High Polymorphism Information Content Research Articles

Mining expressed sequence tag (EST) microsatellite markers to assess the genetic differentiation of five Hynobius species endemic to Taiwan

Taiwan harbors five endemic species of salamanders (Hynobius spp.) that inhabit distinct alpine regions, contributing to population fragmentation across isolated “sky islands”. With an evolutionary history spanning multiple glacial-interglacial cycles, these species represent an exceptional paradigm for exploring biogeography and speciation. However, a lack of suitable genetic markers applicable across species has limited research efforts. Thus, developing cross-amplifying markers is imperative. Expressed sequence-tag simple-sequence repeats (EST-SSRs) that amplify across divergent lineages are ideal for species identification in instances where phenotypic differentiation is challenging. Here, we report a suite of cross-amplifying EST-SSRs from the transcriptomes of the five Hynobius species that exhibit an interspecies transferability rate of 67.67%. To identify individual markers exhibiting cross-species polymorphism and to assess interspecies genetic diversity, we assayed 140 individuals from the five species across 84 sampling sites. A set of EST-SSRs with a high interspecies polymorphic information content (PIC = 0.63) effectively classified these individuals into five distinct clusters, as supported by discriminant analysis of principal components (DAPC), STRUCTURE assignment tests, and Neighbor-joining trees. Moreover, pair-wise FST values > 0.15 indicate notable between-cluster genetic divergence. Our set of 20 polymorphic EST-SSRs is suitable for assessing population structure within and among Hynobius species, as well as for long-term monitoring of their genetic composition.

Individual Identification and Assessment of Genetic Diversity Using Microsatellite Markers in Racing Pigeons Raised in Turkiye

The implementation of a swift and economical molecular genetic approach, ensuring both efficacy and cost-effectiveness and facilitating population certification, is of utmost significance for breeders and the conservation of Turkiye's native pigeon biodiversity. In this study, we aimed to examine the genetic structure of racing pigeons (Columba livia domestica) raised in Turkiye using a genetic marker panel consisting of eight short tandem repeat (STR) loci. For this purpose, DNA was isolated from the shed feathers of 216 pigeons. Genomic DNA was amplified using the multiplex allele-specific PCR and subsequent capillary electrophoresis with ABI PRISM 3130XL Genetic Analyzer. Next, PCR products were analyzed in the GeneMapper Software program (Applied Biosystems). For parent testing, paternity index (PI), combined paternity index (CPI), and cumulative probability of paternity (CPP) were calculated. Furthermore, population genetic diversity was evaluated using heterozygosity (He), polymorphism information content (PIC), and Hardy–Weinberg equilibrium (HWE) testing. Results revealed that the total number of alleles is 81 and the number of alleles per locus varies between 4 and 19. The similarity rate between parent and offspring was calculated as 99.99% and above. Since no pedigree information was given when the samples were analyzed, obtaining this similarity ratio demonstrates the reliability of the panel. He values range from 0.362 to 0.919, and the PIC values range from 0.295 to 0.909. Loci PG-1, PG-2, and PG-3 show significant genetic diversity, with moderate to high PIC values reflecting varied allele frequencies in the population. Consequently, the set of seven STR markers (+ one sex marker) can be applied to identify and confirm parentage on a regular basis, thereby facilitating efficient breeding programs and ensuring genetic diversity conservation. This panel enables efficient pedigree analysis and gender determination, optimizing cost-effectiveness. The methodology presented in this study is ideal for pedigree analysis and breed certification in the Turkish pigeon breeding industry. Consequently, we affirm that the study data carries considerable national importance.

Phenotypic and Molecular-Markers-Based Assessment of Jamun (Syzygium cumini) Genotypes from Pakistan

Jamun plant displays enormous diversity throughout Pakistan, which necessitates its screening, evaluation, and validation to document elite genotypes having better traits for the benefit of the fruit industry and farmers. Surveys were made in natural Jamun habitats across Punjab, Pakistan, and genotypes were marked based on visual diversity of trees and fruits. In total, 60 Jamun genotypes were selected for characterization based on phenotypic and genetic markers. Phenotypic characters related to trees, leaf, and flower along with fruit qualitative traits were assessed in situ. Results revealed significant diversity with high (>25%) coefficient of variance values and the first two components of correspondence analysis exhibited 41.71% variation among genotypes. A strong association was observed among traits like upright tree and round fruit shape (0.74), bluish-colored fruit and pinkish pulp (0.85), and elliptic-shaped fruit with low fruit waxiness (−0.72). Leaves of phenotypically characterized plants were brought to Wheat Biotechnology Lab., University of Agriculture, Faisalabad, Pakistan, where Jamun genotypes were investigated genetically using Random Amplified Polymorphic DNA (RAPD) and Inter Simple Sequence Repeat (ISSR) markers. A total of 132 bands were scored, of which 108 were polymorphic, corresponding to almost 81% polymorphism among collected genotypes. High polymorphism information content values were observed against RAPD (0.389) and ISSR (0.457) markers. Genotypes were compared in relation to genetic markers, which exhibited that almost 86% of genetic variability was attributed to differences among accessions, while 14% of variation was due to differences between collections of different areas. Findings of this study confirmed wide phenotypic and genetic distinctness of Jamun in Pakistan that can aid breeders for marker-assisted selection and germplasm enhancement for future crop improvement programs.

Genetic diversity of maize resources revealed by different molecular markers

AbstractMaize (Zea mays L.) is the third most important cereal crop in the world because of its nutritional value and industrial benefits. Molecular markers are used mainly by the breeders to study the genetic variability of genotypes and its application in the breeding process. Two types of molecular markers, 10 random amplified polymorphic DNA (RAPD) primers and 10 start codon target (SCoT) primers, were assayed to determine the genetic diversity of 25 Slovak maize lines and 25 maize cultivars. A high level of polymorphism was found with both RAPD and SCoT markers, which was confirmed by high average polymorphism information content (PIC) values using both techniques. The efficiency of individual marker techniques in the detection of genotype diversity can be compared by calculating the marker index (MI), detecting diversity index (DDI), discriminating power, resolving power (RP) and other indices. A higher MI (11.788), DDI (2.358) and RP (53.08) value was achieved by the SCoT technique compared to the RAPD method. Three joint dendrograms and PCoA plots constructed based on RAPD, SCoT and both methods combined confirmed the unambiguous separation of maize lines and cultivars from each other. The results obtained from the RAPD and SCoT analysis can be used for the selection of potentially suitable biological sources for further marker-assisted breeding.

Aromatic rice (Oryza sativa L.) landraces of Indo‐Gangetic Plains of India harbor rich genetic diversity

AbstractAromatic rice (Oryza sativa L.) germplasm conserved in global genebanks needs intensive characterization to assess genetic diversity and population structure for crop improvement. We analyzed a panel of 92 accessions of non‐Basmati aromatic rice landrace germplasm with 23 phenotypic traits and 48 microsatellite markers. The landrace accessions displayed a huge phenotypic diversity for traits, namely, culm length, flag‐leaf length, panicle‐bearing tillers (PBT), plant height, panicle length, peduncle length, panicle exsertion, grain length, grain width, hull color, and 1000‐grain weight (TGW). Phenotypic clustering revealed three major clusters and five principal components explained 81.24% of the variance. TGW was associated positively with grain length, grain width, peduncle length, and grain yield but negatively with culm length and PBT. A total of 528 microsatellite alleles were detected, with 11.0 alleles per locus, revealing the presence of high polymorphism. Genetically, four subpopulations were detected based on structure analysis. In the germplasm, high levels of gene flow (Nm = 1.51) and gene diversity (HE = 0.83) were found. Among the subpopulations, we discovered high levels of inbreeding (FIS = 0.92) and moderate genetic differentiation (FST = 0.14). High polymorphism information content (0.806) along with a high level of molecular variance among genotypes (79%) was detected. A large number of multiple alleles were detected in numerous landrace accessions, revealing the presence of heterogeneity and heterozygotes in the landrace germplasm. The results of the present study are vital for the utilization of landrace diversity in the genetic improvement programs designed for rice quality improvement and consumer preference.

Genetic diversity analysis and fingerprinting of 175 Chimonanthus praecox germplasm based on SSR molecular marker

The elucidation of resources pertaining to the Chimonanthus praecox varieties and the establishment of a fingerprint serve as crucial underpinnings for advancing scientific inquiry and industrial progress in relation to C. praecox. Employing the SSR molecular marker technology, an exploration of the genetic diversity of 175 C. praecox varieties (lines) in the Yanling region was conducted, and an analysis of the genetic diversity among these varieties was carried out using the UPDM clustering method in NTSYSpc 2.1 software. We analyzed the genetic structure of 175 germplasm using Structure v2.3.3 software based on a Bayesian model. General linear model (GLM) association was utilized to analyze traits and markers. The genetic diversity analysis revealed a mean number of alleles (Na) of 6.857, a mean expected heterozygosity (He) of 0.496 3, a mean observed heterozygosity (Ho) of 0.503 7, a mean genetic diversity index of Nei՚s of 0.494 9, and a mean Shannon information index of 0.995 8. These results suggest that the C. praecox population in Yanling exhibits a rich genetic diversity. Additionally, the population structure and the UPDM clustering were examined. In the GLM model, a total of fifteen marker loci exhibited significant (P < 0.05) association with eight phenotypic traits, with the explained phenotypic variation ranging from 14.90% to 36.03%. The construction of fingerprints for C. praecox varieties (lines) was accomplished by utilizing eleven primer pairs with the highest polymorphic information content, resulting in the analysis of 175 SSR markers. The present study offers a thorough examination of the genetic diversity and SSR molecular markers of C. praecox in Yanling, and establishes a fundamental germplasm repository of C. praecox, thereby furnishing theoretical underpinnings for the selection and cultivation of novel and superior C. praecox varieties, varietal identification, and resource preservation and exploitation.

Genetic Relatedness in Elite Cultivars of Moth Bean using Morpho-agronomic and Molecular Markers

Background: Moth bean is a very nutritive and hardy crop, especially for resource poor-arid agriculture. The limited use of cultivars in moth bean breeding resulted into narrow genetic base. This study aimed to diversify the conical genetic base and developimproved cultivars, through molecular (RAPD) and morpho-agronomic characterization of commercially elite moth bean cultivars. Methods: Elite moth bean cultivars (15) were evaluated for 11 morpho-agronomic traits. Eleven RAPD markers were used to amplify genomic DNA and perform molecular characterization.Clustering approach was used for grouping of the cultivars. Result: RAPD markers revealed high polymorphism in 15 elite moth bean cultivars. The polymorphism information content (PIC) values varied within the range of 0.45 to 0.88, with an average of 0.77. The number of alleles at different loci ranged from 4 to 16, with an average of 10 alleles per locus. When employing UPGMA-based cluster analysis, utilizing 11 morpho-agronomic traits, the cultivars were grouped into four clusters. However, when RAPD markers were used, the cultivars were grouped into five clusters. The Jaccard’s similarity coefficient and Manhattan dissimilarity coefficient fell within the ranges of 0.18 to 0.70 and 0.02 to 0.42, respectively. These values signify the degree of genetic variability within the cultivars. Furthermore, a Mantel test was conducted to examine the correlation between agronomic traits and the RAPD-based matrix. The results showed a negative correlation, but it was not statistically significant.The high PIC values and the successful amplification of multiple loci demonstrate the efficacy of RAPD markers in assessing genetic diversity in moth beans. The study revealed enormous genetic variation among cultivars and crosses can be attempted between cultivars of different groups to create better recombinants in moth bean breeding programmes.

Revealing Y-STR Diversity of Koli Populations (Gujarat) by Studying 23 Y-STR Loci

Genetic analysis of Y-STR loci is pivotal for forensic libraries and genetic analysis. The Koli population in Gujarat, India, however, lacks such genetic characterization. This study aims to develop an allele frequency database for 23 Y-STR loci in the Koli population, examining forensic parameters and assessing genetic connections with neighboring tribes. A total of 153 unrelated Koli males were genotyped using the PowerPlex®Y23 multiplex commercial kit. We identified 117 distinct haplotypes. The Haplotype Diversity (HD) and Discrimination Capacity (DC) for the 23 Y-STR loci were 0.993 and 0.8034, respectively. DYS385b locus exhibited the highest allele variability (10 alleles), whereas DYS391, DYS389I, and DYS437 showed the least (4 alleles each). The highest Polymorphic Information Content (PIC) was observed in DYS385b (0.775), with the lowest in DYS391 (0.386). The dominant haplogroup R1a accounted for 45% of the population. Comparative analysis with other Indian populations from YHRD revealed two distinct clusters, placing the Koli population in cluster 2, indicating significant genetic similarity within this group. This inaugural study of Y-STRs in the Koli population demonstrates the utility of the Y23 kit in male identification, highlighted by substantial haplotype diversity and discrimination capacity.

Microsatellite imputation using SNP data for parentage verification in four Italian sheep breeds.

Microsatellite markers (MS) have been widely used for parentage verification in most of the livestock species over the past decades mainly due to their high polymorphic information content. In the genomic era, the spread of genotype information as single-nucleotide polymorphism (SNP) has raised the question to effectively use SNPs also for parentage testing. Despite the clear advantages of SNP panels in terms of cost, accuracy, and automation, the transition from MS to SNP markers for parentage verification is still very slow and, so far, only routinely applied in cattle. A major difficulty during this transition period is the need of SNP data for parents and offspring, which in most cases is not yet feasible due to the genotyping cost. To overcome the unavailability of same genotyping platform during the transition period, in this study we aimed to assess the feasibility of a MS imputation pipeline from SNPs in four native sheep dairy breeds: Comisana (N = 331), Massese (N = 210), Delle Langhe (N = 59) and Sarda (N = 1003). Those sheep were genotyped for 11 MS and with the Ovine SNP50 Bead Chip. Prior to imputation, a quality control (QC) was performed, and SNPs located within a window of 2 Mb from each MS were selected. The core of the developed pipeline was made up of three steps: (a) storing of both MS and SNP data in a Variant Call Format file, (b) masking MS information in a random sample of individuals (10%), (c) imputing masked MS based on non-missing individuals (90%) using an imputation program. The feasability of the proposed methodology was assessed also among different training - testing split ratio, population size, number of flanking SNPs as well as within and among breeds. The accuracy of the MS imputation was assessed based on the genotype concordance as well as at parentage verification level in a subset of animals in which assigned parents' MS were available. A total of 8 MS passed the QC, and 505 SNPs were located within the ±2 Mb window from each MS, with an average of 63 SNPs per MS. The results were encouraging since when excluding the worst imputed MS (OARAE129), and regardless on the analyses performed (within and across breeds) for all breeds, we achieved an overall concordance rate over 94%. In addition, on average, the imputed offspring MS resulted in equivalent parentage outcome in 94% of the cases when compared to verification using original MS, highlighting both the feasibility and the eventual practical advantage of using this imputation pipeline.

CoreSNP: an efficient pipeline for core marker profile selection from genome-wide SNP datasets in crops

BackgroundDNA marker profiles play a crucial role in the identification and registration of germplasm, as well as in the distinctness, uniformity, and stability (DUS) testing of new plant variety protection. However, selecting minimal marker sets from large-scale SNP dataset can be challenging to distinguish a maximum number of samples. Results: Here, we developed the CoreSNP pipeline using a “divide and conquer” strategy and a “greedy” algorithm. The pipeline offers adjustable parameters to guarantee the distinction of each sample pair with at least two markers. Additionally, it allows datasets with missing loci as input. The pipeline was tested in barley, soybean, wheat, rice and maize. A few dozen of core SNPs were efficiently selected in different crops with SNP array, GBS, and WGS dataset, which can differentiate thousands of individual samples. The core SNPs were distributed across all chromosomes, exhibiting lower pairwise linkage disequilibrium (LD) and higher polymorphism information content (PIC) and minor allele frequencies (MAF). It was shown that both the genetic diversity of the population and the characteristics of the original dataset can significantly influence the number of core markers. In addition, the core SNPs capture a certain level of the original population structure.ConclusionsCoreSNP is an efficiency way of core marker sets selection based on Genome-wide SNP datasets of crops. Combined with low-density SNP chip or genotyping technologies, it can be a cost-effective way to simplify and expedite the evaluation of genetic resources and differentiate different crop varieties. This tool is expected to have great application prospects in the rapid comparison of germplasm and intellectual property protection of new varieties.

Characterization of heterogeneity in popular rice landrace through field and molecular evaluation

ContextIdentification and purification of the landrace Chittimuthyalu using stability and molecular heterozygosity can be useful for rice diversity. ObjectivesThe present study reports the characterization of 12 genotypic variants of Chittimuthyalu in terms of (i) variation and correlation among agromorphological, grain quality and nutrient traits, viz., Zn and Fe; (ii) stability across years; and (iii) characterization and heterozygosity. MethodsA total of twelve genotypic variants were identified based on field-level agromorphological observations in the Chittimuthyalu landrace. For identification of a stable genotypic variant for grain Zn, genotype by environment (G × E) interaction was analyzed across three years using additive main effects and multiplicative interaction (AMMI) model and genotype main effects plus G × E (GGE) model for multienvironment data analysis along with principal component analysis (PCA). Correspondingly, twenty-five high polymorphism information content (PIC) RM markers were used for the identification of molecular heterozygosity in landrace. ResultsStepwise regression analysis of phenotype revealed the interdependence of zinc in polished rice (ZPR), iron in polished rice (IPR), and single plant yield (SPY) and the positive association of grain Zn with IPR and SPY. Based on GGE and AMMI analyses of the variants, MCM3 was found to be stable across the environments for ZPR. Out of 46% polymorphic RM markers, seven markers (RM10149, RM13347, RM21749, RM13263, RM10361, RM12276 and RM474) were heterozygous. Heterozygous alleles were observed in eight variants (MCM2, MCM3, MCM5, MCM6 MCM7, MCM8, MCM10 and MCM12). The polymorphic information content (PIC) of the markers ranged from 0.58 to 0.88, with an average of 0.64. The percentage of the most common allele frequencies ranged from 4.2 to 95.8 across the markers. RM13347 showed nine markertrait associations with P ≤ 0.05 and R2 ranging from 2.3% to 14.3% with a mean of 7.74%. ConclusionsThe present study has revealed the inherent heterogeneity in landraces in terms of their agromorphological, yield-related and nutritional quality traits. A promising genetic stock (MCM3) was identified as a donor for high grain ZPR and was registered as genetic stock. ImplicationsThe identified genetic stock is being shared with breeders in the breeding programme and farmers for the multiplication of high grain Zn rice.

Microsatellite-Based Genetic Diversity Analysis and Population Structure of Proso Millet (Panicum miliaceum L.) in Kazakhstan

Proso millet is an important allotetraploid cereal crop; however, it is the least studied species of the Poaceae family, as it is an under-utilized crop. Genomic resources for proso millet are very limited compared to major crops. An understanding of the genetic relationships among germplasm resources is important for future breeding endeavors. In the present study, simplesequence repeat (SSR) markers were employed to assess the polymorphism and genetic diversity of 100 millet accessions from different countries, which were tested in the dry steppe zone conditions of the Akmola region from 2020 to 2022. The use of 20 SSR markers detected a total of 47 alleles, with an average allele number of 2.35 per locus among these proso accessions. Nine of them were polymorphic among the genotypes, which suggests that these SSR markers can be used for genetic studies. The results showed a moderate level of polymorphism information content (PIC) that averaged at 0.424, ranging from 0.125 to 0.795. The markers SSR-67, SSR-82, SSR-85 and SSR-109 showed high PIC values of 0.536, 0.756, 0.795 and 0.758, respectively. Markers SSR 85 and SSR 86 significantly correlated to agronomic traits, such as productive tillering (PT) and grain yield (GY). The genetic structure, UPGMA cluster and PCoA assay indicated that the accessions that originated from Central Asia had higher genetic diversity. Based on structure (K = 3), all the accessions were divided into three groups, where the gene pool that originated from Central Asia wasdetected in all three clusters. Based on a principal component analysis (PCA), the accessions of Central Asian origin were genetically closer to the North Asian group.

Population structure and genetic differentiation analyses reveal high level of diversity and allelic richness in crop wild relatives of AA genome species of rice (Oryza sativa L.) in India.

Crop wild relatives (CWRs) are vital sources of variation for genetic improvement, but their populations are few in genebanks, eroded in natural habitats and inadequately characterized. With a view to explore genetic diversity in CWRs of AA genome rice (Oryza sativa L.) species in India, we analyzed 96 accessions of 10 Oryza species by using 17 quantitative traits and 45 microsatellite markers. The morpho-quantitative traits revealed a high extent of phenotypic variation in the germplasm. Diversity index (H') revealed a high level of within-species variability in O. nivara (H' = 1.09) and O. rufipogon (H' = 1.12). Principal component (PC) analysis explained 79.22% variance with five PCs. Among the traits related to phenology, morphology, and yield, days to heading showed strong positive association with days to 50% flowering (r = 0.99). However, filled grains per panicle revealed positive association with spikelet fertility (0.71) but negative with awn length (- 0.58) and panicle bearing tillers (- 0.39). Cluster analysis grouped all the accessions into three major clusters. Microsatellite analysis revealed 676 alleles with 15.02 alleles per locus. High polymorphism information content (PIC = 0.83) and Shannon's information index (I = 2.31) indicated a high level of genetic variation in the CWRs. Structure analysis revealed four subpopulations; first and second subpopulations comprised only of O. nivara accessions, while the third subpopulation included both O. nivara and O. rufipogon accessions. Population statistics revealed a moderate level of genetic differentiation (FST = 0.14), high gene diversity (HE = 0.87), and high gene flow (Nm = 1.53) among the subpopulations. We found a high level of molecular variance among the genotypes (70%) and low among populations (11%) and within genotypes (19%). The high level of molecular and morphological variability detected in the germplasm of CWRs could be utilized for the improvement of cultivated rice.

Morphological Variability and Molecular Diversity in Blackgram (Vigna mungo (L.) Hepper) Genotypes Using SSR Markers

A study on the Morphological variability of 25 Blackgram genotypes was carried out in which molecular diversity of 15 genotypes of Blackgram was studied by using 8 SSR markers. The GCV was lower than PCV for all traits, however, there was narrow difference in PCV and GCV, indicating the effect of environment in lesser frequency. The high seed yield was recorded in the genotypes PLU-429(8.06gm) followed by PL-416(7.37gm) and VBN-8(7.29gm). Among all the factors PC 1 to PC 8, the PC 1 (40.610) accounted maximum proportion of variability in the set of all variables. Association studies revealed that there was high and significant positive amount of genotypic as well as phenotypic correlation coefficient for traits viz., primary branches per plant, number of clusters per plant, number of pods per plant, harvest index. Path analysis revealed high positive direct effect was contributed by primary branches per plant, number of clusters per plant, number of pods per plant, harvest index both at genotypic and phenotypic levels. The Polymorphism information content PIC values of 8 SSR markers valued from 31 percent to 71 percent with mean of 51 percent. BG 18 exhibited highest PIC 71 percent and highest heterozygosity of 62 percent. As per Dissimilarity coefficient, 15 genotypes are grouped into five clusters, Cluster II &amp; V has highest 4 genotypes while in compare with Cluster I, III &amp; IV. SSR marker BG18 has highest number of alleles 5. Shannon’s information index(I) ranged from 1.465 for BG18 to 0.611 for BG07. Fixation index is ranged from 1.000 to -0.111 with mean 0.696. SSRs with high polymorphism information content successfully assisted in the differentiation of genotypes in this study.

Microsatellite markers’ based molecular divergence among the breeding lines of aromatic marigold (Tagetes minuta L.)

Aromatic marigold (Tagetes minuta L.) is an essential oil-bearing plant of industrial value. The essential oil obtained from the plant has high demand in the flavor and perfume industry. The present study investigates the molecular diversity patterns using microsatellite markers in 126 breeding lines representing three groups (higher, intermediate, and lower biomass) based on phenotypic characterization. These lines are being maintained through repeated selfing. A total of 503 alleles with an average of 16.23 alleles per locus were detected at 31 SSR loci over all the genotypes. The average most common and rare alleles were 3.35 and 12.87, respectively, over all the studied loci. The high polymorphic information content (0.88) and genetic diversity (0.89) indicate high allelic diversity among the genotypes studied. Ten genotype-specific markers were identified. Significant molecular variances have been observed among the groups (3 %), among individuals (72 %), and within individuals (26 %). The unweighted neighbor-joining clustering based on a simple matching dissimilarity index grouped all 126 genotypes into five major clusters. The clustering pattern primarily follows the biomass-specific distribution of populations. The principle coordinate analysis depicts considerable variations and again confirms the biomass-specific grouping of genotypes. The highly significant positive association between genotypic and phenotypic distances indicates the importance of these marker loci for future marker-assisted selection and association mapping studies.

Assessment of Genetic Diversity and Protein Content of Scandinavian Peas (Pisum sativum)

We produced homogeneous lines of 227 pea accessions from the Nordic Genetic Resource Center via single seed descent. The genetic diversity among these, mostly Scandinavian accessions, was investigated using three microsatellite markers, A9, AC58 and AA5. The microsatellites were highly informative and separated 153 of 194 accessions on a Neighbor Joining topology. The high polymorphism information content (PIC) values between 0.87 and 0.91 indicated that the gene bank material contains a large number of pea accessions with different breeding histories. The peas were grown in the field for two years and seed protein content showed variation between 9.3% and 34.1% over the years and accessions, respectively. The mean thousand seed weight was 152.05 g. More than 10 accessions had a protein content above 28%, showing that the collection has potential as breeding nursery for high-protein pea cultivars.

Efficiency of RAPD and SCoT Markers in the Genetic Diversity Assessment of the Common Bean.

Knowledge about the genetic diversity of the available common bean germplasm can help breeders properly direct the choice of genetic material in the breeding process. The aim of the present work was to estimate the usefulness of 10 RAPD and 10 SCoT markers in genetic diversity detection among 33 common bean genotypes. Both molecular marker systems were able to generate high levels of polymorphism in the genetic material, which was supported by the relatively high polymorphic information content (PIC) values observed for the used markers. The Diversity Detection Index (DDI) and Marker Index (MI) were used to compare the effectiveness of RAPD and SCoT markers. For both techniques, high values of MI and DDI were calculated, representing their effectivity. The SCoT markers showed higher values of the parameters used (MI = 7.474, DI = 2.265) than the RAPD markers (MI = 5.323, DDI = 1.612), indicating their higher efficiency in the detection of molecular variability. Three constructed dendrograms and PCoA plots were created using RAPD and SCoT, and both methods combined confirmed sufficient separation of the bean genotypes from each other. At the same time, a higher efficiency of SCoT markers compared to RAPD markers in the detection of the genetic diversity of beans was also proven. The results may be of future interest in the choice of genetically distant material for breeding purposes.

GENETIC PURITY AND DIVERSITY ASSESSMENT OF PARENTAL CORN INBRED LINES USING SSR MARKERS FOR PHILIPPINE HYBRID BREEDING

Accurate data and relevant insights on parental corn inbred lines' and hybrids’ genetic purity and diversity are essential for hybrid development and seed production. Here, the genetic purity and diversity of 19 parental yellow corn inbred lines were assessed using SSR markers. A total of 91 SSR markers were utilized, of which 61 were highly polymorphic and had high polymorphism information content value (PIC = 0.379). Genetic purity and diversity parameters were calculated from the generated SSR marker data. Observed pairwise genetic distances ranging from 0.257 to 0.808 implied high genetic dissimilarities among the accessions. Cluster analysis separated the parental lines into three distinct subclusters, which can potentially be a basis for generating heterotic groupings among the parental lines. Eight (8) out of the 19 parental lines showed considerable residual heterozygosity of ≤10%. Inbred line CML 431 displayed complete homozygosity across all 61 SSR markers. Inbred lines that have residual heterozygosity of &gt;15% need purification through further breeding and selection. Out of the resultant F1 hybrids analyzed, only four (4) showed genetic impurity of ≤10%. It may be attributed to the intrinsic genetic impurity of parental line CML 452 (21.67%). In contrast, hybrids generated from two genetically pure parents (e.g., CML 431 and CML 575) showed low to no off-types. Overall, genetic purity and diversity determination of promising parental lines can be valuable for future yellow corn breeding programs in the Philippines.

Assessment of genetic diversity in cowpea (Vigna unguiculata L.) genotypes by using RAPD markers

The present study was carried out at the department of Plant Biotechnology, College of Agricultural Biotechnology Latur, which aimed to study or assess the genetic diversity in eight different Cowpea genotypes by using the RAPD markers. This is important for cowpea genetic improvement and for the overall management of cowpea germplasm. Eight Cowpea genotypes namely GDVC-2, GC-3, GC-4, GC-6, Gangotri, Pusa Komal, Arka Garima, and VU-5 were assessed by using two RAPD markers (RPI-07 and RPI-08). In RAPD analysis, total of 20 amplicons were generated, among them 19 were polymorphic with 88.8% average polymorphism. RPI-7 and RPI-8 showed 88.8% and 100% polymorphism respectively. The similarity coefficient values ranged from 0.350% to 0.950% indicating high degree of genetic variation. The highest genetic similarity i.e. 0.857% was recorded between Pusa Komal and GC-6 and least genetic similarity i.e. 0.125% was observed in between VU-5 and GC-4. Two screened primers are found to be polymorphic (94.4%) with high polymorphic information content (PIC) which efficiently discriminating the varieties at molecular level.

Assessment of Genetic Diversity of Local Coffee Populations in Southwestern Saudi Arabia Using SRAP Markers

Coffea arabica, a member of the Rubiaceae family, is the most commercially important species of the genus Coffea. It has been grown on the mountain terraces of southwestern Saudi Arabia for centuries. At present, the species is subject to increased genetic erosion due to the abandonment of many gardens by their owners and the increasingly dry climate. The current study was carried out to determine the genetic diversity of 56 local coffee accessions collected from the southern regions of Saudi Arabia using 30 sequence-related amplified polymorphism (SRAP) markers. Six SRAP markers showed polymorphism among the 56 accessions. A total of 1125 bands, with an average of 187.5, was produced from all six SRAP primers. The polymorphic information content (PIC) ranged from 74.8 to 97.7, with an average of 91.4 for all studied SRAP markers. The high polymorphism percentage seen in this study, along with the high number of alleles produced and the high PIC values of the primers used, demonstrate that the SRAP approach was an effective molecular technique for assessing genetic diversity in the studied populations. The structural analysis showed a sharp peak, with no ambiguity, demonstrating the highest delta K value at K = 3 and K = 6, and the coffee accessions could be grouped into three and six main populations, respectively. The PCoA, cluster analysis, and structural population analysis results suggest considerable genetic diversity among coffee populations growing on the southwestern mountain terraces of Saudi Arabia. The 56 accessions were segregated into five groups, mostly according to geographic distribution. The accessions from the southern districts of Jazan region mostly clustered in groups 2 and 4, while the accessions from the northern districts of Al-Baha and Assir regions formed separate groups. Based on these analyses, accessions KSA1R, KSA6R, KSA21, KSA25, KSA37, KSA38, KSA42, KSA59, KSA60, KSA62, and KSA63 were the most divergent. The genotypes should be conserved for use in coffee-breeding programs to improve the agronomic value of the crop, broaden the genetic base of C. arabica in Saudi Arabia and increase environmental resilience. Additional molecular and functional genomics studies are necessary to further elucidate how this germplasm has evolved and enhance the value of local Arabica coffee diversity in the Kingdom.