1) Zone lines were observed on the lower stems of mature soybean plants in multiple Midwestern states. Diaporthe longicolla was consistently isolated from these zone lines.
2) Molecular analysis confirmed the identity of 10 representative isolates as D. longicolla. Phylogenetic analysis placed these isolates in a well-supported clade with known D. longicolla cultures.
3) Greenhouse pathogenicity tests reisolated D. longicolla from zone lines on inoculated soybean plants, fulfilling Koch's postulates and demonstrating that D. longicolla is associated with and causes the black zone lines observed on mature soybean stems.
Inter Simple Sequence Repeats (ISSR) markers were utilized to identify the levels of heritable varieties and patterns of the populace structure among the five populaces of Pteris biaurita, a natural fern in India. A comprehensive examination was directed in three replicates at 2013-14 seasons in the Western Ghats, South India. Five wild P. biaurita, accessions (maiden hair) were assessed for genotyping studies. Results demonstrated a pivotal discrepancy among genotypes for they were characterized in view of this uniqueness in four groups by the genetic cluster examination. In this trial, ISSR primers amplified 63 polymorphic groups. In view of the genetic identity data, genotypes were figured and differed from 0.5714 to 0.6984. The percentage of polymorphism indicated predominant genotype that may be utilized for the conservation of species. ISSR appeared to be an obliging marker for prediction of genotype inside a closed group of inter specific populace in the investigation territory
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.
This document discusses genetic diversity analysis of grape (Vitis vinifera L.) germplasm in India using microsatellite markers. A total of 42 grape genotypes were analyzed using 7 microsatellite markers. A total of 45 alleles were detected among the genotypes. The microsatellites grouped the genotypes into two main clusters (A and B) based on morphological and genetic characteristics, with subclusters differentiating seeded vs seedless fruits and pigmented vs non-pigmented fruits. The study found high genetic variability among the Indian grape germplasm and that microsatellite markers are a reliable tool for diversity and breeding programs.
This study aims to quantify populations of the entomopathogenic fungus Beauveria bassiana in soil samples from kudzu patches infested with the invasive kudzu bug, Megacopta cribraria, in North Carolina. Soil samples were collected from seven kudzu patches and B. bassiana colony forming units (CFUs) were quantified over seven days. Preliminary results found low B. bassiana populations that decreased over time, with no difference between patch locations. Molecular identification using PCR and sequencing will determine pathogenic strains to evaluate for biocontrol of the kudzu bug.
1) The study analyzed genetic diversity in 66 finger millet accessions from Ethiopia and Eritrea using RAPD markers.
2) A total of 123 RAPD fragments were amplified using 15 primers, of which 89 (72%) were found to be polymorphic.
3) Genetic similarity between accessions ranged from 0.585 to 0.984. Cluster analysis grouped the 66 accessions into nine clusters at a similarity index of 0.83, showing high genetic variability among the accessions.
This document describes the identification of FLOWERING LOCUS T (FT) through an activation tagging screen in Arabidopsis thaliana. The 1733 mutant tagged line flowered early and had terminal flowers. Sequencing of the tagged gene revealed it to be FT. Overexpression of FT using the 35S promoter recapitulated the 1733 phenotype, flowering very early with few leaves. FT was found to act partially downstream of CONSTANS (CO) to promote flowering in response to day length. Unlike many floral regulators, the FT protein sequence does not suggest it directly controls transcription.
Identification and pathogenicity of fusarium and phomopsis foliar diseases of...Premier Publishers
Research on foliage disease of Jatropha curcas was conducted in Sokoto, Kebbi and Zamfara States of Nigeria to determine the occurrence, incidence and severity of the diseases. Fusarium and Phomopsis species were the fungal pathogens found to be responsible for the disease on J. curcas in the study area. A spore count of the isolates was made and used as inocula in the pathogenicity trial in glasshouse of the department to prove Kochs’ postulate. Results from the farmers’ field revealed that, highest incidence (81.00%) and severity (53.33%) of Phomopsis leaf blight was recorded in Tsaki of Sokoto State, while Janbaki in Kebbi State had the highest incidence (75.33%) and severity (60.00%) of Fusarium leaf blight. The surveys conducted showed that, J. curcas planted in lowland areas tend to be more prone to the fungal leaf blight particularly those close to water source. In the pathogenicity trial, results indicated that, there was no significant difference in the methods of inoculation and number of days after inoculation with respect to incidence and severity of leaf blight. It is recommended that fungicides that can be used for the management of fungal leaf blight of J. curcas should be identified.
Inter Simple Sequence Repeats (ISSR) markers were utilized to identify the levels of heritable varieties and patterns of the populace structure among the five populaces of Pteris biaurita, a natural fern in India. A comprehensive examination was directed in three replicates at 2013-14 seasons in the Western Ghats, South India. Five wild P. biaurita, accessions (maiden hair) were assessed for genotyping studies. Results demonstrated a pivotal discrepancy among genotypes for they were characterized in view of this uniqueness in four groups by the genetic cluster examination. In this trial, ISSR primers amplified 63 polymorphic groups. In view of the genetic identity data, genotypes were figured and differed from 0.5714 to 0.6984. The percentage of polymorphism indicated predominant genotype that may be utilized for the conservation of species. ISSR appeared to be an obliging marker for prediction of genotype inside a closed group of inter specific populace in the investigation territory
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.
This document discusses genetic diversity analysis of grape (Vitis vinifera L.) germplasm in India using microsatellite markers. A total of 42 grape genotypes were analyzed using 7 microsatellite markers. A total of 45 alleles were detected among the genotypes. The microsatellites grouped the genotypes into two main clusters (A and B) based on morphological and genetic characteristics, with subclusters differentiating seeded vs seedless fruits and pigmented vs non-pigmented fruits. The study found high genetic variability among the Indian grape germplasm and that microsatellite markers are a reliable tool for diversity and breeding programs.
This study aims to quantify populations of the entomopathogenic fungus Beauveria bassiana in soil samples from kudzu patches infested with the invasive kudzu bug, Megacopta cribraria, in North Carolina. Soil samples were collected from seven kudzu patches and B. bassiana colony forming units (CFUs) were quantified over seven days. Preliminary results found low B. bassiana populations that decreased over time, with no difference between patch locations. Molecular identification using PCR and sequencing will determine pathogenic strains to evaluate for biocontrol of the kudzu bug.
1) The study analyzed genetic diversity in 66 finger millet accessions from Ethiopia and Eritrea using RAPD markers.
2) A total of 123 RAPD fragments were amplified using 15 primers, of which 89 (72%) were found to be polymorphic.
3) Genetic similarity between accessions ranged from 0.585 to 0.984. Cluster analysis grouped the 66 accessions into nine clusters at a similarity index of 0.83, showing high genetic variability among the accessions.
This document describes the identification of FLOWERING LOCUS T (FT) through an activation tagging screen in Arabidopsis thaliana. The 1733 mutant tagged line flowered early and had terminal flowers. Sequencing of the tagged gene revealed it to be FT. Overexpression of FT using the 35S promoter recapitulated the 1733 phenotype, flowering very early with few leaves. FT was found to act partially downstream of CONSTANS (CO) to promote flowering in response to day length. Unlike many floral regulators, the FT protein sequence does not suggest it directly controls transcription.
Identification and pathogenicity of fusarium and phomopsis foliar diseases of...Premier Publishers
Research on foliage disease of Jatropha curcas was conducted in Sokoto, Kebbi and Zamfara States of Nigeria to determine the occurrence, incidence and severity of the diseases. Fusarium and Phomopsis species were the fungal pathogens found to be responsible for the disease on J. curcas in the study area. A spore count of the isolates was made and used as inocula in the pathogenicity trial in glasshouse of the department to prove Kochs’ postulate. Results from the farmers’ field revealed that, highest incidence (81.00%) and severity (53.33%) of Phomopsis leaf blight was recorded in Tsaki of Sokoto State, while Janbaki in Kebbi State had the highest incidence (75.33%) and severity (60.00%) of Fusarium leaf blight. The surveys conducted showed that, J. curcas planted in lowland areas tend to be more prone to the fungal leaf blight particularly those close to water source. In the pathogenicity trial, results indicated that, there was no significant difference in the methods of inoculation and number of days after inoculation with respect to incidence and severity of leaf blight. It is recommended that fungicides that can be used for the management of fungal leaf blight of J. curcas should be identified.
Experimental validation, genetic map saturation and gene flow pilot study in ...CIAT
The document summarizes research that aimed to: 1) validate single nucleotide polymorphism (SNP) markers in common bean, 2) saturate an existing genetic linkage map with SNPs, and 3) study the potential of SNPs to determine gene flow patterns in a wild-weedy-crop complex. 92 out of 130 SNPs were validated as true polymorphisms. 136 SNPs were added to the genetic map, covering previously unmapped regions. Analysis of SNP haplotypes in a wild-weedy-crop complex from Colombia found no clear differentiation between biological forms. Both directions of gene flow appeared possible based on haplotype frequencies.
This document describes a DNA marker-based technology developed to identify citrus rootstocks at the seedling stage. Traditionally, Rough lemon and Rangpur lime are preferred rootstocks but are difficult to distinguish from Galgal at an early stage. Galgal is an undesirable rootstock due to susceptibility to diseases. The new technology uses microsatellite markers and PCR to differentiate Rough lemon, Rangpur lime, and Galgal based on presence or absence of DNA fragments. It provides a non-destructive method to ensure nurseries are supplying quality rootstock varieties. The technique has been transferred to agricultural organizations to benefit citrus farmers.
Evaluation of genetic diversity of soybean introductions and north american a...hiensh02
This document evaluates the genetic diversity of 87 soybean plant introductions and 18 major ancestors of North American soybean germplasm using RAPD and SSR markers. Genetic distances among the 105 genotypes ranged from 0.08 to 0.76. Clustering analysis grouped the genotypes into 11 clusters of varying stability. Several groups of plant introductions were distinct from the major ancestors and may provide new genetic variation for soybean breeding programs seeking to broaden the genetic base.
Characterisation of some Ribes L. accessions from Turkey based on SSRs patternsAgriculture Journal IJOEAR
This document summarizes a study that analyzed genetic variability among 7 Ribes alpinum, 2 Ribes bieberstenii, and 1 Ribes uva-crispa accessions from Turkey using SSR (microsatellite) markers. A total of 10 SSR primers were used, producing 172 bands between 50-330 base pairs in length. 157 of these bands were polymorphic, representing 91.2% genetic diversity. The SSR patterns allowed for delineation of the Ribes accessions at both the specific and intraspecific levels, providing additional data for characterization of the Ribes gene pool in Anatolia.
This document analyzed the genetic diversity of 50 Asian bitter gourd genotypes using morphological traits and molecular markers. Key findings:
1. Significant variation was found for yield and other traits based on morphological analysis, indicating genetic diversity. The highest yielding genotype was Sel-2.
2. Molecular analysis using RAPD and ISSR markers found high levels of polymorphism, with ISSR showing more polymorphic bands.
3. Cluster analyses based on morphological, RAPD, ISSR, and combined data grouped genotypes into clusters largely correlating with geographical origin and domestication status. The analyses demonstrate large genetic variability in the collection.
An Overview of Sudden Death Syndrome in Soybean (Glycine max) and Molecular R...Chloe Siegel
This document provides an overview of sudden death syndrome in soybeans, which is caused by the fungus Fusarium virguliforme. It discusses the symptoms, yield losses, and transmission process. It also examines the proteins and toxins produced by the fungus that are responsible for damage. Additionally, it outlines the plant hormone pathways and defense mechanisms involved in the plant's response. Finally, it notes that while no highly resistant cultivars currently exist, research aims to better understand genetic resistance to develop more resistant varieties.
16. varietal characterization of tomato cultivars based on rapd markersVishwanath Koti
This study characterized 24 tomato cultivars using RAPD markers. Eleven primers produced 100 bands, of which 89.39% were polymorphic. Each cultivar had unique DNA sequences not found in others. The primers OPC-02, OPC-19, OPD-19, OPD-18 and OPC-08 generated the most unique bands, producing 13 unique bands among 10 cultivars. The combination of OPB-10 with either OPC-19 or OPB-08 was sufficient to identify all 24 tomato cultivars.
Resistance/tolerance to root-knot nematode Meloidogyne incognita (Kofoid and ...Premier Publishers
This experiment was carried out in a Randomized Complete Block Design (40 X 10m plot) to evaluate cacao clones for resistance and tolerance against Meloidogyne incognita in infested field. The ten cocoa clones evaluated are MXC67, T86/2, PA150, LCTEEN, T12/11, T53/5, T101/15, T65/7, ICS1 and AMAZ 15-15. Pre-planting survey revealed ten genera of phytonematodes, these were; Meloidogyne spp., Pratylenchus spp., Helicotylenchus spp. Paralongidorus spp., Eutylenchus spp., Scutellonema spp., Hemicyclophora spp., Xiphinema spp., Longidorus spp., Anguillulina spp., Psilenchus spp., and Tetylenchus spp. Meloidogyne spp. had the highest frequency of occurrence and highest population in 250g soil (40 and 28,234 respectively), Paralongidorus spp. was next in population. Based on gall index, nematode reproduction factor and growth parameters, MXC67, T86/2, PA150, T101/15 and T53/5 were susceptible to the nematode. Two other clones, T65/7 and ICS1 were tolerant. A high degree of resistance was exhibited by LCTEEN, T12/11 and AMAZ 15-15. Compared with F3 Amazon and Amelonado varieties, the two most famous cocoa varieties in Nigeria, four clones (LCTEEN, T65/7, ICSI and AMAZ 15-15) were superior to F3 Amazon and Amelonado. This study showed that planting of resistance/tolerant cacao clones in nematode infested soil will drastically reduce seedling failure experienced by farmers. This tolerant and resistant clones identified should be included in breeding programme for resistance.
This document lists 49 references to scientific papers related to DNA barcoding and herbal medicine. The references are listed by number and include the authors, year published, title of the paper, and journal or publication details. They cover topics like using DNA barcoding to identify plant species for quality control in herbal medicine, investigating genetic variation and chemical properties of medicinal plant populations, and evaluating DNA markers for authenticating medicinal plant species.
9 diversity of meloidogyne in martiniqueAna Karenina
The study characterized Meloidogyne species infecting banana roots across plantations in Martinique, Guadeloupe, and French Guiana. A total of 96 nematode isolates were collected and identified using esterase and malate dehydrogenase enzyme phenotypes, as well as perineal patterns. The major species found were M. arenaria at 61.9% of isolates and M. incognita at 34.3%. Intraspecific variability was detected within M. arenaria and M. incognita using the enzyme analysis. RAPD markers also revealed genetic variation within and between species, with M. arenaria showing 61.6% polymorphic variation within isolates. This high intraspecific diversity in M. aren
Biogeography and polyphasic approach of pseudomonas strains from agriculture ...Alexander Decker
This document summarizes a study on the isolation and characterization of Pseudomonas strains from agricultural land in Madhya Pradesh, India. Soil samples were collected from different districts and 50 Pseudomonas strains were isolated. The isolates were characterized using biochemical tests as well as molecular techniques including DNA isolation, PCR-RFLP of 16S rDNA, RAPD-PCR, and REP-PCR fingerprinting. Genetic diversity analysis showed the isolates had genetic similarity with some variation in their genomes. The isolates exhibited potential for use as plant growth-promoting rhizobacteria and biocontrol agents based on their biochemical profiles.
Rice stress related gene expression analysisRonHazarika
The document summarizes research on stress response proteins in rice (Oryza sativa). It identifies 17 proteins commonly up-regulated and 3 commonly down-regulated in response to drought, heat, and salinity stress. It analyzes the protein with the highest interaction for each group and identifies 10 similar proteins in each family. It examines the proteins' physicochemical properties, 3D structures, and functions in plant defense. The study finds the proteins structurally similar but functionally diverse, concluding they help rice cope with stress through complex regulatory interactions.
This document discusses allele mining as an advanced technique for crop improvement. It begins by defining allele mining as searching for different alleles located at the same locus. It then outlines several key steps and techniques for allele mining, including Eco-TILLING based allele mining, sequencing based allele mining, and association mapping based allele mining. The document provides details on each technique, including requirements, procedures, advantages, and examples of crops studied. It emphasizes that allele mining is important for unlocking genetic variation stored in germplasm collections in order to develop crops with improved traits.
Responses of wheat seedling to varying moisture conditions and relationship b...Agriculture Journal IJOEAR
— The following study was conducted to estimate the genotypic differences among 30 wheat (Triticum aestivum L.) genotypes under different moisture regimes and relationship between morphological and molecular characterization. Eight seedling parameters root length (RL), shoot length (SL), root fresh weight (RFW), shoot fresh weight (SFW), root dry weight (RDW), shoot dry weight (SDW), chlorophyll rate (CR) and survival rate (SR) were studied at four different soil moisture conditions (T 1 40%,T 2 60%,T 3 80%,T 4 100%) using two factor factorial complete randomized design (CRD). Significant differences among genotypes were observed by analysis of variance. For heritability estimates, survival rate showed lowest heritability under all the treatments. Principal components analysis accounted 81.4% variation in T 1 , 81.9% in T2, 87.7% in T3 and 84.7% in T4 conditions in first PC. Selected diverse genotypes were further fingerprinted with 10 ISSR markers. A total of 74 DNA fragments were detected and 72.7% of was polymorphic. The amplified DNA fragments were ranged from 4 (UBC-809) to 11 (UBC-808). PIC values were ranged from 0.32 to 0.81. Cluster analysis grouped the genotypes into 4 clusters on the basis of molecular and phenotypic characterization under T4 normal conditions whereas under T1 (moisture stress) conditions genotypes were grouped into 5 clusters explaining genotypic differences under different moisture conditions. The present results showed that phenotypic difference in wheat seedling expression under different water regimes is accompanied with molecular basis, which offer a prospective to enhance wheat adaptation under moisture stress conditions.
Molecular marker to identify gynoecious lines in bitter gourdSwati Saxena
This document discusses the identification of molecular markers associated with the gynoecious trait in bitter gourd. Twenty-four gynoecious plants were screened using 200 RAPD and 28 ISSR markers. One ISSR primer amplified a 1000 base pair fragment present in all gynoecious plants but absent in two monoecious varieties. This fragment was repeatably amplified and could serve as a diagnostic marker for gynoecy, allowing identification of the trait at an early stage for hybrid seed production.
The document summarizes an experiment conducted by Jennifer Griffith, Taylor Wadley, and Daniel Crall to study the effects of light intensity and pH on the regeneration of African violets (Saintpaulia ionantha) through tissue culture. They tested five different pH levels of growth media (4.0-8.0) and three lights varying in color temperature (4100K, 6500K, 4100K) and color rendering index (89, 84, 70). They hypothesized the greatest shoot production would occur with a pH of 6.0 and light with a 4100K color temperature and 89 CRI, as these conditions are closest to established protocols and light spectra promoting growth.
This document describes the development of 160 novel simple sequence repeat (SSR) markers in bitter gourd (Momordica charantia L.) through enriched genomic libraries. Genomic DNA from bitter gourd was used to construct libraries enriched for 10 different repeat motifs. Of the 3,072 clones screened, 93.7% contained microsatellite repeats. Unique primer pairs were designed and validated for 151 loci. Genetic diversity analysis of 51 loci among 54 accessions found 20% were polymorphic. The markers distinguished 15 Indian varieties and 78.4% were transferable across six Momordica species. The new SSR markers will be useful for genetic studies in bitter gourd.
Genetic variability, heritability, genetic advance, genetic advance as percen...Premier Publishers
This document summarizes a study on the genetic variability, heritability, genetic advance, and character associations of 49 Ethiopian mustard landraces. The study found significant genetic variability among the accessions for all traits measured. Traits like seed yield, oil yield, and plant height showed high genotypic and phenotypic variation, indicating potential for selection. Heritability was highest for thousand seed weight, days to flowering, stand percent, and oil quality traits. Positive correlations were found between seed yield and traits like oil content, oil yield, plant height and seed yield per plant. Primary branches and oil yield showed direct positive effects on seed yield per plot. Seed yield, oil content, oil yield and primary branches were determined to be
This document describes a study that developed a new method called amplified functional DNA restriction analysis (AFDRA) to analyze the diversity of catechol 2,3-dioxygenase (C23O) genes in soil bacteria. C23O genes code for enzymes important for degrading aromatic pollutants. The researchers used AFDRA to analyze C23O genes from reference strains and soil isolates. They found that AFDRA generated distinct restriction patterns that clustered the isolates into four groups, consistent with sequence analysis. AFDRA also allowed them to determine the predominant C23O gene variants present in environmental DNA extracts from soil samples. The study demonstrates that AFDRA provides a rapid way to assess functional gene diversity in cultures and
Derrick Guidry is seeking a leadership position utilizing over 16 years of experience in team building, leadership mentoring, project management, and process improvement. He has a proven track record of reducing costs through resource management proposals and implementing changes to schedules. Currently a Team Leader at Kelsey Seybold Clinic, he is responsible for motivating teams to meet objectives and has developed a process improvement plan to reduce turnover, costs, and increase productivity. Derrick holds a Bachelor's degree in Operations Management from the University of Houston and has over 16 years of experience leading teams in logistics, healthcare, and wireless industries.
Experimental validation, genetic map saturation and gene flow pilot study in ...CIAT
The document summarizes research that aimed to: 1) validate single nucleotide polymorphism (SNP) markers in common bean, 2) saturate an existing genetic linkage map with SNPs, and 3) study the potential of SNPs to determine gene flow patterns in a wild-weedy-crop complex. 92 out of 130 SNPs were validated as true polymorphisms. 136 SNPs were added to the genetic map, covering previously unmapped regions. Analysis of SNP haplotypes in a wild-weedy-crop complex from Colombia found no clear differentiation between biological forms. Both directions of gene flow appeared possible based on haplotype frequencies.
This document describes a DNA marker-based technology developed to identify citrus rootstocks at the seedling stage. Traditionally, Rough lemon and Rangpur lime are preferred rootstocks but are difficult to distinguish from Galgal at an early stage. Galgal is an undesirable rootstock due to susceptibility to diseases. The new technology uses microsatellite markers and PCR to differentiate Rough lemon, Rangpur lime, and Galgal based on presence or absence of DNA fragments. It provides a non-destructive method to ensure nurseries are supplying quality rootstock varieties. The technique has been transferred to agricultural organizations to benefit citrus farmers.
Evaluation of genetic diversity of soybean introductions and north american a...hiensh02
This document evaluates the genetic diversity of 87 soybean plant introductions and 18 major ancestors of North American soybean germplasm using RAPD and SSR markers. Genetic distances among the 105 genotypes ranged from 0.08 to 0.76. Clustering analysis grouped the genotypes into 11 clusters of varying stability. Several groups of plant introductions were distinct from the major ancestors and may provide new genetic variation for soybean breeding programs seeking to broaden the genetic base.
Characterisation of some Ribes L. accessions from Turkey based on SSRs patternsAgriculture Journal IJOEAR
This document summarizes a study that analyzed genetic variability among 7 Ribes alpinum, 2 Ribes bieberstenii, and 1 Ribes uva-crispa accessions from Turkey using SSR (microsatellite) markers. A total of 10 SSR primers were used, producing 172 bands between 50-330 base pairs in length. 157 of these bands were polymorphic, representing 91.2% genetic diversity. The SSR patterns allowed for delineation of the Ribes accessions at both the specific and intraspecific levels, providing additional data for characterization of the Ribes gene pool in Anatolia.
This document analyzed the genetic diversity of 50 Asian bitter gourd genotypes using morphological traits and molecular markers. Key findings:
1. Significant variation was found for yield and other traits based on morphological analysis, indicating genetic diversity. The highest yielding genotype was Sel-2.
2. Molecular analysis using RAPD and ISSR markers found high levels of polymorphism, with ISSR showing more polymorphic bands.
3. Cluster analyses based on morphological, RAPD, ISSR, and combined data grouped genotypes into clusters largely correlating with geographical origin and domestication status. The analyses demonstrate large genetic variability in the collection.
An Overview of Sudden Death Syndrome in Soybean (Glycine max) and Molecular R...Chloe Siegel
This document provides an overview of sudden death syndrome in soybeans, which is caused by the fungus Fusarium virguliforme. It discusses the symptoms, yield losses, and transmission process. It also examines the proteins and toxins produced by the fungus that are responsible for damage. Additionally, it outlines the plant hormone pathways and defense mechanisms involved in the plant's response. Finally, it notes that while no highly resistant cultivars currently exist, research aims to better understand genetic resistance to develop more resistant varieties.
16. varietal characterization of tomato cultivars based on rapd markersVishwanath Koti
This study characterized 24 tomato cultivars using RAPD markers. Eleven primers produced 100 bands, of which 89.39% were polymorphic. Each cultivar had unique DNA sequences not found in others. The primers OPC-02, OPC-19, OPD-19, OPD-18 and OPC-08 generated the most unique bands, producing 13 unique bands among 10 cultivars. The combination of OPB-10 with either OPC-19 or OPB-08 was sufficient to identify all 24 tomato cultivars.
Resistance/tolerance to root-knot nematode Meloidogyne incognita (Kofoid and ...Premier Publishers
This experiment was carried out in a Randomized Complete Block Design (40 X 10m plot) to evaluate cacao clones for resistance and tolerance against Meloidogyne incognita in infested field. The ten cocoa clones evaluated are MXC67, T86/2, PA150, LCTEEN, T12/11, T53/5, T101/15, T65/7, ICS1 and AMAZ 15-15. Pre-planting survey revealed ten genera of phytonematodes, these were; Meloidogyne spp., Pratylenchus spp., Helicotylenchus spp. Paralongidorus spp., Eutylenchus spp., Scutellonema spp., Hemicyclophora spp., Xiphinema spp., Longidorus spp., Anguillulina spp., Psilenchus spp., and Tetylenchus spp. Meloidogyne spp. had the highest frequency of occurrence and highest population in 250g soil (40 and 28,234 respectively), Paralongidorus spp. was next in population. Based on gall index, nematode reproduction factor and growth parameters, MXC67, T86/2, PA150, T101/15 and T53/5 were susceptible to the nematode. Two other clones, T65/7 and ICS1 were tolerant. A high degree of resistance was exhibited by LCTEEN, T12/11 and AMAZ 15-15. Compared with F3 Amazon and Amelonado varieties, the two most famous cocoa varieties in Nigeria, four clones (LCTEEN, T65/7, ICSI and AMAZ 15-15) were superior to F3 Amazon and Amelonado. This study showed that planting of resistance/tolerant cacao clones in nematode infested soil will drastically reduce seedling failure experienced by farmers. This tolerant and resistant clones identified should be included in breeding programme for resistance.
This document lists 49 references to scientific papers related to DNA barcoding and herbal medicine. The references are listed by number and include the authors, year published, title of the paper, and journal or publication details. They cover topics like using DNA barcoding to identify plant species for quality control in herbal medicine, investigating genetic variation and chemical properties of medicinal plant populations, and evaluating DNA markers for authenticating medicinal plant species.
9 diversity of meloidogyne in martiniqueAna Karenina
The study characterized Meloidogyne species infecting banana roots across plantations in Martinique, Guadeloupe, and French Guiana. A total of 96 nematode isolates were collected and identified using esterase and malate dehydrogenase enzyme phenotypes, as well as perineal patterns. The major species found were M. arenaria at 61.9% of isolates and M. incognita at 34.3%. Intraspecific variability was detected within M. arenaria and M. incognita using the enzyme analysis. RAPD markers also revealed genetic variation within and between species, with M. arenaria showing 61.6% polymorphic variation within isolates. This high intraspecific diversity in M. aren
Biogeography and polyphasic approach of pseudomonas strains from agriculture ...Alexander Decker
This document summarizes a study on the isolation and characterization of Pseudomonas strains from agricultural land in Madhya Pradesh, India. Soil samples were collected from different districts and 50 Pseudomonas strains were isolated. The isolates were characterized using biochemical tests as well as molecular techniques including DNA isolation, PCR-RFLP of 16S rDNA, RAPD-PCR, and REP-PCR fingerprinting. Genetic diversity analysis showed the isolates had genetic similarity with some variation in their genomes. The isolates exhibited potential for use as plant growth-promoting rhizobacteria and biocontrol agents based on their biochemical profiles.
Rice stress related gene expression analysisRonHazarika
The document summarizes research on stress response proteins in rice (Oryza sativa). It identifies 17 proteins commonly up-regulated and 3 commonly down-regulated in response to drought, heat, and salinity stress. It analyzes the protein with the highest interaction for each group and identifies 10 similar proteins in each family. It examines the proteins' physicochemical properties, 3D structures, and functions in plant defense. The study finds the proteins structurally similar but functionally diverse, concluding they help rice cope with stress through complex regulatory interactions.
This document discusses allele mining as an advanced technique for crop improvement. It begins by defining allele mining as searching for different alleles located at the same locus. It then outlines several key steps and techniques for allele mining, including Eco-TILLING based allele mining, sequencing based allele mining, and association mapping based allele mining. The document provides details on each technique, including requirements, procedures, advantages, and examples of crops studied. It emphasizes that allele mining is important for unlocking genetic variation stored in germplasm collections in order to develop crops with improved traits.
Responses of wheat seedling to varying moisture conditions and relationship b...Agriculture Journal IJOEAR
— The following study was conducted to estimate the genotypic differences among 30 wheat (Triticum aestivum L.) genotypes under different moisture regimes and relationship between morphological and molecular characterization. Eight seedling parameters root length (RL), shoot length (SL), root fresh weight (RFW), shoot fresh weight (SFW), root dry weight (RDW), shoot dry weight (SDW), chlorophyll rate (CR) and survival rate (SR) were studied at four different soil moisture conditions (T 1 40%,T 2 60%,T 3 80%,T 4 100%) using two factor factorial complete randomized design (CRD). Significant differences among genotypes were observed by analysis of variance. For heritability estimates, survival rate showed lowest heritability under all the treatments. Principal components analysis accounted 81.4% variation in T 1 , 81.9% in T2, 87.7% in T3 and 84.7% in T4 conditions in first PC. Selected diverse genotypes were further fingerprinted with 10 ISSR markers. A total of 74 DNA fragments were detected and 72.7% of was polymorphic. The amplified DNA fragments were ranged from 4 (UBC-809) to 11 (UBC-808). PIC values were ranged from 0.32 to 0.81. Cluster analysis grouped the genotypes into 4 clusters on the basis of molecular and phenotypic characterization under T4 normal conditions whereas under T1 (moisture stress) conditions genotypes were grouped into 5 clusters explaining genotypic differences under different moisture conditions. The present results showed that phenotypic difference in wheat seedling expression under different water regimes is accompanied with molecular basis, which offer a prospective to enhance wheat adaptation under moisture stress conditions.
Molecular marker to identify gynoecious lines in bitter gourdSwati Saxena
This document discusses the identification of molecular markers associated with the gynoecious trait in bitter gourd. Twenty-four gynoecious plants were screened using 200 RAPD and 28 ISSR markers. One ISSR primer amplified a 1000 base pair fragment present in all gynoecious plants but absent in two monoecious varieties. This fragment was repeatably amplified and could serve as a diagnostic marker for gynoecy, allowing identification of the trait at an early stage for hybrid seed production.
The document summarizes an experiment conducted by Jennifer Griffith, Taylor Wadley, and Daniel Crall to study the effects of light intensity and pH on the regeneration of African violets (Saintpaulia ionantha) through tissue culture. They tested five different pH levels of growth media (4.0-8.0) and three lights varying in color temperature (4100K, 6500K, 4100K) and color rendering index (89, 84, 70). They hypothesized the greatest shoot production would occur with a pH of 6.0 and light with a 4100K color temperature and 89 CRI, as these conditions are closest to established protocols and light spectra promoting growth.
This document describes the development of 160 novel simple sequence repeat (SSR) markers in bitter gourd (Momordica charantia L.) through enriched genomic libraries. Genomic DNA from bitter gourd was used to construct libraries enriched for 10 different repeat motifs. Of the 3,072 clones screened, 93.7% contained microsatellite repeats. Unique primer pairs were designed and validated for 151 loci. Genetic diversity analysis of 51 loci among 54 accessions found 20% were polymorphic. The markers distinguished 15 Indian varieties and 78.4% were transferable across six Momordica species. The new SSR markers will be useful for genetic studies in bitter gourd.
Genetic variability, heritability, genetic advance, genetic advance as percen...Premier Publishers
This document summarizes a study on the genetic variability, heritability, genetic advance, and character associations of 49 Ethiopian mustard landraces. The study found significant genetic variability among the accessions for all traits measured. Traits like seed yield, oil yield, and plant height showed high genotypic and phenotypic variation, indicating potential for selection. Heritability was highest for thousand seed weight, days to flowering, stand percent, and oil quality traits. Positive correlations were found between seed yield and traits like oil content, oil yield, plant height and seed yield per plant. Primary branches and oil yield showed direct positive effects on seed yield per plot. Seed yield, oil content, oil yield and primary branches were determined to be
This document describes a study that developed a new method called amplified functional DNA restriction analysis (AFDRA) to analyze the diversity of catechol 2,3-dioxygenase (C23O) genes in soil bacteria. C23O genes code for enzymes important for degrading aromatic pollutants. The researchers used AFDRA to analyze C23O genes from reference strains and soil isolates. They found that AFDRA generated distinct restriction patterns that clustered the isolates into four groups, consistent with sequence analysis. AFDRA also allowed them to determine the predominant C23O gene variants present in environmental DNA extracts from soil samples. The study demonstrates that AFDRA provides a rapid way to assess functional gene diversity in cultures and
Derrick Guidry is seeking a leadership position utilizing over 16 years of experience in team building, leadership mentoring, project management, and process improvement. He has a proven track record of reducing costs through resource management proposals and implementing changes to schedules. Currently a Team Leader at Kelsey Seybold Clinic, he is responsible for motivating teams to meet objectives and has developed a process improvement plan to reduce turnover, costs, and increase productivity. Derrick holds a Bachelor's degree in Operations Management from the University of Houston and has over 16 years of experience leading teams in logistics, healthcare, and wireless industries.
El documento presenta un kit para estudiantes que incluye herramientas como horarios de estudio, descansos, atención, memoria, motivación y pasos para resolver problemas. El kit describe elementos como establecer horarios de estudio fijos, tomar descansos para renovar la energía, mantener la concentración, recopilar información de forma permanente, sentir pasión por lo que se estudia, y establecer conexiones para resolver problemas.
The document provides information on the Zika virus, including its history, epidemiology, transmission, signs and symptoms, complications, diagnosis, and current situation. It discusses how the virus was first identified in 1947 in Uganda in monkeys and humans in 1952. It outlines its spread to Africa, Asia, the Pacific islands, and the Americas. It also summarizes Brazil reporting over 500,000 suspected Zika cases and the observed increase in Guillain-Barré syndrome and microcephaly linked to the outbreak.
Fractional Fourier Transform: Fractional Wiener Filter in ScilabMarcos Gonzalez
Este documento describe el desarrollo de una aplicación en Scilab para el tratamiento de señales usando la transformación de Fourier fraccionaria. Presenta el marco teórico de la transformación de Fourier fraccionaria y sus propiedades. Luego, detalla el algoritmo propuesto para la discretización e implementación numérica de la transformación. Finalmente, describe la metodología y desarrollo de la aplicación en Scilab, incluyendo funciones como rectángulo, triángulo, coseno, seno y chirp.
SlidesA Comparison of GPU Execution Time Prediction using Machine Learning an...Marcos Gonzalez
This document compares GPU execution time prediction using machine learning techniques and analytical modeling. It begins with introductions to parallel programming models, GPU architectures, and machine learning techniques. It then describes testing methodology where algorithms were run on various NVIDIA GPUs and datasets were collected to compare machine learning approaches like linear regression and random forests to an analytical BSP-based model for GPU execution time prediction. The goal is to determine which approach more accurately predicts execution times.
The document describes three individuals - a 6-year-old girl named Kaorymosquera who has short curly black hair, dark eyes, and was wearing a yellow and white shirt; a 26-year-old elegant woman named Yulitsamosquera who has long straight black hair, brown eyes, an oval face and was wearing a blue cotton dress and sandals; and an elderly man in his early 80s who is short and thin with white hair, dark eyes, an oval face, and was wearing a blue wool jacket, black bonnet and brown pants.
Inter Simple Sequence Repeats (ISSR) markers were utilized to identify the levels of heritable varieties and patterns of the populace structure among the five populaces of Pteris biaurita, a natural fern in India. A comprehensive examination was directed in three replicates at 2013-14 seasons in the Western Ghats, South India. Five wild P. biaurita, accessions (maiden hair) were assessed for genotyping studies. Results demonstrated a pivotal discrepancy among genotypes for they were characterized in view of this uniqueness in four groups by the genetic cluster examination. In this trial, ISSR primers amplified 63 polymorphic groups. In view of the genetic identity data, genotypes were figured and differed from 0.5714 to 0.6984. The percentage of polymorphism indicated predominant genotype that may be utilized for the conservation of species. ISSR appeared to be an obliging marker for prediction of genotype inside a closed group of inter specific populace in the investigation territory.
The document summarizes a study that aimed to clarify the taxonomy of fungal pathogens causing apple ring rot in China. Researchers applied genealogical concordance phylogenetic species recognition to 24 fungal isolates from apple and pear with disease symptoms, along with reference isolates. Phylogenetic analysis of sequences from four nuclear loci revealed two distinct species - one including an isolate previously identified as Botryosphaeria berengeriana f. sp. piricola, and the other including an ex-epitype isolate of B. dothidea. While the two fungi induced different disease symptoms on apple and pear, the cryptic species showed sufficient genetic and biological differences from B. dothidea to be described as a new combination, Botryos
Growth Pattern, Molecular Identification and Bio molecules Analysis of FOMITO...journal ijrtem
Abstract : Fomitopsis feei, a brown rot fungus is identified tentatively using morphological characteristics and confirmed phylogenetically by 28S rDNA analysis and sequence was submitted in EMBL Nucleotide Sequence Database. Its growth pattern was studied on eight different solid media and found to be good on Malt extract agar medium. Biomolecules such as proteins and lipid were screened qualitatively and estimated quantitatively. Aminoacid analysis by chromatography and fatty acid analysis by FAME were also done and revealed that tryptophan (20.53%), valine (20.51%) and cis-linoleic acid (43.38%) and palmetic acid (17.88%) were in high percentage.
Key words : Fomitopsis feei, growth, molecular identification and biomolecules
The document summarizes research on micropropagation and genetic transformation of Scutellaria ocmulgee, a threatened medicinal plant. Leaf and stem explants from S. ocmulgee were used in thin cell layer culture (tTCL) on different growth media. Murashige and Skoog medium with benzylaminopurine and naphthaleneacetic acid produced the highest number of shoot buds. Shoots were rooted and hardened into plants. An Agrobacterium-mediated transformation protocol was also developed using tTCL to insert marker genes. Results showed tTCL is effective for micropropagation and provides a system for genetic transformation of S. ocmulgee.
Phylotype Analysis of Ralstonia Solanacearum Causing Bacterial wilt in Eggpla...ijtsrd
Eggplant is prone to attack by several pests including bacteria, fungi, nematodes and insects. In this study, we have analyzed phylotype of bacterial wilt Ralstonia solanacearum infection in eggplant plants collected from Bhubaneswar Orissa in India. Bacterial wilt symptomatic five plant samples were collected from brinjal field in Bhubaneswar in 2016. The samples were macerated in sterile distilled water and grown on Kelman's triphenyltetrazolium chloride TZC agar media. Total genomic DNA of the bacterium were extracted and subjected to PCR amplification using the R. solanacearum specific universal primer pair 759 760. An expected single 280 bp fragment amplified in all the samples confirmed the identity of these as Ralstonia. To reconfirmed isolate of bacterium, the amplicon was sequenced in sequencer. In NCBI blast, the nucleotide sequence was 100 similar with Ralstonia solanacearum strain RS lpxC DOB 1 AB910593 and the sequence was submitted in NCBI database under Acc. No. KY393266. To determined phylotype of strain used specific multiplex PCR with phylotype specific primers Nmult 21F1 2, Nmult 22InF, Nmult 23AF, Nmult 22RR revealed that all the five infected samples belonged to phylotype I as a 144 bp amplicon were observed in agarose gel. On the basis of above finding concluded that the bacterial wilt infected eggplant collected from Bhubaneswar was Ralostonia solanacearum, Phylotype I. Rakesh Kumar | Ramachandran, E. | Koteshwar Yadav "Phylotype Analysis of Ralstonia Solanacearum Causing Bacterial wilt in Eggplants in Orissa in India" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-3 | Issue-3 , April 2019, URL: https://www.ijtsrd.com/papers/ijtsrd21580.pdf
Genome Sequencing in Finger Millet
Genome size estimation
SOLiD Sequencing Technology
Illumina Sequencing Technology
Gene prediction and functional annotation of genes
Mining of plant transcription factors and other genes
This document describes a new species of Rhododendron found in the Gaoligong Mountains of Yunnan, China. Morphologically, Rhododendron baihuaense can be distinguished from related species by its absent scales on mature leaves, 2-5 flowers per inflorescence, funnel-shaped white corollas, and slender erect styles. Molecular analysis of DNA barcoding regions supports R. baihuaense as genetically distinct from close relatives. Both morphological and genetic evidence demonstrate this represents a new species, Rhododendron baihuaense, endemic to Yunnan.
The document discusses a study that analyzed 18 grape hybrids and 7 parents from India using 12 microsatellite markers. The goals were to confirm the hybrid nature of the progenies and construct a molecular database of the parents. Cluster analysis grouped the parents and hybrids into two major groups based on genetic relationships. Three markers (VVMD-32, VVS-29, and VVS-2) were able to confirm the hybrid nature of all progenies by detecting polymorphisms between the male and female parents that were then present in the hybrids. The study confirmed the reliability of using microsatellite markers for parentage analysis and genetic studies in grapevines.
This document summarizes a study that used molecular markers to analyze genetic transformation in grafted Citrus sinensis (sweet orange) plants. Random amplified polymorphic DNA (RAPD) and simple sequence repeat (SSR) markers were used to compare DNA from scion mother plants, grafted plants, and rootstock plants from Manipur. Three primers detected genetic differences between the samples. One RAPD primer found an amplicon present in rootstocks and grafts but not scions, indicating transmission from stock to scion. Another RAPD primer produced an excess amplicon in grafts not found in stock or scion. One SSR primer detected the absence of an amplicon in two grafts
- The study characterized 13 microsatellite markers for Calochortus gunnisonii, a montane lily species in the central and southern Rocky Mountains, using next-generation DNA sequencing.
- The markers were found to be polymorphic, with a mean of 4.97 alleles per locus. Observed and expected heterozygosity ranged from 0.077 to 0.871 and 0.213 to 0.782, respectively.
- The markers will be useful for investigating population structure, genetic diversity, and demographic history of C. gunnisonii across its range in the Rocky Mountains. They also showed potential for cross-species amplification in other Calochortus species.
- Experiments tested the effect of extracellular self-DNA (exDNA) and heterologous DNA on the growth of 6 species from different taxonomic groups, including bacteria, fungi, algae, plants, protozoa and insects.
- Treatments with conspecific exDNA produced a concentration-dependent growth inhibition in all species, whereas heterologous DNA did not cause inhibition except in one bacterial species.
- The results suggest exDNA may have a general inhibitory effect on biological systems, providing a potential mechanism for self-inhibition and negative feedback observed in different organisms. Further investigation is needed to understand the molecular mechanisms of this effect.
Rapid Impact Assessment of Climatic and Physio-graphic Changes on Flagship G...Arvinder Singh
‘NATIONAL CONFERENCE ON MAN AND ENVIRONMENT’October 15 – 16, 2012
Organized by
Department of Zoology and Environmental Sciences, Punjabi University, Patiala (Pb.) – 147 002, India
Abstracts Of Presentations At The 2015 APS Annual MeetingBecky Goins
This document contains abstracts from presentations given at the 2015 APS Annual Meeting. The abstracts describe various plant pathology research studies and are grouped alphabetically by first author's name. The summaries provided are from three of the abstracts within the document.
Abstract— An experiment stand of clonal orchard of masson pine, which included the 123 plus trees of 8 provenances collected from 8 provinces of Southern China, was founded at Jingshan County of Hubei province. Randomly amplified polymorphic DNA (RAPD) technique was applied to assess genetic diversity and structure for this clonal seed orchard. Total genomic DNA was extracted from fresh needle tissue with Plant Genomic DNA Extraction Miniprep System made by Viotechnology Corporation The results indicated that the clonal seed orchard of masson pine had higher genetic diversity. The average genetic diversity of the clonal seed orchard was 0.3169, the Shannon’s information index was 0.4813 respectively, and the percentage of polymorphic loci was 71.0%. Observed number of alleles (Na), effective number of alleles (Ne), Nei’s gene diversity (H), Shannon’s information index (I) and percentage of polymorphic loci (P) within population of Jiangxi, Hunan and Zhejiang were bigger than those of Guangdong, Guangxi, Anhui and Sichuan. Genetic distances among 8 populations were range from 0.0225 to 0.2175, whereas genetic identities were range from 0.8045 to 0.9777. 8 populations were clustered into 7 clusters, which showed that populations with similar latitude were clustered together and the clustering had nothing to do with geographic distributing. There was not significant correlation between genetic distance and geographic distance, while the correlation between genetic distance and latitude was more significant.
This study examines genetic variation in the Alabama hog sucker (Hypentelium etowanum) across river drainages in the southeastern United States using DNA sequencing of the mitochondrial cytochrome b gene. Tissue samples were collected from seven locations and DNA was extracted and amplified via PCR. Approximately 1150 base pairs of the cyt b gene were sequenced. Preliminary results found genetic variations between populations that are consistent with a previous study. The sequences from a new location in the Little Tallapoosa drainage were most closely related to those from the Chattahoochee drainage. This ongoing study aims to increase sampling to further resolve the genetic structure of H. etowanum across its range.
"Genomic approaches for dissecting fitness traits in forest tree landscapes"ExternalEvents
"Genomic approaches for dissecting fitness traits in forest
tree landscapes" presentation by Ciro De Pace, Università degli Studi della Tuscia, Viterbo, Italy
Identification and expression analysis of LEA gene family members in cucumber...asdasdas19
LEA (late embryogenesis abundant) proteins
are firstly discovered in seeds and then identified in vegetative tissues of different plant species. They are mainly
regulated under abiotic stress conditions. Although genome
wide studies of different gene family members have been
performed in cucumber, there is no such a study for LEA
genes. We have identified 79 LEA genes in the cucumber
genome. Based on phylogenetic analysis, CsLEA genes
could be classified into seven groups in which structural
motifs are relatively conserved. Tandem duplications play
an important role in cucumber genome for LEA gene
expansion. Orthologous and chromosomal relationships of
CsLEA genes were observed based on comparative mapping analysis with other species. The in silico micro-RNA
(miRNA) target analyses indicated that 37 CsLEA genes
were targeted by different miRNAs, especially mir854 and
mir414 are the most abundant identified ones. Public
available RNA-seq data were analyzed for expression
analysis of CsLEA genes in different tissues of cucumber
Concept and Use of Growth Hormones phd 701.pptxsouravranjan6
This document provides an overview of phytohormones and their roles in crop production. It discusses the main plant hormones (auxins, cytokinins, gibberellins, ethylene, abscisic acid, and brassinosteroids) and their functions in processes like germination, flowering, fruit ripening, and responses to stress. Several case studies demonstrate how applying phytohormones as seed treatments or foliar sprays can increase crop yields by promoting growth, biomass accumulation, and stress tolerance. The document is a useful introduction to the classification, biosynthesis, mechanisms of action, and agricultural applications of important phytohormone groups in field crops.
This document summarizes a study that evaluated callus induction from Calophyllum brasiliense (Cambes) explants and the production of anti-HIV calanolide metabolites. Seed and leaf explants were cultured on media with different plant growth regulators. Highest callus induction was achieved from seeds on media with BA and picloram, and from leaves on media with kinetin and NAA. Analysis found higher production of calanolide B and C in calluses from seeds compared to leaves. The study demonstrates the potential of tissue culture for producing these anti-HIV metabolites.
Similar to Association of Diaporthe longicolla with Black Zone Lines on Mature Soybean Plants (20)
2. PLANT HEALTH PROGRESS Vol. 16, No. 3, 2015 Page 116
plants were collected near or after maturation from two to three
arbitrarily selected soybean fields and examined for stems
containing zone lines (Table 1).
A study by Barr (1978) showed that Diaporthe spp. have
“stromatic tissues composed of prosenchymatous hyphae mixed
with substrate tissues in entostromatic regions, that often form
dark marginal zones above in epidermal tissues and frequently
deep in wood beneath perithecia or groups of perithecia.” More
recently, zone lines were characterized as a symptom produced by
either Macrophomina phaseolina (Tassi) Goid, Fusarium spp., or
Diaporthe spp. on soybean (Ghissi et al. 2014; Zaccaron et al.
2014). The objective of this study was to establish the identity of
the Diaporthe spp. associated with black zone lines produced on
the lower stems of mature soybean plants in Illinois, Indiana,
Iowa, Michigan, and South Dakota.
ISOLATION AND IDENTIFICATION OF THE CAUSAL AGENT
Stem samples (approximately 90) collected from the five states
were washed in tap water for 2 min; and approximately 1 cm-long
pieces were cut from the zone lines of the infected stems. The
pieces from each stem sample were surface-disinfested in sodium
hypochlorite (0.05%) and then ethanol (70%) for 1 min each,
rinsed in sterile distilled water four times, and blotted between
sterile filter paper. Four pieces were placed on potato dextrose
agar (PDA; Becton, Dickinson and Company, Franklin Lakes, NJ)
acidified to pH 4.5 with 85% lactic acid. Plates were incubated at
25
o
C for 7 to 10 days under fluorescent light with a photoperiod
of 12 h daily. Mycelial colonies appeared white, dense, and
floccose, and large black stromata were formed in concentric
patterns or scattered. Alpha conidia exuded from the pycnidial
ostiole in creamy-to-yellowish drops and were ellipsoid and
biguttulate with an average length of 6.6 μm and width of 2.1 μm.
Beta conidia and perithecia did not form on PDA. Ninety isolates
(one isolate per symptomatic plant) from the five states were
tentatively identified D. longicolla based on these morphological
characteristics (Santos et al. 2011). From the 90 symptomatic
stem samples collected from the five states, only D. longicolla
was isolated from the zone lines on the soybean stems. The 90 D.
longicolla isolates included 3 from Illinois, 20 from Indiana, 30
from Iowa, 30 from Michigan, and 7 from South Dakota.
To perform molecular identification of the causal agent, a total
of 10 representative isolates were selected and DNA was
extracted from the lyophilized mycelium scraped from the surface
of 10 day old cultures growing on PDA using the FastDNA Spin
Kit (MP Biomedicals, Solon, OH) (Table 1). The Diaporthe
isolates were identified to species by amplifying and sequencing
of the internal transcribed spacer (ITS) regions using primers
ITS1 and ITS2 (White et al. 1990). Approximately 600 bp of the
ITS region was amplified from the 10 isolates. Forward and
reverse sequences were assembled into contigs using BioEdit
software (v7.2.5) (Hall 1999). Analysis of the edited sequences
was performed using the Basic Local Alignment Search Tool
Nucleotide (BLASTn) searches at the GenBank database
(National Center for Biotechnology Information, http://www.ncbi.
nlm.nih.gov). A BLASTn search of GenBank performed for the
ITS sequences of the 10 isolates showed that the best match was
P. longicolla (syn. D. longicolla) strain STAM-35 (GenBank
Accession No. AY745021) from soybean in the United States
with identities ranging from 98% to 100%. Phylogenetic analyses
of the ITS sequences using maximum parsimony method placed
the 10 isolates in the group containing isolates of D. longicolla
and D. sojae from soybean in the United States (bootstrap value
of 85%; data not presented).
In order to establish a well-resolved phylogeny and clarify the
phylogenetic position, the 10 isolates were characterized by
phylogenetic analyses of two gene fragments, which included
EF1-α and ACT regions. The intron region of the EF1-α gene was
amplified using primers EF1-728F and EF1-986R (Carbone and
Kohn 1999). For the sequencing of the partial actin gene, frag-
ments of ACT gene were amplified using the primers ACT-512F
and ACT-783R (Carbone and Kohn 1999). All DNA samples were
sequenced (GenScript USA Inc., Piscataway, NJ) using the
respective primers. DNA sequences generated in this study have
been deposited under GenBank Accession Nos. KR815475 to
KR815494 and KT021543 to KT021552 (Table 1).
The EF1-α and ACT sequences of the 10 D. longicolla isolates
were aligned using the default parameters of ClustalX (Thompson
et al. 1997) and adjusted manually by visual examination using
the Molecular Evolutionary Genetics Analysis (MEGA) software
v6 (Tamura et al. 2013) prior to being exported as NEXUS files
for subsequent analyses. The outgroup Chrysoporthella hodges-
iana Gryzenhout and Wingfield (Isolate CMW9995) was obtained
from GenBank (GQ290152 for EF1-α sequence and GQ290170
for ACT sequence). The 45 sequences in the combined data set
(including the outgroup C. hodgesiana) comprised 688 bp of
aligned sequence. Phylogenies based on maximum parsimony
were inferred using MEGA, and heuristic searches for the most
parsimonious trees were conducted using the tree bisection-
TABLE 1
Diaporthe longicolla isolates used in this study, the genes sequenced, and the corresponding GenBank Accession Nos.
Isolatesa,b Locationb Species Identityc
GenBank Accession Numbers
ITS EF1-α ACT
IL1 Champaign County, IL D. longicolla KR815475 KR815485 KT021543
IL2 Champaign County, IL D. longicolla KR815476 KR815486 KT021544
IN1 Knox County, IN D. longicolla KR815477 KR815487 KT021545
IN2 Porter County, IN D. longicolla KR815478 KR815488 KT021546
IA1 Montgomery County, IA D. longicolla KR815479 KR815489 KT021547
IA2 Lee County, IA D. longicolla KR815480 KR815490 KT021548
MI1 Ingham County, MI D. longicolla KR815481 KR815491 KT021549
MI2 Ingham County, MI D. longicolla KR815482 KR815492 KT021550
SD1 Brookings County, SD D. longicolla KR815483 KR815493 KT021551
SD2 Miner County, SD D. longicolla KR815484 KR815494 KT021552
a A total of 10 isolates (two representative isolates from each state) were used for identification.
b
IL = Illinois, IN = Indiana, IA = Iowa, MI = Michigan, and SD = South Dakota.
c
Species identity was established based on sequence analysis of the internal transcribed spacer region (ITS), elongation factor subunit 1-α (EF1-α) and
actin (ACT) gene regions.
3. PLANT HEALTH PROGRESS Vol. 16, No. 3, 2015 Page 117
regrafting (TBR) algorithm (Nei and Kumar 2000) with search
level 1. The initial trees were obtained by random addition (10
replicates). Gaps were treated as missing data, and relative
support for branches was estimated with 1,000 bootstrap replica-
tions (Felsenstein 1985). Prior to the combined analyses, the
concordance of the two gene datasets was evaluated with the
partition-homogeneity test (Farris et al. 1994) implemented with
PAUP* v4.0b10 (Sinauer Associates, Inc., Sunderland, MA;
Swofford 2002) using 1,000 random repartitions (Felsenstein
1985), and with MAXTREES set to 5,000. The null hypothesis of
congruence was rejected if P < 0.001 (Dettman et al. 2003). For
the combined dataset, sequences were concatenated using
Mesquite v2.75 (Maddison and Maddison 2011).
The EF1-α and ACT genes were combined for characterization
of the 10 isolates based on the results of the partition-homo-
geneity test (P = 0.002), indicating that the EF1-α and ACT trees
reflect the same underlying phylogeny. The maximum parsimony
analysis for the combined dataset resulted in five most-parsi-
monious trees and a consensus tree was inferred from the five
trees (length = 262; Fig. 2). The consistency index, retention
index, and rescaled consistency index were 0.75, 0.92, and 0.74,
respectively. The D. longicolla isolates from the five states
formed a monophyletic group with the ex-type cultures CBS 179
and CBS 180 that was supported by a bootstrap value of 94%
(Fig. 2).
The cultural morphology and DNA sequence analysis of the 10
isolates from the five states corresponded to the description of D.
longicolla (syn. P. longicolla) (Santos et al. 2011).
PATHOGENICITY OF DIAPORTHE LONGICOLLA ISOLATES
The D. longicolla isolates were assayed for pathogenicity using
a modified toothpick inoculation method (Ghissi et al. 2014;
Keeling 1982). Seeds of a commercial soybean cultivar of relative
maturity 1.8 (Monsanto Company, St. Louis, MO) were sown into
a potting mix (Sunshine Mix #1, Sun Grow Horticulture Products,
Belleview, WA) in 7.5-liter, circular, plastic pots, one seed per pot
and grown for 4 weeks at 22°C in the greenhouse under a 14-h
photoperiod with a light intensity of 450 μE/m2
/s and watered
daily. The trial was conducted in a completely randomized design
with six replicates (pots) per treatment (D. longicolla isolate), and
the experiment was repeated once.
To obtain inoculum, the isolates were grown on PDA with
sterilized wood toothpicks placed on top of the media. After a
week of incubation, the toothpicks were removed from the PDA
and inserted 2 to 3 mm deep into the lower stems (above the
cotyledon) of soybean plants that were at approximate develop-
mental stages V3 to V4 (Fehr et al. 1971). The toothpicks and
stems were wrapped with Parafilm to avoid rapid dehydration. Six
plants were inoculated per isolate and one toothpick was inserted
per plant. Six control plants were treated by inserting one sterile,
non-infested toothpick into each stem. After inoculation, plants
were kept in the greenhouse for 7 weeks at 22°C with a 14-h
photoperiod. From the beginning of seed development (R5) plant
growth stage until maturation, the soybean plants were not
watered. Instead, the soybean plants were allowed to dry slowly
in the greenhouse at R5 developmental stage so that the fungal
inoculum on the inoculated plants would be dried in situ. At
developmental stage R7 (beginning maturity), soybean plants
were removed from the pots and washed, and examined for zone
lines on the soybean stems (Fig. 3). Upon examination, it was
observed that the occurrence of zone lines on the soybean stems
varied among the ten D. longicolla isolates. For example, zone
lines appeared on the exterior of the lower stems of the soybean
plants 7 weeks after inoculation with isolate IL1 compared to 8
weeks with the other isolates (Fig. 3). Zone lines were not
observed on any of the control plants.
To complete Koch’s postulates, D. longicolla was re-isolated
from the inoculated soybean plants, and the identity was
confirmed via sequencing of the ITS region using primers ITS1
and ITS2 (White et al. 1990). The pathogen was not recovered
from any of the control plants.
FIGURE 2
Consensus of five most-parsimonious trees (length = 262) resulting
from the alignment of 688 characters of the combined EF1-α and ACT
gene region of the Diaporthe longicolla isolates using the maximum-
parsimony analyses. The consistency index, retention index, and
rescaled consistency index were 0.75, 0.92, and 0.74, respectively.
Bootstrap values greater than 50 are shown. D. longicolla isolates
recovered from soybean fields in the five states have state labels (IL =
Illinois, IN = Indiana, IA = Iowa, MI = Michigan, and SD = South Dakota).
The ex-type cultures CBS 179 and CBS 180 are labelled ‘dl’ for D.
longicolla. Chrysoporthella hodgesiana (isolate CMW9995) was used as
the outgroup (National Center for Biotechnology Information Accession
No. GQ290152 for EF1-α sequence and GQ290170 for ACT sequence).
Evolutionary analyses were conducted in MEGA6 (Tamura et al. 2013).
4. PLANT HEALTH PROGRESS Vol. 16, No. 3, 2015 Page 118
IMPORTANCE AND IMPACT
This study demonstrated that the causal fungus producing zone
lines on the lower stems of mature soybean plants in Illinois,
Indiana, Iowa, Michigan, and South Dakota is D. longicolla. The
identification of the causal agent was confirmed by morphology,
DNA sequence analyses of the ITS, EF-1α, and ACT gene
regions, and completion of Koch’s postulates. The phylogenetic
tree generated using the maximum parsimony method by
combining EF-1α and ACT dataset clustered the D. longicolla
isolates from the five states with the D. longicolla ex-type
cultures CBS 179 and CBS 180 forming a monophyletic group
(94% bootstrap support; Fig. 2). Previously, Ghissi et al. (2014)
identified D. phaseolorum var. sojae to be associated with zone
lines on soybean plants based on morphological characteristics.
Zaccaron et al. (2014) associated D. longicolla, M. phaseolina,
and Fusarium spp. with the zone lines observed on tap roots of
the soybean plants based on the recovery of the three fungal
pathogens from the zone lines. However, in this study, only D.
longicolla was isolated from the zone lines on the soybean stems
sampled from the five states. To the best of our knowledge, this is
the first study demonstrating that D. longicolla is associated with
black zone lines on mature soybean plants in Illinois, Indiana,
Iowa, Michigan, and South Dakota.
In this study, the pathogenicity experiment to produce zone
lines by D. longicolla on soybean plants was performed using a
modified protocol of Ghissi et al. (2014) and Keeling (1982). The
protocol was modified from R5 developmental stage to stop
watering the soybean plants and subject them to slow drying in
the greenhouse; given previous studies suggested that zone lines
may be formed in host tissues from the slow drying of the host
plant (Nikandrow 1989; Hubert 1924). However, no watering
during R5 development stage accelerated the natural senescing of
the soybean plants in this experiment. This also caused the
soybean plants to break off as they approached the R7 develop-
ment stage and the experiment had to be terminated. As a result of
this, zone line and stroma were only observed on the exterior of
the lower stem of inoculated plants (Fig. 3).
In the pathogenicity study, differences were observed among D.
longicolla isolates for the amount of time to produce zone lines
and stroma on soybean stems under greenhouse conditions. For
instance, Isolate IL1 produced zone lines and stroma on the
exterior of the lower stems of soybeans in 7 weeks as compared to
the other isolates in 8 weeks indicating that the D. longicolla
isolates may possibly vary in virulence. Although it is likely that
virulence of the D. longicolla isolates or environmental condi-
tions may have influenced pathogen establishment on the host and
the rate at which the pathogen can grow through the host tissue,
this was beyond the scope of our study. However, recent studies
by Zaccaron et al. (2014) have indicated that the production of
zone lines in senesced soybean plants seems to be affected by
cultivar type and location. Further research is warranted to
understand the role of zone lines during the interaction between
D. longicolla and soybean.
D. longicolla primarily causes Phomopsis seed decay of
soybean in areas where high temperatures and high humidity
occur during crop maturation (Hobbs et al. 1985). In Illinois and
other parts of the North Central United States, Phomopsis seed
decay is known to be endemic on soybeans seeds (Sinclair 1993).
Although the infection of soybean seeds with D. longicolla has
resulted in significant economic losses, especially in the mid-
southern region of the United States (Hepperly and Sinclair
1978), there is very little information on stem diseases caused by
D. longicolla. In North Dakota and South Dakota, soybean stem
canker-affected samples were collected during late-season disease
surveys and D. longicolla was isolated from these stems along
with D. caulivora (Mathew et al. 2015b; A. Gebreil and F.
Mathew, unpublished data). Pathogenicity studies by Gebreil et
al. (2015) suggest that D. longicolla may be more aggressive than
D. caulivora on soybean stems. Although D. longicolla has been
identified as a stem pathogen on soybean in North Dakota and
South Dakota (Mathew et al. 2015b; Gebreil et al. 2015), there is
no information on associated yield loss and seed quality.
In summary, the formation of zone lines on the stems of mature
soybean plants is diagnostic of the presence of D. longicolla.
Previously, zone lines were characterized as produced by the
Charcoal rot pathogen (M. phaseolina), Fusarium spp., or
Diaporthe spp. (Ghissi et al. 2014; Zaccaron et al. 2014; T.
Hughes, personal communication), when they were observed on
soybean as the plants approached maturity. Studies by Nikandrow
(1989) and Campbell (1933) suggest that zone lines are the cross-
sectional view of melanized fungal tissue embedded in the host.
On grapevine (Vitis vinifera L.), canes have zone lines when
perithecia (fruiting bodies) produced by the causal fungus (D.
viticola Nitschke) are present (Scheper et al. 2000). On soybean,
it is not clear if the formation of zone lines is related to the
production of perithecia by D. longicolla and this warrants further
research. Although there is limited information available on the
biology of D. longicolla as a stem pathogen and subsequent
disease development, agricultural personnel and crop consultants
in the soybean industry should be aware that the black zone lines
are produced by D. longicolla on soybean plants.
ACKNOWLEDGMENTS
This research was supported by the South Dakota Soybean Research
and Promotion Council (Sioux Falls, SD), the South Dakota Agricultural
Experimental Station (Hatch Project H527-14), and the North Central
Soybean Research Program (Ankeny, IA). We thank Warren Pierson,
Nolan Anderson, Connie Tande, Kay Ruden, Paul Okello, Luke
Hyronimus, Scott Mages, Brian Kontz, and Alec Weber for their
assistance in this study.
FIGURE 3
Zone lines produced by (A) Diaporthe longicolla on the exterior of the
lower stem of a soybean plant under natural conditions and (B) by D.
longicolla isolate IL1 on the lower stem of a soybean plant in the
greenhouse (Photo credit: Connie Tande).
5. PLANT HEALTH PROGRESS Vol. 16, No. 3, 2015 Page 119
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