and the current UK case of Lucy Letby.
This pdf is about the Schizophrenia.
For more details visit on YouTube; @SELF-EXPLANATORY;
https://www.youtube.com/channel/UCAiarMZDNhe1A3Rnpr_WkzA/videos
Thanks...!
Slide 1: Title Slide
Extrachromosomal Inheritance
Slide 2: Introduction to Extrachromosomal Inheritance
Definition: Extrachromosomal inheritance refers to the transmission of genetic material that is not found within the nucleus.
Key Components: Involves genes located in mitochondria, chloroplasts, and plasmids.
Slide 3: Mitochondrial Inheritance
Mitochondria: Organelles responsible for energy production.
Mitochondrial DNA (mtDNA): Circular DNA molecule found in mitochondria.
Inheritance Pattern: Maternally inherited, meaning it is passed from mothers to all their offspring.
Diseases: Examples include Leber’s hereditary optic neuropathy (LHON) and mitochondrial myopathy.
Slide 4: Chloroplast Inheritance
Chloroplasts: Organelles responsible for photosynthesis in plants.
Chloroplast DNA (cpDNA): Circular DNA molecule found in chloroplasts.
Inheritance Pattern: Often maternally inherited in most plants, but can vary in some species.
Examples: Variegation in plants, where leaf color patterns are determined by chloroplast DNA.
Slide 5: Plasmid Inheritance
Plasmids: Small, circular DNA molecules found in bacteria and some eukaryotes.
Features: Can carry antibiotic resistance genes and can be transferred between cells through processes like conjugation.
Significance: Important in biotechnology for gene cloning and genetic engineering.
Slide 6: Mechanisms of Extrachromosomal Inheritance
Non-Mendelian Patterns: Do not follow Mendel’s laws of inheritance.
Cytoplasmic Segregation: During cell division, organelles like mitochondria and chloroplasts are randomly distributed to daughter cells.
Heteroplasmy: Presence of more than one type of organellar genome within a cell, leading to variation in expression.
Slide 7: Examples of Extrachromosomal Inheritance
Four O’clock Plant (Mirabilis jalapa): Shows variegated leaves due to different cpDNA in leaf cells.
Petite Mutants in Yeast: Result from mutations in mitochondrial DNA affecting respiration.
Slide 8: Importance of Extrachromosomal Inheritance
Evolution: Provides insight into the evolution of eukaryotic cells.
Medicine: Understanding mitochondrial inheritance helps in diagnosing and treating mitochondrial diseases.
Agriculture: Chloroplast inheritance can be used in plant breeding and genetic modification.
Slide 9: Recent Research and Advances
Gene Editing: Techniques like CRISPR-Cas9 are being used to edit mitochondrial and chloroplast DNA.
Therapies: Development of mitochondrial replacement therapy (MRT) for preventing mitochondrial diseases.
Slide 10: Conclusion
Summary: Extrachromosomal inheritance involves the transmission of genetic material outside the nucleus and plays a crucial role in genetics, medicine, and biotechnology.
Future Directions: Continued research and technological advancements hold promise for new treatments and applications.
Slide 11: Questions and Discussion
Invite Audience: Open the floor for any questions or further discussion on the topic.
Observation of Io’s Resurfacing via Plume Deposition Using Ground-based Adapt...Sérgio Sacani
Since volcanic activity was first discovered on Io from Voyager images in 1979, changes
on Io’s surface have been monitored from both spacecraft and ground-based telescopes.
Here, we present the highest spatial resolution images of Io ever obtained from a groundbased telescope. These images, acquired by the SHARK-VIS instrument on the Large
Binocular Telescope, show evidence of a major resurfacing event on Io’s trailing hemisphere. When compared to the most recent spacecraft images, the SHARK-VIS images
show that a plume deposit from a powerful eruption at Pillan Patera has covered part
of the long-lived Pele plume deposit. Although this type of resurfacing event may be common on Io, few have been detected due to the rarity of spacecraft visits and the previously low spatial resolution available from Earth-based telescopes. The SHARK-VIS instrument ushers in a new era of high resolution imaging of Io’s surface using adaptive
optics at visible wavelengths.
Characterization and Starch Properties of a Waxy Mutant in Japonica Rice Kitaake
1. Journal of Agriculture and Crops
ISSN(e): 2412-6381, ISSN(p): 2413-886X
Vol. 4, Issue. 11, pp: 117-124, 2018
URL: https://arpgweb.com/journal/journal/14/aims
DOI: https://doi.org/10.32861/jac.411.117.124
Academic Research Publishing
Group
*Corresponding Author
117
Original Research Open Access
Characterization and Starch Properties of a Waxy Mutant in Japonica Rice
Kitaake
Long Zhang
Key Laboratory of Crop Genetics and Physiology of Jiangsu Province / Key Laboratory of Plant Functional Genomics of the Ministry of
Education, Yangzhou University, Yangzhou 225009, China
Co-Innovation Center for Modern Production Technology of Grain Crops of Jiangsu Province / Joint International Research Laboratory of
Agriculture & Agri-Product Safety of the Ministry of Education, Yangzhou University, Yangzhou 225009, China
Linglong Zhao
Key Laboratory of Crop Genetics and Physiology of Jiangsu Province / Key Laboratory of Plant Functional Genomics of the Ministry of
Education, Yangzhou University, Yangzhou 225009, China
Co-Innovation Center for Modern Production Technology of Grain Crops of Jiangsu Province / Joint International Research Laboratory of
Agriculture & Agri-Product Safety of the Ministry of Education, Yangzhou University, Yangzhou 225009, China
Xiurui Zhang
Key Laboratory of Crop Genetics and Physiology of Jiangsu Province / Key Laboratory of Plant Functional Genomics of the Ministry of
Education, Yangzhou University, Yangzhou 225009, China
Co-Innovation Center for Modern Production Technology of Grain Crops of Jiangsu Province / Joint International Research Laboratory of
Agriculture & Agri-Product Safety of the Ministry of Education, Yangzhou University, Yangzhou 225009, China
Lingshang Lin
Key Laboratory of Crop Genetics and Physiology of Jiangsu Province / Key Laboratory of Plant Functional Genomics of the Ministry of
Education, Yangzhou University, Yangzhou 225009, China
Co-Innovation Center for Modern Production Technology of Grain Crops of Jiangsu Province / Joint International Research Laboratory of
Agriculture & Agri-Product Safety of the Ministry of Education, Yangzhou University, Yangzhou 225009, China
Cunxu Wei*
Key Laboratory of Crop Genetics and Physiology of Jiangsu Province / Key Laboratory of Plant Functional Genomics of the Ministry of
Education, Yangzhou University, Yangzhou 225009, China
Co-Innovation Center for Modern Production Technology of Grain Crops of Jiangsu Province / Joint International Research Laboratory of
Agriculture & Agri-Product Safety of the Ministry of Education, Yangzhou University, Yangzhou 225009, China
Abstract
A rice waxy mutant M6 was generated from a japonica rice cultivar Kitaake through gamma irradiation. In this
study, we characterized the mutant and analyzed its starch properties. The M6 with milky opaque kernels had lower
seed length, width, and weight than wild type. The cavity in the center of starch granule might be responsible for
waxy appearance of M6 mature kernels. Sequence analysis of granule-bound starch synthase I (GBSSI) gene showed
that there was a 23 bp duplication inserted into the exon 2, generating one stop codon. No GBSSI protein was
detected in the endosperm of M6. The isolated starch showed similar ratio of short and long branch-chains of
amylopectin between M6 and wild type, but the M6 starch had no amylose. Both the M6 and wild type had A-type
starch, but the M6 starch exhibited higher relative crystallinity than wild type starch. Compared with wild type
starch, the M6 starch had significantly high swelling power, gelatinization enthalpy and breakdown viscosity and
low water solubility, gelatinization peak temperature, peak viscosity, hot viscosity, final viscosity and setback
viscosity. The M6 starch had significantly lower resistance to amylase hydrolysis than wild type starch.
Keywords: Rice waxy mutant; Endosperm starch; Molecular structure; Functional properties.
CC BY: Creative Commons Attribution License 4.0
1. Introduction
Rice (Oryza sativa L.) is one of the most important cereal crops and is the staple food for over half the world’s
population. Starch, the major storage carbohydrate in rice endosperm, consists of two distinct components: amylose
and amylopectin. Amylose is a mixture of lightly branched and long-chain linear molecules, whereas amylopectin is
a much larger molecule with a highly branched structure consisting of about 95% α-1,4 linkages and 4-5% α-1,6
linkages. According to the amylose content (AC), rice varieties are classified into waxy (0-5%), low AC (6-18%),
intermediate AC (19-23%), or high AC (>23%) types [1]. Rice seeds with high amylose levels are usually associated
with dry, hard, and poor glossy characteristics, which lower rice rating quality [2]. Therefore, control of the AC of
starch is a major objective in rice breeding.
Amylose in the rice endosperm is mainly synthesized by granule-bound starch synthase I (GBSSI), which is
encoded by the Waxy (Wx) gene located on chromosome 6 [3]. At least six Wx alleles have so far been identified:
Wxa
, Wxb
, Wxmq
, Wxop
, Wxhp
, and wx. In nonwaxy rice varieties, Wxa
and Wxb
have been recognized as being
distributed in the indica and japonica subspecies, respectively, and their expression levels are highly correlated with
2. Journal of Agriculture and Crops
118
AC in the endosperm [4]. In low AC or waxy rice varieties, a mutation in the Wx gene drastically reduces synthesis
of amylose. The leaky mutations in Wx alleles, such as Wxmq
from Milky Queen, Wxop
from the opaque endosperm
mutants and Wxhp
from Yunnan landraces, control the low AC trait [5-7]. Sequencing of Wx promoter and 5’
noncoding regions from 22 Bangladeshi rice cultivars shows that three of them with low to very low amylose lack
the G/T splice site mutation [8]. In addition, a wx mutation in the 25 tropical waxy rice varieties shows that exon 2
has a 23 bp duplication in the coding sequence [9].
In rice, AC is one of the key components influencing the eating and cooking quality (ECQ). AC is controlled
mainly by the expression levels of Wx, the activity of GBSSI and the binding characteristic of GBSSI with starch
granule [4, 5]. Different mutations in Wx have different effects on starch properties. A tyrosine residue at position
224 of Wx correlates with the formation of extra-long amylopectin chains in cultivars carrying Wxa
[10]. Using site-
directed mutagenesis, three amino acid substitutions of Wx transgenic rice lines show significant differences in
GBSSI activities, AC and starch physicochemical properties [11]. An amino acid substitution in Wxhp
allele reduces
the binding of GBSSI to starch granule and AC, affecting the pasting property of starch in rice seeds [5]. Therefore,
it is necessary to investigate the effects of different Wx mutations on the properties of starch.
Waxy rice, also known as glutinous rice, is widely used for food products, such as glutinous rice crackers and
glutinous rice wine in Thailand, Laos, and particularly in China. In Thailand, most of the waxy rice varieties are
photoperiod sensitive landraces [9]. Being photoperiod sensitive landraces, waxy rice varieties are difficult to be
popularized and applied in large areas. Compared with other rice varieties, the Kitaake variety has a life cycle of
only nine weeks, four times a year, and is insensitive to photoperiod changes, which can greatly accelerate the
functional genetic studies and new varieties breeding of rice [12]. In China rice market, a lot of waxy rice varieties
are cultivated by 60
Co gamma irradiation, such as Funuo 101, Funuoyou 396, Guifunuo, Yangfunuo 1 and
Yangfunuo 4. In this study, we isolated a rice waxy mutant from the japonica rice cultivar Kitaake through gamma
irradiation. The characterization and starch properties of M6 were investigated. The major objective of this study was
to reveal the molecular mechanism of mutation and the structural and functional properties of starch in order to use
the mutant effectively in waxy rice breeding program in the future.
2. Materials and Methods
2.1. Plant Materials
The waxy mutant M6 was isolated from a 60
Co-irradiated mutant pool of japonica rice cultivar Kitaake. The
Kitaake and M6 were grown in a paddy field at Yangzhou University during the natural growing season. Mature
seeds were harvested and used to isolate starch.
2.2. Scanning Electron Microscopy
Seeds were randomly selected for phenotypic analysis. To obtain cross sections, seeds were mounted on
aluminum specimen stubs with adhesive tabs, coated with gold, and observed under an environmental scanning
electron microscope (Philips XL-30) at 5 kV.
2.3. RNA Extraction and Sequence Analysis
Total RNA were extracted from seedlings of wild type and M6 using an RNAprep pure Plant Kit (TIANGEN,
Beijing). First strand complementary DNA (cDNA) was synthesized with oligo (dT18) based on a PrimeScript
Reverse Transcriptase Kit (Takara, Japan). The wild type and M6 GBSSI cDNA sequence was cloned using primers
5’-ATGTCGGCTCTCACCACGTCCCA-3’ and 5’-AGGAGAACGTGGCTGCTCCTTGA -3’, and the PCR
product was introduced into the pEASY-Blant Vector (Transgen, Beijing), and transformed into E. coli strain DH5α.
The recombinant plasmid was sequenced with an ABI Prism 3730 XL DNA Analyzer (PE Applied Biosystems,
USA).
2.4. Protein Extraction and Western Blot Analysis
Mature seed endosperms were ground to powder in liquid N2. The powder was then suspended in the extraction
buffer consisting of 50 mM Tris/HCl, pH 8.0, 0.25 M sucrose, 2 mM DTT, 2 mM EDTA, and 1 mM
phenylmethylsulphonyl fluoride. After incubation on ice for 1 h, the homogenate was centrifuged for 20 min at
14,000 g, and the supernatants were transferred to new centrifuge tubes. Proteins were resolved by SDS-PAGE and
transferred electrophoretically to polyvinylidene difluoride membrane. The antibodies used were anti-GBSSI rabbit
antibody (Immunogen, Wuhan, China) diluted 1:5000, and anti-HSP82 (Beijing Protein Innovation) diluted 1:10000,
and horseradish peroxidase-linked secondary antibody (Beyotime, Shanghai, China) diluted 1:5000.
2.5. Isolation of Starch from Brown Rice
The brown rice was soaked in distilled water at 4 °C for 24 h and extensively ground in a mortar. The ground
sample was filtered through five layers of cotton cloth and filtered with 100-, 200-, and 400-mesh sieves,
successively. The sample was centrifuged at 4000 g for 5 min and washed with distilled water. Starch was washed
three times with water and twice with anhydrous ethanol. Finally, samples were dried at 40 °C and ground through a
100-mesh sieve.
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2.6. Measurement of Starch Molecular Structure
Apparent amylose content (AAC) of starch was determined following the iodine colorimetric method described
by Wang, et al. [13]. The molecular weight distribution of starch was analyzed using an Agilent Technologies gel-
permeation chromatography (GPC) 220 system according to the method described by Lin, et al. [14].
2.7. X-Ray Diffraction (XRD) Analysis of Starch
Starch XRD patterns were obtained with an X-ray power diffractometer (D8, Bruker, Germany). All samples
were treated in a desiccator with a saturated solution of NaCl to maintain a constant humidity (relative
humidity=75%) for 7 days prior to XRD analysis. The relative crystallinity was determined as described by Wei, et
al. [15].
2.8. Swelling Power and Water Solubility Determination of Starch
Swelling power and water solubility of starch were determined according to the method of Konik-Rose, et al.
[16] with some modifications. Starch samples mixed with water (2%, w/v) was put in a 2 mL centrifuge tube and
heated in a water bath at 95 °C for 30 min with regular gentle shock. The sample was cooled to room temperature
and centrifuged at 8000 g for 20 min. The swelling power was the weight ratio of precipitated gel to dry starch.
2.9. Thermal Property Analysis of Starch
Thermal properties of starch granules were investigated with a differential scanning calorimetry (DSC) (200-F3,
NETZSCH, Germany). Three milligrams of starch was mixed with 9 μL of distilled water, and sealed in an
aluminum pan. The sample was then heated from room temperature to 130 °C at a rate of 10 °C/min.
2.10. Pasting Property Analysis of Starch
The pasting properties of starch were evaluated with a rapid visco analyzer (RVA) (RVA-3D, Newport
Scientific, Narrabeen, Australia). Two grams of starch was dispersed in 25 mL distilled water and subjected to
gelatinization analysis. A programmed heating and cooling cycle was used, where the sample was held at 50 °C for 1
min, heated to 95 °C at a rate of 12 °C/min, maintained at 95 °C for 2.5 min, cooled to 50 °C at a rate of 12 °C/min,
and then held at 50 °C for 1.4 min.
2.11. Enzyme Hydrolysis Analysis of Starch
The starch was hydrolyzed by porcine pancreatic α-amylase (PPA) and Aspergillus niger amyloglucosidase
(AAG). For PPA hydrolysis, ten milligrams of starch was suspended in 2 mL of enzyme solution (0.1 M phosphate
sodium buffer, pH 6.9, 25 mM NaCl, 5 mM CaCl2, 0.02% NaN3, 50 U PPA (Sigma A3176)). For AAG hydrolysis,
ten milligrams of starch was suspended in 2 mL of enzyme solution (0.05 M acetate buffer, pH 4.5, 5 U AAG
(Sigma A7095)). The hydrolyses of PPA and AAG were conducted in a shaking water bath with continuous shaking
(100 rpm) at 37 and 55 °C, respectively. After hydrolysis, starch slurry was quickly centrifuged (5000 g) for 5 min.
The soluble carbohydrate in the supernatant was determined to quantify the hydrolysis degree using the anthrone-
H2SO4 method.
2.12. Statistical Analysis
For sample characterization, at least three replicate measurements were performed. All data represent the means
± standard deviation (n=3). The results were analyzed using the Student’s t-test to examine differences. Results with
a corresponding probability value of p<0.01 were considered to be statistically significant.
3. Results and Discussion
3.1. Phenotypic Characterization of M6
Numerous mutants with defective endosperm were screened from the 60
Co-induced mutant library of japonica
rice cultivar Kitaake and an opaque-kernel mutant was isolated and named M6. Throughout the vegetative growth
stage, the mutant plants displayed no significant differences from the wild type plants. The mature grains of M6 were
phenotypically similar to its wild type (Fig. 1A), while the brown rice of M6 was opaque and presented a milky-
white appearance (Fig. 1B-D). Iodine staining is a sensitive and convenient method for the detection of amylose in
various tissues. When the seeds of M6 were cut transversely and stained with an iodine solution, a typical reddish
color of waxy starch was revealed in the endosperm (Fig. 1C-F;) [17]. Scanning electron microscopy analysis of
transverse sections indicated that the compound starch granules in both wild type and M6 endosperm cells were
irregularly polyhedral and densely packed, while some cavities were only observed in the center of the starch granule
in M6 (Fig. 1G, H). This phenotype in M6 starch granule was similar to the low amylose materials such as Y268F
and E410D [11]. Seed size and weight measurements showed that seed thickness was largely comparable between
the wild type and M6, but the seed length, seed width and thousand seed weight were significantly reduced in the M6
(Fig. 1I-L). These results suggested that the mutation in M6 might affect the amylose synthesis in endosperm.
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120
3.2. Molecular Analysis of M6 Mutation
It is generally accepted that amylose synthesis is carried out by GBSS. Cereals contain two forms of GBSS,
GBSSI and GBSSII, and GBSSI is responsible for amylose synthesis in storage tissues, such as endosperm [18].
Thus, the cDNA sequences of GBSSI were amplified by PCR and sequenced. Comparison of the sequences of wild
type and the M6 revealed that the M6 allele carried 23 bp insertion (red font) in the exon 2 of GBSSI (Fig. 2A). The
23 bp insertion was the duplication of the 23 bp (black underline) in front of them and generated a premature stop
codon that allowed translation of only the first 57 amino acids of the GBSSI protein (Fig. 2A, B). Thus, M6 was a
loss-of-function mutant in GBSSI. Subsequently, we used western blot to analyze GBSSI protein in rice endosperm.
As shown in Fig. 2C, the GBSSI antibody specially recognized the endogenous GBSSI protein band in wild type but
not in the M6. In summary, these results demonstrated that the mutation of GBSSI was responsible for the M6
phenotypes.
3.3. Structural Properties of M6 Starch
The AAC of wild type and M6 starch as determined by iodine colorimetry is given in Table 1. The AAC of M6
starch was 1.2%, which was significantly lower than that of wild type starch (14.3%). The molecular weight
distribution of starch as determined by GPC is shown in Fig. 3A. In general, the GPC chromatogram of isoamylase-
debranched starch exhibits three peaks. The Peaks 1 and 2 represent the short-branch chains (A and short B chains)
and long-branch chains (long B chains) of amylopectin, respectively, and the Peak 3 is amylose [19]. The M6
contained very low apparent amylose. Therefore, only two peaks (Peak 1 and 2) were detected in the GPC profile of
M6 starch. Whereas for wild type starches, three peaks were detected in its GPC profiles due to the simultaneous
existence of amylose and amylopectin (Fig. 3A). The percentage of the peak area in GPC profile can reflect the
molecular weight distribution of starch, and the area ratio of Peak 1 and Peak 2 can be used as an index of the extent
of amylopectin branching; the higher the ratio, the higher the branching degree [20]. As is shown in Table 1, M6
starch consisted of approximately 76.0% amylopectin short branch-chains and 24.0% amylopectin long branch-
chains, and had 3.2% amylopectin branching degree, while wild type starch contained 65.0% amylopectin short
branch-chains, 21.5% amylopectin long branch-chains, and 13.5% amylose, and had 3.0% amylopectin branching
degree.
The XRD patterns of starches are presented in Fig. 3B. Both wild type and M6 starches had the characteristics of
A-type crystallinity with strong reflection peaks at about 15°and 23°, and an unresolved doublet at around 17°and
18° 2θ (Fig. 3B), which was in conformity with the characteristics of normal cereal starches [21]. The relative
crystallinity of M6 starch was 33.4%, which was higher than that of wild type starch (27.1%). This result was in
accordance with that the relative crystallinity is negatively related to amylose [22].
3.4. Functional Properties of M6 Starch
The swelling power and water solubility of wild type and M6 starches at 95°C are shown in Table 1. The M6
starch had higher swelling power and lower water solubility than wild type starch. The swelling power is a measure
of the water-holding capacity of starch after being heated in water, cooled, and centrifuged, while the water
solubility reflects the degree of dissolution during the starch swelling procedure [23]. Amylose is considered to
contribute to the inhibition of water absorption and swelling of starch, whereas amylopectin tends to promote the
process [24]. The M6 starch had higher amylopectin content and lower AC than wild type starch, which might
contribute to its higher swelling power. The lower water solubility of M6 starch might be related to its lower AC,
which could not leach out of the starch granules into the water.
The thermal properties of starch samples were determined by DSC, and their thermograms and thermal
parameters are given in Fig. 4A and Table 2. Compared with wild type starch, M6 starch exhibited lower
gelatinization peak temperature and higher gelatinization enthalpy. The gelatinization peak temperature correlates
positively with AC [25]. Starch with higher amylopectin contents can easily form more crystalline structures within
granules; this type of starch also requires more energy to melt and uncoil the double helix structure during
gelatinization [26]. In the present study, the M6 starch had a lower AC and higher crystallinity, and therefore
required lower gelatinization temperature and more energy for gelatinization than wild type starch.
The pasting properties of wild type and M6 starches measured by RVA are presented in Fig. 4B, and their
pasting parameters are given in Table 2. Compared with the wild type starch, M6 starches had significantly different
pasting properties because of their different molecular structure. The M6 starch had significantly lower peak
viscosity, hot viscosity, final viscosity, setback viscosity, and peak time than wild type starch. This finding confirms
the suggestion that rice starch with a low amylose is more prone to gelatinization, in agreement with previous studies
on maize [27]. In contrast, as for the breakdown viscosity, M6 starch was significantly higher than wild type starch,
possibly because amylose intertwines with amylopectin in wild type starch, which helped to maintain the integrity of
the starch granules [28].
The time courses of PPA and AAG hydrolysis of starches are presented in Fig. 4C and Fig. 4D. A biphasic
hydrolysis trend by PPA or AAG was observed in wild type and M6 starches with an initial rapid hydrolysis of the
amorphous region followed by a decreased hydrolysis, which was in agreement with previous studies [13]. The
hydrolysis rate of M6 starch by PPA or AAG was markedly higher than wild type starch. PPA hydrolyzes starch
begins firstly from granule surface, and then it penetrates into granule interior and degrades starch from inside to
outside, while AAG hydrolyses starch from the outer surface of the granule [29]. Susceptibility of starch to PPA and
AAG attack is influenced by factors such as AC, amylose to amylopectin ratio, crystalline structure, granule
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integrity, porosity of granules, and structural inhomogeneities [30]. In the present study, the low AC and cavity
structure in M6 starch led to that M6 starch was enzymatically hydrolyzed faster than the wild type starch.
4. Conclusion
In conclusion, the M6 was a loss-of-function mutant of GBSSI, which produced a waxy endosperm composed of
amylose-free starch granules. Because of the differences in molecular structure between M6 and wild type, M6 starch
contained higher relative crystallinity, swelling power, gelatinization enthalpy, and breakdown viscosity, but lower
water solubility, gelatinization peak temperature, peak viscosity, hot viscosity, final viscosity, setback viscosity, and
resistance to PPA and AAG hydrolysis than wild type starch. These findings could provide some practical
information on the potential usefulness of the waxy mutant.
Acknowledgements
This study was financially supported by grants from the Natural Science Foundation of Jiangsu Province
(BK20160461), the China Postdoctoral Science Foundation (2018T110561), the Qing Lan Project of Jiangsu
Province, the Talent Project of Yangzhou University, and the
Priority Academic Program Development of Jiangsu Higher Education Institutions.
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Table-1. Apparent Amylose Contents (AAC), Molecular Weight Distributions, Swelling Powers (SP), and Water Solubilities (WS) of Wild Type
and M6 Starches
AAC (%)
Molecular Weight Distribution
SP (g/g) WS (%)
ASB (%) ALB (%) AM (%) ASB/ALB
WT 14.3±0.3 65.0±1.3 21.5±0.1 13.5±1.2 3.0±0.1 17.2±0.1 5.9±0.3
M6 1.2±0.2** 76.0±0.5** 24.0±0.5** ND 3.2±0.1 35.5±0.4** 3.2±0.3**
ASB, amylopectin short branch-chains; ALB, amylopectin long branch-chains; AM, amylose; ND, not detected.
Data are shown as means ± standard deviation (n=3) and compared with wild type by Student’s t-test (**P<0.01).
Table-2. The Parameters of Thermal and Pasting Properties of Wild Type and M6 Starches
Thermal Parameter Pasting Parameter
To (°C) Tp (°C) Tc (°C) ΔT (°C) ΔH (J/g) PV (mPa s) HV (mPa s) BV (mPa s) FV (mPa s) SV (mPa s) PT (min)
WT 62.0±0.2 69.8±0.2 77.2±0.1 15.1±0.1 10.0±0.2 1817±1 1632±6 185±4 1946±1 314±7 6.8±0.1
M6 61.7±0.3 68.5±0.4** 77.0±1.5 15.3±1.3 11.5±0.1** 1640±5** 841±8** 799±4** 974±13** 133±21** 3.7±0.0**
To, onset temperature; Tp, peak temperature; Tc, conclusion temperature; ΔT, gelatinization temperature range (Tc-To); ΔH, gelatinization enthalpy; PV, peak
viscosity; HV, hot viscosity; BV, breakdown viscosity (PV-TV); FV, final viscosity; SV, setback viscosity (FV-HV); PT, peak time.
Data are shown as means ± standard deviation (n=3) and compared with wild type by Student’s t-test (**P<0.01).
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Fig-1. Phenotypic analyses of M6. (A, B) The morphologies of grains (A) and seeds (B) of wild type (above) and M6 (below). Scale bar = 1 mm.
(C, D) Transverse sections of representative wild type (C) and M6 (D) dry seeds. Scale bar = 1mm. (E, F) Transverse sections of representative
wild type (E) and M6 (F) dry seeds with iodine-staining. Scale bar = 1mm. (G, H) Scanning electron microscope images of transverse sections of
the wild type (G) and M6 (H) dry seeds. Red arrows indicate the cavity in the center of starch granule. Scale bar = 5 μm. (I-K) Quantification of
seed length (I), seed width (J), and seed thickness (K) of wild type and M6 (n=30). (L) 1000-seed weight of wild type and M6 (n=3). Data are
shown as means ± standard deviation, and compared with wild type by Student’s t-test (**P<0.01)
Fig-2. Molecular identification of M6 mutation. (A) Structure of the GBSSI gene and positions of mutation site. The insertion of 23 bp in the
exon 2 leads to frame shift and creates premature stop codon in M6. (B) Amino acid sequence of GBSSI in M6. (C) Western blot analysis of
GBSSI in the mature seed of the wild type and M6. Anti-HSP82 antibody is used as a loading control
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Fig-3. GPC Profiles (A) and XRD Patterns (B) of Wild Type and M6 Starches. The Relative Crystallinity is Given in Parentheses
Fig-4. DSC the Rmograms (A), Pasting Profiles (B), PPA Hydrolyses (C), and AAG Hydrolyses (D) Of Wild Type and M6 Starches