Polymorphism Information Content Research Articles

Assessing Genetic Variation among Strychnos spinosa Lam. Morphotypes Using Simple Sequence Repeat Markers.

Strychnos spinosa Lam., commonly known as green monkey orange, is a highly valued indigenous fruit tree in South Africa with potential for domestication and commercialization. However, no study has reported on the molecular diversity of Strychnos spinosa morphotypes. Therefore, this study aimed to determine genetic variation among 32 Strychnos spinosa morphotypes using simple sequence repeat (SSR) markers. Fourteen amplified SSR markers produced 159 alleles, with a mean of 5.68 per locus. The polymorphic information content (PIC) values ranged from 0.22 (Ssp_1) to 0.84 (Ssp_6). Morphotypes were clustered in a biplot based on their genetic distances. The dendrogram chiefly discriminated morphotypes according to variation of pericarp texture. The population structure had the highest delta value K = 3, thus the 32 morphotypes were divided into three subpopulations based on the Bayesian approach. The affinities produced by the population structure agreed with the genetic distance of closely related morphotypes. This study is the first to report on SSR marker development and their successful use for genetic diversity and population structure studies of Strychnos spinosa. It provides insights into the molecular characterisation of Strychnos spinosa. This can lead to breeding programs and crop improvement programs, particularly in varietal developmental programs, which can contribute to alleviating food security challenges.

Genetic diversity and structure of the coffee leaf rust fungus Hemileia vastatrix across different coffee management systems in Ethiopia.

Although coffee leaf rust (CLR), caused by Hemileia vastatrix, poses an increasing threat to coffee production in Ethiopia, little is known regarding its genetic diversity and structure and how these are affected by coffee management. Here, we used genetic fingerprinting based on sequence-related amplified polymorphism (SRAP) markers to genotype H. vastatrix samples from different coffee shrubs, across 40 sites, covering four coffee production systems (forest coffee, semi plantation coffee, home garden coffee, and plantation coffee) and different altitudes in Ethiopia. In total, 96 H. vastatrix samples were successfully genotyped with three primer combinations, producing a total of 79 scorable bands. We found 35.44% of amplified bands to be polymorphic, and the polymorphic information content (PIC) was 0.45, suggesting high genetic diversity among our CLR isolates. We also found significant isolation-by-distance across the samples investigated and detected significant differences in fungal genetic composition among plantation coffee and home garden coffee and a marginally significant difference among plantation coffee and forest coffee. Furthermore, we found a significant effect of altitude on CLR genetic composition in the forest coffee and plantation systems. Our results suggest that both spore dispersal and different selection pressures in the different coffee management systems are likely responsible for the observed high genetic diversity and genetic structure of CLR isolates in Ethiopia. When selecting Ethiopian coffee genotypes for crop improvement, it is important that these genotypes carry some resistance against CLR. Because our study shows large variation in genetic composition across relatively short geographical distances, a broad selection of rust isolates must be used for coffee resistance screening.

Genetic relationships and genome verification of Thai banana cultivars using Random Amplification of Polymorphic DNA (RAPD) markers

Abstract. Boonsrangsom T, Fuenghoi C, Premjet D, Suvittawat K, Ratanasut K, Sujipuli K. 2023. Genetic relationships and genome verification of Thai banana cultivars using Random Amplification of Polymorphic DNA (RAPD) markers. Biodiversitas 24: 3758-3765. Edible bananas and plantains, belonging to the family Musaceae, genus Musa, represent one of the most important fruit crops, with an annual production of more than 65 million tons worldwide. Bananas have several hybrid variations since they are descended from the two species Musa acuminata Colla (AA genome) and Musa balbisiana Colla (BB genome). Different morphological traits divide almost hybrid bananas into various genomic groupings. Banana genome categorization and identification, however, have always been challenging issues. This study aimed to assess the genetic relationships and verify the genomes of Thai banana cultivars using random amplification of polymorphic DNA (RAPD) markers. Using the 15 selected RAPD markers, 149 RAPD bands were found, with sizes ranging from 0.2 to 3.2 kb, and 88.6% were polymorphic. Polymorphic information content (PIC) values ranged from 0.18 to 0.42, averaging 0.30. Based on the Jaccard coefficient, the unweighted pair-group method arithmetic average (UPGMA) analysis showed that the banana samples had a similarity range of 0.27 to 1.00 with a mean of 0.56, demonstrating an abundance of viability across six banana genomes. The dendrogram generated from RAPD data revealed that all 18 Musa samples could be divided into two main groups (Group I and II). Three additional subgroups were created for each primary group (A, B, and C). The accurate identification and genetic data on the available genetic resources for bananas will be beneficial for breeding and conservation programs.

Genetic diversity analysis and fingerprints of Chrysanthemum×morifolium based on SSR molecular markers

The present study aims to explore the genetic diversity of germplasm resources of Chrysanthemum×morifolium (hereinafter, C.×morifolium) at the molecular level and to establish a fingerprint database of C.×morifolium varieties. We employed 12 pairs of primers with high levels of polymorphism, clear bands, and high degrees of reproducibility to analyze the SSR molecular markers and genetic diversity of 91 C.×morifolium materials and 14 chrysanthemum- related materials. With regard to constructing the fingerprints of the tested materials, we chose 9 pairs of core primers. The findings revealed that 12 primer pairs detected 104 alleles in 105 samples, ranging from 2 to 26. The average number of observed alleles (Na) per site was 9.25. The average number of effective alleles (Ne) per site was 2.745 6, with its range being 1.276 0 to 4.742 5. Shannon genetic diversity index (I) values ranged between 0.513 3 and 2.239 9 (M=1.209 0). Nei's gene diversity index (H) ranged between 0.216 3 and 0.789 1 (M=0.578 0). The observed heterozygosity (Ho) ranged between 0.223 3 and 0.895 2 (M=0.557 5). The expected heterozygosity (He) ranged between 0.217 4 and 0.793 3 (M=0.580 8). The polymorphism information content (PIC) ranged between 0.211 5 and 0.774 0 (M=0.532 9). The genetic similarity (GS) ranged between 0.228 5 and 1.000 0 (M=0.608 3). Cluster analysis revealed that when the genetic distance (GD) equals to 0.30, the tested materials can be classified into 2 groups. When the GD equals to 0.27, the first group can be divided into 6 subgroups; accordingly, 105 tested materials can be divided into 7 subgroups. The cophenetic correlation test was carried out based on the cluster analysis, and the corresponding results showed that the cluster map correlated with the genetic similarity coefficient (r=0.952 73). According to the results of Structure population analysis, we obtained the optimal population number, with the true number of populations (K) being 3 and the population being divided concerning Q≥0.5. Three subgroups, i.e., Q1, Q2 and Q3, included 34, 33 and 28 germplasms, respectively, and the remaining 10 germplasms were identified as the mixed population. During the experiment, 9 pairs of core primers were screened among the total of 12 for a complete differentiation regarding 105 tested materials, and the fingerprints of 91 C.×morifolium materials and 14 chrysanthemum-related materials were further constructed. Overall, there were significant genetic differences and rich genetic diversity among C.×morifolium materials, which would shed light on the garden application and variety selection fields of C.×morifolium. The fingerprint database of 105 C.×morifolium varieties and chrysanthemum-related species may provide technical support for future research regarding the identification and screening system of C.×morifolium varieties.

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.

Evaluation of soybean genotypes for cowpea mild mottle virus resistance through phenotypic reaction and genotypic analysis

AbstractCowpea mild mottle virus (CPMMV), a carlavirus nonpersistently transmitted by whiteflies, is an emerging and economically important virus of soybean. The present study aimed to identify soybean genotypes resistant to CPMMV through mechanical inoculation‐based phenotyping and marker trait association using gene‐specific and simple‐sequence repeat (SSR) markers. A set of 500 soybean genotypes was initially screened in the field for CPMMV infection, of which 288 genotypes showing resistance were evaluated further under controlled glasshouse conditions. Among these 288 genotypes, 43 (14.9%) were resistant, 59 (20.5%) were moderately resistant while the remaining 186 (64.6%) were susceptible. A set of five gene‐specific primer pairs, three of which targeted CPMMV R genes, was used for screening the 288 soybean genotypes; 96 genotypes from the 288 (based on disease score) were also genotyped with 24 SSR polymorphic markers for an association study. The mean values of major allele frequency and gene diversity were found to be 0.70 and 0.39, respectively. The polymorphism information content varied from 0.14 to 0.37. Population structure and cluster analysis revealed three subpopulations for the five gene‐specific markers and two distinct subpopulations for the 24 SSR markers, respectively. Neighbour joining‐based clustering analysis categorized the 288 genotypes into two major clusters. Three markers were found to be significantly associated with CPMMV resistance. The BARCSOYSSR0558 marker showed the highest phenotypic variance of 9.6%. The present study will help in identification of CPMMV R genes and breeding new resistant varieties through marker‐assisted selection.

Assessing the Genetic Variability of Sweet Chestnut Varieties from the Tuscan Apennine Mountains (Italy)

Castanea sativa Mill. is a valuable species with historical and economic importance in Europe, particularly in the Mediterranean area. In Italy, chestnut cultivation has been developed for centuries, leading to the recognition of more than 300 varieties. Nevertheless, a profusion of local names has been assigned by growers, causing the occurrence of synonyms and homonyms across the country. This research focused on genetic characterization and identification using 21 single sequence repeats (SSRs) for four chestnut varieties (i.e., Pastinese, Nerattino, Carpinese, and Rossola) commonly used for flour production in the Tuscan Apennine Mountains (Pistoia Province). A comprehensive number of 55 accessions identified by local growers as belonging to the four varieties were analyzed, in addition to a few “Marrone” accessions as outgroups. The 21 microsatellites were highly informative, detecting 98 alleles and displaying an average polymorphism information content (PIC) equal to 0.582. In addition, a considerable amount of genetic diversity was revealed, as shown by the heterozygosity levels (He = 0.634 and Ho = 0.475). The STRUCTURE analysis provided clear distinctions among the different varieties, splitting them into four separate groups. This result was also confirmed by UPGMA dendrogram and principal co-ordinates analysis (PCoA). However, one accession (Carp_5), previously identified as Carpinese, showed an allelic profile attributable to Pastinese, suggesting that farmers might have performed mislabeling or grafting propagation errors. Thus, our results confirm the use of SSRs to allocate the accessions of different varieties, uncovering possible synonyms and homonyms. Specifically, in the context of the Pistoiese mountain region, this tool can favor the traceability of processed products, such as flour, enhancing the quality and economic value of the local market. Lastly, our findings have revealed a considerable genetic variability within the Tuscan chestnut varieties whose preservation is mandatory to face climate change challenges through sustainable forest management practices.

Morphological and Molecular Diversity of Traditional Upland Rice (Oryza sativa L.) Varieties for Drought Tolerance at Seedling Stage

Drought is considered the major threat and most damaging abiotic stress for rice production.However, some traditional upland rice varieties (TURVs) may have some forms of tolerance to various stresses compared to new elite varieties. Two experiments were conducted. Experiment to determine the morpho-physiological response, phenotypic diversity, and similarity of 13 TURVs and 2 check varieties to mild drought stress at the seedling stage (Experiment 1 (E1) and to determine the molecular diversity and genetic similarity of the genotypes using selected simple sequence repeat (SSR) markers ((Experiment 2 (E2). In E1, all TURVs were highly tolerant under mild drought stress for a physiological response. Standardized Shannon-Weaver Diversity Index(H’) showed zero to low phenotypic diversity for qualitative traits. However, the phenotypic diversity of quantitative traits ranged from moderate to high for well-watered conditions, low to high for drought conditions, and moderate to high for re-watered conditions. Cluster analysis through Ward’s minimum variance based on pooled phenotypic data grouped the genotypes into three major clusters at 7 Euclidean distances. For E2, polymorphic information content values from 24 polymorphic SSR markers ranged from slightly informative (RM256) to highly informative (RM206), with a mean of 0.61. Nei’s gene diversity mean was 0.62 and values varied from 0.26 (RM256) to 0.86 (RM206). Based on Jaccard’s similarity coefficient, rice genotypes were grouped into nine minor clusters. The results indicate that TURVs harbor moderate to high allelic diversity and there are potential tolerance genes for mild drought stress that can be utilized in breeding programs for drought-tolerant rice varieties.

Genetic Diversity and Population Structure of African Sorghum (Sorghum bicolor L. Moench) Accessions Assessed through Single Nucleotide Polymorphisms Markers.

Assessing the genetic diversity and population structure of cultivated sorghum is important for heterotic grouping, breeding population development, marker-assisted cultivar development, and release. The objectives of the present study were to assess the genetic diversity and deduce the population structure of 200 sorghum accessions using diversity arrays technology (DArT)-derived single nucleotide polymorphism (SNP) markers. The expected heterozygosity values ranged from 0.10 to 0.50 with an average of 0.32, while the average observed heterozygosity (0.15) was relatively low, which is a typical value for autogamous crops species like sorghum. Moderate polymorphic information content (PIC) values were identified with a mean of 0.26, which indicates the informativeness of the chosen SNP markers. The population structure and cluster analyses revealed four main clusters with a high level of genetic diversity among the accessions studied. The variation within populations (41.5%) was significantly higher than that among populations (30.8%) and between samples within the structure (27.7%). The study identified distantly related sorghum accessions such as SAMSORG 48, KAURA RED GLUME; Gadam, AS 152; CSRO1, ICNSL2014-062; and YALAI, KAFI MORI. The accessions exhibited wide genetic diversity that will be useful in developing new gene pools and novel genotypes for West Africa sorghum breeding programs.

Analysis of genetic diversity and genetic structure of Qinchuan cattle conservation population using whole-genome resequencing.

In the conservation of livestock and poultry resources, population genetic diversity and genetic structure of the conservation population are important factors affecting the effectiveness of conservation. In this study, whole-genome resequencing technology was used to detect genomic variation in 100 Qinchuan cattle (30 bulls and 70 cows). By analyzing population genetic diversity, runs of homozygosity (ROH) distribution features, kinship relationships, and family structure, the conservation effectiveness of Qinchuan cattle was comprehensively evaluated. The results showed that a total of 20,968,017 high-quality SNPs were detected in 100 Qinchuan cattle, the average minimum allele frequency was 0.191±0.124, the average polymorphic information content was 0.279±0.131, and the average observed heterozygosity was 0.275±0.131, the average expected heterozygosity is 0.279±0.131, indicating that the genetic diversity of the Qinchuan cattle conservation population is relatively rich. The average identity by state (IBS) distance of the Qinchuan conservation population was 0.243±0.020, with a value of 0.242±0.021 for the bulls. The results of the kinship G-matrix were consistent with the results of the IBS distance matrix, both showing that some individuals in the conservation population had close kinship. A total of 8258 genomic ROH were detected in 100 Qinchuan cattle, with a total length of 9.64 GB. The average length of ROH fragments was 1.167±1.203 Mb, 69.35% of the ROH were short ROH with a length of 0.5~1 Mb, and the average total length of ROH per individual was 96.40 Mb. The average inbreeding coefficient based on ROH was 0.039±0.039, with a value of 0.044±0.035 for the bulls, indicating that some bulls had a certain degree of inbreeding accumulation. The results of the phylogenetic tree combined with kinship analysis showed that the individuals in the Qinchuan cattle conservation population could be divided into eight families, including seven families with bulls and one family without bulls. This study demonstrated that the genetic diversity of the Qinchuan conservation population is relatively rich, with no significant inbreeding accumulation, but there is a risk of inbreeding among some individuals. Therefore, it is necessary to strengthen selection and mating to ensure the sustainable development of Qinchuan cattle resources.

Population Structure and Genetic Diversity of Rice (Oryza sativa L.) Germplasm from the Democratic Republic of Congo (DRC) Using DArTseq-Derived Single Nucleotide Polymorphism (SNP)

Understanding the genetic diversity and population structure of rice is crucial for breeding programs, conservation efforts, and the development of sustainable agricultural practices. This study aimed to assess the genetic diversity and population structure of 94 rice (Oryza sativa L.) genotypes from the Democratic Republic of Congo using a set of 8389 high-quality DArTseq-based single nucleotide polymorphism (SNP) markers. The average polymorphic information content (PIC) of the markers was 0.25. About 42.4% of the SNPs had a PIC value between 0.25 and 0.5, which were moderately informative. The ADMIXTURE program was used for structure analysis, which revealed five sub-populations (K = 5), with admixtures. In principal component analysis (PCA), the first three principal components accounted for 36.3% of the total variation. Analysis of molecular variance revealed significant variation between sub-populations (36.09%) and within genotypes (34.04%). The low overall number of migrants (Nm = 0.23) and high fixation index (Fst = 0.52) indicated limited gene flow and significant differentiation between the sub-populations. Observed heterozygosity (Ho = 0.08) was lower than expected heterozygosity (He = 0.14) because of the high inbreeding (Fis = 0.52) nature of rice. A high average Euclidean genetic distance (0.87) revealed the existence of genetic diversity among the 94 genotypes. The significant genetic diversity among the evaluated rice genotypes can be further explored to obtain potentially desirable genes for rice improvement.

Genetic Diversity of Cowpea Parental Lines Assembled for Breeding in Uganda

Cowpea is a grain legume that is commonly grown in Sub-Saharan Africa (SSA) for a variety of uses including food, feed, and soil fertility enhancement through nitrogen fixation. Genetic improvement in cowpea can be accomplished by creating breeding populations from high-performing but genetically diverse parental lines. The goal of this study was to use single nucleotide polymorphic markers (SNPs) to estimate genetic diversity and population stratification among cowpea parental genotypes from five countries. A total of 2,746 SNP markers were used to analyze 62 cowpea genotypes that differed in key characteristics. Only 55% of the SNP markers were polymorphic across the 62 genotypes and met the selection requirement. The gene diversity varied from 0.52 to 0.57 with an average of 0.55, while the polymorphic information content ranged from 0.46 to 0.51 with an average of 0.49, showing that the cowpea parental lines are genetically diverse. The analysis of molecular variance (AMOVA) found high significant (p < 0.001) variation among genotypes and non-significant variation across genotypes depending on country of origin. The cluster analysis for the 62 genotypes using the unweighted pair group procedure with arithmetic mean approach divided the genotypes into two distinct groups. Genotypes from the same geographical regions tended to cluster together and there was a significant level of genetic variation among genotypes from Uganda and Nigeria. The SNP markers utilized in this study were effective at distinguishing genotypes and may be used to estimate the diversity in the cowpea core collection. The genetic diversity in this collection is important for future development of improved varieties of cowpea in Uganda.

Evaluation of the Genetic Diversity of Greek Garlic (Allium sativum L.) Accessions Using DNA Markers and Association with Phenotypic and Chemical Variation

The genetic diversity of 27 garlic accessions (local varieties/landraces) was investigated using five simple sequence repeat (SSR) and six inter-simple sequence repeat (ISSR) markers. The SSR genotyping revealed 26 different alleles, while 84 polymorphic bands were produced using the ISSR markers. The average polymorphism information content (PIC) was 0.471 and 0.465 for the SSR and the ISSR markers, respectively. Compared to ISSRs, SSR markers revealed a higher level of redundancy, indicating potential duplicates among the accessions. The analysis of molecular variance (AMOVA) based on both marker systems showed that most of the molecular diversity was attributable to differences within accessions, rather than among them. UPGMA, STRUCTURE, and Principal Coordinate Analysis (PCoA) based on the SSRs produced similar clustering results, although not in agreement with those produced by the ISSR markers. Fisher’s exact tests and multinomial logistic regression analyses identified significant associations between the chemical compounds, the morphological traits of the bulb, and specific SSR and ISSR loci. Our results provide a molecular basis for understanding the genetic diversity of Greek garlic landraces, which could be useful for the conservation and sustainable management of this crop. Potential markers fostering the selection of genotypes in garlic breeding have also been revealed.

Genetic diversity and population structure of indigenous guinea fowl (Numida meleagris) in Benin using simple sequence repeat markers.

This study aimed to assess the genetic diversity and population structure of 12 guinea fowl phenotypes from three climatic zones (Guinean, Sudano-Guinean and Sudanian) of Benin. A total of 96 adult guinea fowl, aged at least 6months, were selected for blood sampling. Fragment analysis was carried out using 17 polymorphic microsatellite markers. The informative marker combinations revealed a total of 83alleles across all loci, with an average of 5alleles per locus and a mean polymorphic information content (PIC) of0.793. This study showed an observed heterozygosity of0.492. The inbreeding coefficient values ranged from0.086 in white phenotype to0.226 in cinnamon, showing a deficit of heterozygotes, suggesting a moderate inbreeding level. A relatively low population differentiation was observed, with a mean fixation index(Fst) value of0.033. The short genetic distances between phenotypes, unlike the strong genetic identities, revealed high genetic proximity between the 12 phenotypes of indigenous guinea fowl in Benin. These data indicate the existence of a single indigenous guinea fowl population with high intra-population genetic diversity with respect to climatic zones or phenotypes. This study will help in the selection of parental breeding stock for genetic improvement programs, as well as in the conservation for biodiversity maintenance and sustainable use of the indigenous guinea fowl in the study zones in Benin.

Comparative analysis of nine Tilletia indica genomes for the development of novel microsatellite markers for genetic diversity and population structure analysis.

Karnal bunt (KB; Tilletia indica) is the prime quarantine concern for quality wheat production throughout the world. The most effective approach to dealing with this biotic stress is to breed KB-resistant wheat varieties, which warrants a better understanding of T. indica genome architecture. In India, the North Western Plain Zone is the prime hot spot for KB disease, but only limited efforts have been made to decipher T. indica diversity at the genomic level. Microsatellites offer a powerful and robust typing system for the characterization and genetic diversity assessment of plant pathogens. At present, inadequate information is available with respect to the development of genome-derived markers for revealing genetic variability in T. indica populations. In current research, nine complete genome sequences of T. indica (PSWKBGH_1, PSWKBGH_2, PSWKBGD_1_3, RAKB_UP_1, TiK_1, Tik, DAOMC236408, DAOMC236414, and DAOMC236416) that exist in the public domain were explored to know the dynamic distribution of microsatellites. Comparative genome analysis revealed a high level of relative abundance and relative density of microsatellites in the PSWKBGH_1 genome in contrast to other genomes. No significant correlation between microsatellite distribution for GC content and genome size was established. All the genomes showed the dominance of tri-nucleotide motifs, followed by mono-, di-, tetra-, hexa-, and penta-nucleotide motifs. Out of 50 tested markers, 36 showed successful amplification in T. indica isolates and produced 52 different alleles. A PCR assay along with analysis of the polymorphic information content (PIC) revealed 10 markers as neutral and polymorphic loci (PIC 0.37). The identified polymorphic SSR loci grouped a geographically distinct T. indica population of 50 isolates representing seven Indian regions (Jammu, Himachal Pradesh, Punjab, Haryana, Uttarakhand, Uttar Pradesh, and Rajasthan) into four distinct clusters. The results of the analysis of molecular variance identified 94% genetic variation within the population and 6% among the population. Structure analysis also confirmed the existence of four genetically diverse groups containing admixtures of T. indica isolates across populations. In nutshell, the current study was successful in identifying novel, neutral and polymorphic microsatellite markers that will be valuable in offering deep insight into the evolutionary relationship and dynamics of the T. indica population for devising effective KB management strategies in wheat.

Assessment of Genetic Diversity in the White-Colored Variants of Spray-Type Chrysanthemum Cultivars Using SSR Markers

Chrysanthemums represent the second most important cut flower after rose on the global commercial market. The phenomenal importance and global popularity of chrysanthemums have attracted breeders’ attention, resulting in the release of vast numbers of cultivars. Identifying these cultivars is crucial to protecting breeders’ intellectual property rights and improving the efficiency of breeding. Distinguishing chrysanthemum genotypes based on their morphological characteristics is challenging as they vary highly within this group, hence requiring the use of efficient molecular markers. In this study, we evaluated the genetic diversity of 57 spray-type chrysanthemum cultivars bearing white, ivory, and cream-colored flowers. A total of six loci were evaluated regarding their polymorphism efficiency across the tested cultivars. Allele numbers ranged from 2 to 6, with a mean of 3.5 alleles per locus. The average polymorphism information content (PIC) was 0.53 for six SSR markers. Cluster analysis of genetic relationships using the UPGMA method showed a genetic distance of 0.31 to 1.00, and the 57 white variants of chrysanthemum cultivars were characterized using the tested SSR markers. However, two sets of cultivars, namely, Pure Angel–Neba and Ladost–White wing, exhibited total genetic similarity and hence could not be discriminated. These results provide efficient SSR markers that can be used to identify chrysanthemum cultivars (and assess their genetic relationships) that cannot be discriminated based on phenotype.

De Novo Mining and Validating Novel Microsatellite Markers to Assess Genetic Diversity in Maruca vitrata (F.), a Legume Pod Borer.

Maruca vitrata (Fabricius) is an invasive insect pest capable of causing enormous economic losses to a broad spectrum of leguminous crops. Microsatellites are valuable molecular markers for population genetic studies; however, an inadequate number of M. vitrata microsatellite loci are available to carry out population association studies. Thus, we utilized this insect's public domain databases for mining expressed sequence tags (EST)-derived microsatellite markers. In total, 234 microsatellite markers were identified from 10053 unigenes. We discovered that trinucleotide repeats were the most predominant microsatellite motifs (61.53%), followed by dinucleotide repeats (23.50%) and tetranucleotide repeats (14.95%). Based on the analysis, twenty-five markers were selected for validation in M. vitrata populations collected from various regions of India. The number of alleles (Na), observed heterozygosity (Ho), and expected heterozygosity (He) ranged from 2 to 5; 0.00 to 0.80; and 0.10 to 0.69, respectively. The polymorphic loci showed polymorphism information content (PIC), ranging from 0.09 to 0.72. Based on the genetic distance matrix, the unrooted neighbor-joining dendrogram differentiated the selected populations into two discrete groups. The SSR markers developed and validated in this study will be helpful in population-level investigations of M. vitrata to understand the gene flow, demography, dispersal patterns, biotype differentiation, and host dynamics.

Characterisation of bell pepper (Capsicum annuum L. var. grossum Sendt.) accessions for genetic diversity and population structure based on agro-morphological and microsatellite markers

Bell pepper (Capsicum annuum L. var. grossum Sendt.) is a leading Solanaceous vegetable crop across the world with immense therapeutic and export potential. Such wide cultivation exposes the crop to various biotic and abiotic stresses. Genetic diversity is one of the major prerequisites for any crop breeding programme. Hence, in this study, an attempt was made to evaluate the genetic diversity and population structure of 24 bell pepper genotypes using 25 agro-morphological [9 distinctness, uniformity and stability (DUS) descriptors; 13 quantitative traits; and 3 quality traits] and 82 microsatellite or simple sequence repeat (SSR) markers. Eight DUS descriptors showed polymorphism with maximum morphological variation (H = 1.29) for fruit shape in longitudinal section. Multivariate analysis using Mahalanobis D2 statistic distinguished the genotypes into 11 clusters. With a mean of 4.08 alleles per locus, a total of 151 alleles were generated from 37 polymorphic SSR primers. The polymorphic information content (PIC) ranged from 0.14 (CAMS 451) to 0.76 (CA 525390) with an average of 0.54. The genotypes were divided into two major clusters based on cluster analysis using the unweighted pair group method with arithmetic mean (UPGMA) algorithm, whereas the cluster tree using the neighbour-joining (NJ) method identified three major groups. The population was classified into three groups based on structure analysis. On comparing agro-morphological and molecular clusters, 11 genotypes viz., BWT-39, BWT-39-BR, BWT-39-DR, BWT-29, BWT-3Y, BWT-3Y-4L, Kandaghat Selection, BWT-48-AC, BWT-49-AC, EC-464107, and EC-464115, were found common. These genotypes also exhibited considerable resistance to bacterial wilt disease. Mantel's test showed a moderate positive correlation between agro-morphological (DUS descriptors) and molecular data. Therefore, a combined approach of agro-morphological and molecular evaluation can facilitate the selection of diverse parental lines followed by subsequent hybridisation between them to introduce new traits in bell pepper.

Genetic diversity and conservation of Siberian apricot (Prunus sibirica L.) based on microsatellite markers

Siberian apricot (Prunus sibirica L.) is a woody tree species of ecological, economic, and social importance. To evaluate the genetic diversity, differentiation, and structure of P. sibirica, we analyzed 176 individuals from 10 natural populations using 14 microsatellite markers. These markers generated 194 alleles in total. The mean number of alleles (13.8571) was higher than the mean number of effective alleles (6.4822). The average expected heterozygosity (0.8292) was higher than the average observed heterozygosity (0.3178). Shannon information index and polymorphism information content were separately 2.0610 and 0.8093, demonstrating the rich genetic diversity of P. sibirica. Analysis of molecular variance revealed that 85% of the genetic variation occurred within populations, with only 15% among them. The genetic differentiation coefficient and gene flow were separately 0.151 and 1.401, indicating a high degree of genetic differentiation. Clustering results showed that a genetic distance coefficient of 0.6 divided the 10 natural populations into two subgroups (subgroups A and B). STRUCTURE and principal coordinate analysis divided the 176 individuals into two subgroups (clusters 1 and 2). Mantel tests revealed that genetic distance was correlated with geographical distance and elevation differences. These findings can contribute to the effective conservation and management of P. sibirica resources.

Genetic Analysis of the Cultivars of Ping’ou Hybrid Hazelnut (C. heterophylla Fisch. × C. avellana L.) in China Based on SSR Markers

Ping’ou hybrid hazelnut is one of the most profitable tree nuts in China, but economically important cultivars must first be genetically validated to meet industrial demand. Traditional approaches used for cultivar identification are mainly trait-based and unreliable. Previous approaches at the DNA level, focusing on the identification of species or/and varieties that originated in China, were not used widely in hybrid hazelnut because there was no proper standard sample. In this research, a multiplexed fingerprinting test was conducted to allow for hazelnut cultivar identification using SSR markers derived from European hazelnut. Twenty-seven SSR markers were used to fingerprint 57 genetically unique Ping’ou hybrid hazelnut and related wild species. All markers showed a high level of polymorphism, as indicated by mean values for observed heterozygosity (Ho = 0.84), expected heterozygosity (He = 0.80), and polymorphism information content (PIC = 0.78). A total of 301 alleles were detected, and the number of effective alleles varied from 6 for KG817 and GB818 to 18 for B654, with an average of 11.2 alleles per locus. Moreover, the Shannon’s information index (I) ranged from 1.293 for BR215 to 2.385 for B654, with an average of 1.908. The neighbor-joining tree, principal coordinate analysis, and Bayesian analysis revealed clear separation between hybrid cultivars and wild forms (Cluster/group I), as well as the differentiation within hybrid genotypes (Clusters/groups II and III). Additionally, the NJ dendrogram demonstrated a further split within Clusters/group III (III a and III b). Altogether, with the comparable SSR information of the European hazelnut cultivar ‘Barcelona’, the newly developed marker sets can assist in the germplasm identification of hazelnut cultivars and reproductive materials. Importantly, these combined SSR loci can be applied to characterize the genetic relationships and population structures among wild genotypes and hybrid cultivars, which will then provide information to guide hazelnut breeding based on their genetic background.