Genetic diversity assessment of hydrogen cyanide, total carotenoid content, and dry matter content in biofortified cassava using trait-linked SNP markers

Radish (Raphanus sativus L.) is a vegetable crop with economic value and ecological significance in the genus Radish, family Brassicaceae. In recent years, developed countries have attached great importance to the collection and conservation of radish germplasm resources and their research and utilization, but the lack of population genetic information and molecular markers has hindered the development of the genetic breeding of radish. In this study, we integrated the radish genomic data published in databases for the development of single-nucleotide polymorphism (SNP) markers, and obtained a dataset of 308 high-quality SNPs under strict selection criteria. With the support of Kompetitive Allele-Specific PCR (KASP) technology, we screened a set of 32 candidate core SNP marker sets to analyse the genetic diversity of the collected 356 radish varieties. The results showed that the mean values of polymorphism information content (PIC), minor allele frequency (MAF), gene diversity and heterozygosity of the 32 candidate core SNP markers were 0.32, 0.30, 0.40 and 0.25, respectively. Population structural analysis, principal component analysis and genetic evolutionary tree analysis indicated that the 356 radish materials were best classified into two taxa, and that the two taxa of the material were closely genetically exchanged. Finally, on the basis of 32 candidate core SNP markers we calculated 15 core markers using a computer algorithm to construct a fingerprint map of 356 radish varieties. Furthermore, we constructed a core germplasm population consisting of 71 radish materials using 32 candidate core markers. In this study, we developed SNP markers for radish cultivar identification and genetic diversity analysis, and constructed DNA fingerprints, providing a basis for the identification of radish germplasm resources and molecular marker-assisted breeding as well as genetic research.

Genetic approaches have proved valuable to the study and conservation of endangered populations, especially for monitoring programs, and there is potential for further developments in this direction by extending analyses to the genomic level. We assembled the genome of the wolverine (Gulo gulo), a mustelid that in Scandinavia has recently recovered from a significant population decline, and obtained a 2.42Gb draft sequence representing >85% of the genome and including >21,000protein-coding genes. We then performed whole-genome resequencing of 10Scandinavian wolverines for population genomic and demographic analyses. Genetic diversity was among the lowest detected in a red-listed population (mean genome-wide nucleotide diversity of 0.05%). Results of the demographic analyses indicated a long-term decline of the effective population size (Ne ) from 10,000well before the last glaciation to <500 after this period. Current Ne appeared even lower. The genome-wide FIS level was 0.089 (possibly signaling inbreeding), but this effect was not observed when analyzing a set of highly variable SNP markers, illustrating that such markers can give a biased picture of the overall character of genetic diversity. We found significant population structure, which has implications for population connectivity and conservation. We used an integrated microfluidic circuit chip technology to develop an SNP-array consisting of 96 highly informative markers that, together with a multiplex pre-amplification step, was successfully applied to low-quality DNA from scat samples. Our findings will inform management, conservation, and genetic monitoring of wolverines and serve as a genomic roadmap that can be applied to other endangered species. The approach used here can be generally utilized in other systems, but we acknowledge the trade-off between investing in genomic resources and direct conservation actions.

Characterization of genetic diversity, population structure, and linkage disequilibrium is a prerequisite for proper management of breeding programs and conservation of genetic resources. In this study, 186 chickpea genotypes, including advanced “Kabuli” breeding lines and Iranian landrace “Desi” chickpea genotypes, were genotyped using DArTseq-Based single nucleotide polymorphism (SNP) markers. Out of 3339 SNPs, 1152 markers with known chromosomal position were selected for genome diversity analysis. The number of mapped SNP markers varied from 52 (LG8) to 378 (LG4), with an average of 144 SNPs per linkage group. The chromosome size that was covered by SNPs varied from 16,236.36 kbp (LG8) to 67,923.99 kbp (LG5), while LG4 showed a higher number of SNPs, with an average of 6.56 SNPs per Mbp. Polymorphism information content (PIC) value of SNP markers ranged from 0.05 to 0.50, with an average of 0.32, while the markers on LG4, LG6, and LG8 showed higher mean PIC value than average. Unweighted neighbor joining cluster analysis and Bayesian-based model population structure grouped chickpea genotypes into four distinct clusters. Principal component analysis (PCoA) and discriminant analysis of principal component (DAPC) results were consistent with that of the cluster and population structure analysis. Linkage disequilibrium (LD) was extensive and LD decay in chickpea germplasm was relatively low. A few markers showed r2 ≥ 0.8, while 2961 pairs of markers showed complete LD (r2 = 1), and a huge LD block was observed on LG4. High genetic diversity and low kinship value between pairs of genotypes suggest the presence of a high genetic diversity among the studied chickpea genotypes. This study also demonstrates the efficiency of DArTseq-based SNP genotyping for large-scale genome analysis in chickpea. The genotypic markers provided in this study are useful for various association mapping studies when combined with phenotypic data of different traits, such as seed yield, abiotic, and biotic stresses, and therefore can be efficiently used in breeding programs to improve chickpea.

This study seeks to determine the genetic diversity among cassava landraces using single nucleotide polymorphic (SNP) markers. One hundred and five cassava landraces were assayed with 195 SNP markers. Major allele frequency varied from 0.500 to 0.942 with an average of 0.728. Average gene diversity, heterozygosity and polymorphic information content (PIC) were 0.359, 0.314 and 0.286 respectively. These values were generally high considering the bi-allelic nature of SNPs, hence the cassava landraces studied showed moderate to high genetic diversity. This suggests availability of unique and useful alleles that could be exploited for breeding purposes. Inclusion of these landraces in our crop improvement activities will enhance the development of farmer preferred cassava varieties. SNP markers used for the study were highly informative, polymorphic and revealed good estimates of genetic diversity among the landraces. Higher level of genetic variation was observed within population based on analysis of molecular variance (AMOVA). Principal component analysis (PCA) and cluster analysis also grouped landraces into three distinct clusters; however, they did not group in accordance to geographical origin. This could be due to high frequency of germplasm exchange between farmers and subsequent change of the name of the same cultivar. Results from this study may contribute significantly to cassava breeding and germplasm conservation programs. Key words: Genetic diversity, single nucleotide polymorphisms (SNPs), polymorphic information content (PIC), polymorphic, alleles, heterozygosity, germplasm.

Cashew (Anacardium occidentale L.) is an important tree grown worldwide for its edible fruits, nuts and other products of industrial applications. The ecologically sensitive cashew-growing region in coastal Kenya is significantly affected by rising temperatures, droughts, floods, and shifting rainfall patterns. These changes adversely impact cashew growth by altering flowering patterns, increasing pests and diseases, and causing postharvest losses, which ultimately result in reduced yields and tree mortality. This is exacerbated by the long juvenile phase, high heterozygosity, lack of trait correlations, large mature plant size, and inadequate genomic resources. For the first time, the Diversity Array Technology (DArT) technology was employed to identify DArT (silicoDArT) and single nucleotide polymorphisms (SNPs) markers for genomic understanding of cashew in Kenya. Cashew leaf samples were collected in Kwale, Kilifi and Lamu counties along coastal Kenya followed by DNA extraction. The reduced libraries were sequenced using Hiseq 2500 Illumina sequencer, and the SNPs called using DarTsoft14. A total of 27,495 silicoDArT and 17,008 SNP markers were reported, of which 1340 silicoDArT and 824 SNP markers were used for analyses after screening, with > 80% call rate, > 95% reproducibility, polymorphism information content (PIC ≥ 0.25) and one ratio (>0.25). The silicoDArT and SNP markers had mean PIC values ranging from 0.02-0.50 and 0.0-0.5, with an allelic richness ranging from 1.992 to 1.994 for silicoDArT and 1.862 to 1.889 for SNP markers. The observed heterozygosity and expected values ranged from 0.50-0.55 and 0.34-0.37, and 0.56-0.57 and 0.33 for both silicoDArT and SNP markers respectively. Understanding cashew genomics through the application of SilicoDArT and SNP markers is crucial for advancing cashew genomic breeding programs aimed at improving yield and nut quality, and enhancing resistance or tolerance to biotic and abiotic stresses. Our study presents an overview of the genetic diversity of cashew landraces in Kenya and demonstrates that DArT systems are a reliable tool for advancing genomic research in cashew breeding.

Understanding germplasm’s genetic diversity is essential for developing new and improved cultivars with stable yields under diverse environments. The objective of this study was to determine the genetic diversity and population structure of 128 maize inbred lines sourced from the International Institute of Tropical Agriculture (IITA), the International Maize and Wheat Improvement Centre (CIMMYT), and the University of KwaZulu-Natal (UKZN) using 11,450 informative single nucleotide polymorphism (SNP) markers. The inbred lines revealed highly significant (p < 0.001) levels of variability for the key phenotypic traits. The SNP markers had a mean gene diversity (GD) and polymorphic information content (PIC) of 0.40 and 0.31, respectively, indicating the existence of substantial genetic variation across the germplasm panel. The model-based population structure analysis identified three subpopulations (K = 3) among the inbred lines. This corroborated the phylogenetic analysis using phenotypic traits and molecular markers which classified the inbred lines into three groups. The findings of this study identified considerable genetic diversity for the selection of inbred lines with favourable alleles for multiple traits and could be useful to initiate marker-assisted selection (MAS) to identify significant loci associated with agronomic performance and multiple-stress tolerance.

Germplasm resources embody the genetic diversity of plants and form the foundation for breeding and the ongoing improvement of elite cultivars. The establishment of germplasm banks, along with their systematic evaluation, constitutes a critical step toward the conservation, sustainable use, and innovative utilization of these resources. Liriodendron, a rare and endangered tree genus with species distributed in both East Asia and North America, holds considerable ecological, ornamental, and economic significance. However, a standardized evaluation system for Liriodendron germplasm remains unavailable. In this study, 297 Liriodendron germplasm accessions were comprehensively evaluated using 34 phenotypic traits and whole-genome resequencing data. Substantial variation was observed in most phenotypic traits, with significant correlations identified among several characteristics. Cluster analysis based on phenotypic data grouped the accessions into three distinct clusters, each exhibiting unique distribution patterns. This classification was further supported by principal component analysis (PCA), which effectively captured the underlying variation among accessions. These phenotypic groupings demonstrated high consistency with subsequent population structure analysis based on SNP markers (K = 3). Notably, several key traits exhibited significant divergence (p < 0.05) among distinct genetic clusters, thereby validating the coordinated association between phenotypic variation and molecular markers. Genetic diversity and population structure were assessed using 4204 high-quality single-nucleotide polymorphism (SNP) markers obtained through stringent filtering. The results indicated that the Liriodendron sino-americanum displayed the highest genetic diversity, with an expected heterozygosity (He) of 0.18 and a polymorphic information content (PIC) of 0.14. In addition, both hierarchical clustering and PCA revealed clear population differentiation among the accessions. Association analysis between three phenotypic traits (DBH, annual height increment, and branch number) and SNPs identified 25 highly significant SNP loci (p < 0.01). Of particular interest, the branch number-associated locus SNP_17_69375264 (p = 1.03 × 10−5) demonstrated the strongest association, highlighting distinct genetic regulation patterns among different growth traits. A minimal set of 13 core SNP markers was subsequently used to construct unique DNA fingerprints for all 297 accessions. In conclusion, this study systematically characterized phenotypic traits in Liriodendron, identified high-quality and core SNPs, and established correlations between key phenotypic and molecular markers. These achievements enabled differential analysis and genetic diversity assessment of Liriodendron germplasm, along with the construction of DNA fingerprint profiles. The results provide crucial theoretical basis and technical support for germplasm conservation, accurate identification, and utilization of Liriodendron resources, while offering significant practical value for variety selection, reproduction and commercial applications of this species.

Finger millet is an important cereal crop in eastern Africa and southern India with excellent grain storage quality and unique ability to thrive in extreme environmental conditions. Since negligible attention has been paid to improving this crop to date, the current study used Next Generation Sequencing (NGS) technologies to develop both Simple Sequence Repeat (SSR) and Single Nucleotide Polymorphism (SNP) markers. Genomic DNA from cultivated finger millet genotypes KNE755 and KNE796 was sequenced using both Roche 454 and Illumina technologies. Non-organelle sequencing reads were assembled into 207 Mbp representing approximately 13% of the finger millet genome. We identified 10,327 SSRs and 23,285 non-homeologous SNPs and tested 101 of each for polymorphism across a diverse set of wild and cultivated finger millet germplasm. For the 49 polymorphic SSRs, the mean polymorphism information content (PIC) was 0.42, ranging from 0.16 to 0.77. We also validated 92 SNP markers, 80 of which were polymorphic with a mean PIC of 0.29 across 30 wild and 59 cultivated accessions. Seventy-six of the 80 SNPs were polymorphic across 30 wild germplasm with a mean PIC of 0.30 while only 22 of the SNP markers showed polymorphism among the 59 cultivated accessions with an average PIC value of 0.15. Genetic diversity analysis using the polymorphic SNP markers revealed two major clusters; one of wild and another of cultivated accessions. Detailed STRUCTURE analysis confirmed this grouping pattern and further revealed 2 sub-populations within wild E. coracana subsp. africana. Both STRUCTURE and genetic diversity analysis assisted with the correct identification of the new germplasm collections. These polymorphic SSR and SNP markers are a significant addition to the existing 82 published SSRs, especially with regard to the previously reported low polymorphism levels in finger millet. Our results also reveal an unexploited finger millet genetic resource that can be included in the regional breeding programs in order to efficiently optimize productivity.

Simple sequence repeat (SSR) and single nucleotide polymorphism (SNP) markers are amongst the most common markers of choice for studies of diversity and relationships in horticultural species. We have used 11 SSR and 35 SNP markers derived from transcriptome sequencing projects to fingerprint 48 accessions of a collection of brinjal (Solanum melongena), gboma (S. macrocarpon) and scarlet (S. aethiopicum) eggplant complexes, which also include their respective wild relatives S. incanum, S. dasyphyllum and S. anguivi. All SSR and SNP markers were polymorphic and 34 and 36 different genetic fingerprints were obtained with SSRs and SNPs, respectively. When combining both markers all accessions but two had different genetic profiles. Although on average SSRs were more informative than SNPs, with a higher number of alleles, genotypes and polymorphic information content (PIC), and expected heterozygosity (H e ) values, SNPs have proved highly informative in our materials. Low observed heterozygosity (H o ) and high fixation index (f) values confirm the high degree of homozygosity of eggplants. Genetic identities within groups of each complex were higher than with groups of other complexes, although differences in the ranks of genetic identity values among groups were observed between SSR and SNP markers. For low and intermediate values of pair-wise SNP genetic distances, a moderate correlation between SSR and SNP genetic distances was observed (r2 = 0.592), but for high SNP genetic distances the correlation was low (r2 = 0.080). The differences among markers resulted in different phenogram topologies, with a different eggplant complex being basal (gboma eggplant for SSRs and brinjal eggplant for SNPs) to the two others. Overall the results reveal that both types of markers are complementary for eggplant fingerprinting and that interpretation of relationships among groups may be greatly affected by the type of marker used.

Information about the genetic diversity and population structure in elite breeding material is of fundamental importance for the improvement of crops. The objectives of our study were to (a) examine the population structure and the genetic diversity in elite maize germplasm based on simple sequence repeat (SSR) markers, (b) compare these results with those obtained from single nucleotide polymorphism (SNP) markers, and (c) compare the coancestry coefficient calculated from pedigree records with genetic distance estimates calculated from SSR and SNP markers. Our study was based on 1,537 elite maize inbred lines genotyped with 359 SSR and 8,244 SNP markers. The average number of alleles per locus, of group specific alleles, and the gene diversity (D) were higher for SSRs than for SNPs. Modified Roger’s distance (MRD) estimates and membership probabilities of the STRUCTURE matrices were higher for SSR than for SNP markers but the germplasm organization in four heterotic pools was consistent with STRUCTURE results based on SSRs and SNPs. MRD estimates calculated for the two marker systems were highly correlated (0.87). Our results suggested that the same conclusions regarding the structure and the diversity of heterotic pools could be drawn from both markers types. Furthermore, although our results suggested that the ratio of the number of SSRs and SNPs required to obtain MRD or D estimates with similar precision is not constant across the various precision levels, we propose that between 7 and 11 times more SNPs than SSRs should be used for analyzing population structure and genetic diversity.Electronic supplementary materialThe online version of this article (doi:10.1007/s00122-009-1256-2) contains supplementary material, which is available to authorized users.

BackgroundAssessment and effective utilization of genetic diversity in breeding programs is crucial for sustainable genetic improvement and rapid adaptation to changing breeding objectives. During the past two decades, the commercialization of the early and extra-early maturing cultivars has contributed to rapid expansion of maize into different agro-ecologies of sub-Saharan Africa (SSA) where maize has become an important component of the agricultural economy and played a vital role in food and nutritional security. The present study aimed at understanding the population structure and genetic variability among 439 early and extra-early maize inbred lines developed from three narrow-based and twenty-seven broad-based populations by the International Iinstitute of Tropical Agriculture Maize Improvement Program (IITA-MIP). These inbreds were genotyped using 9642 DArTseq-based single nucleotide polymorphism (SNP) markers distributed uniformly throughout the maize genome.ResultsAbout 40.8% SNP markers were found highly informative and exhibited polymorphic information content (PIC) greater than 0.25. The minor allele frequency and PIC ranged from 0.015 to 0.500 and 0.029 to 0.375, respectively. The STRUCTURE, neighbour-joining phylogenetic tree and principal coordinate analysis (PCoA) grouped the inbred lines into four major classes generally consistent with the selection history, ancestry and kernel colour of the inbreds but indicated a complex pattern of the genetic structure. The pattern of grouping of the lines based on the STRUCTURE analysis was in concordance with the results of the PCoA and suggested greater number of sub-populations (K = 10). Generally, the classification of the inbred lines into heterotic groups based on SNP markers was reasonably reliable and in agreement with defined heterotic groups of previously identified testers based on combining ability studies.ConclusionsComplete understanding of potential heterotic groups would be difficult to portray by depending solely on molecular markers. Therefore, planned crosses involving representative testers from opposing heterotic groups would be required to refine the existing heterotic groups. It is anticipated that the present set of inbreds could contribute new beneficial alleles for population improvement, development of hybrids and lines with potential to strengthen future breeding programs. Results of this study would help breeders in formulating breeding strategies for genetic enhancement and sustainable maize production in SSA.

The demand for tomato is increasing day by day mostly because of the increased per capita fresh fruit consumption. Nonetheless as a perishable fruit crop, it has relatively short life after ripening thus experiences remarkable post-harvest losses. The study was to select true F1 hybrids using SNPs for the development of inbred lines with long shelf life character through marker assisted backcrossing. Nine out of the one hundred and forty SNP markers failed, 93 were uninformative and 38 were polymorphic and spread across the 12 chromosomes. The major allele frequency, gene diversity, heterozygosity and Polymorphic Information Content (PIC) for each locus were calculated for SNP markers using Power Marker 3.5. The mean value of the major allele frequency was 0.673, ranging between 0.529 and 0.794. The average gene diversity and Heterozygosity values were 0.419 and 0.125 respectively. The PIC ranged from 0.273 (Sly11-Rx4) to 0.375 (Sly04-9) with a mean of 0.329. Of the 31 polymorphic SNPs, two SNPs (Sly04-9 and Sly07-2) exhibited the maximum PIC and a gene diversity value of 0.50. The markers Sly11-13 and Sly11-Rx4 were found to be the least informative with PIC and gene diversity values equaling 0.327 and major allele frequency of 0.794. The maximum heterozygosity was observed with SNP markers Sly03-8, Sly06-7, Sly08-1, Sly08-8, while the minimum heterozygosity was observed with SNPs marker Sly02-9, Sly06-1, Sly10-4, Sly10-5, Sly11-13, Sly11-Rx4, and Sly12-9.The SNPs tested for shelf life genes rin (Sly05-rin1 and Sly05-rin2) and nor (Sly10-nor) were monomorphic within the seven different plants (parentals). The SNP (Sly10-alc) tested for the alc gene was however, polymorphic within the seven lines. The result indicates that only one SNP (Sly10-alc) is a functional SNP to detect a long shelf life gene. Tomato genotypes CSIR/CRI-P002 and CSIR/CRI-ATS064 were however, polymorphic for alc. The other SNPs for rin and nor were not useful and new SNPs are needed to be redesigned. The molecular test revealed two superior F1 hybrids which were selected for evaluation as lines for improvement. The advantage is the reduction in cost, time, labor and field space requirement.

Background/Objectives: Understanding the genetic diversity of soybean genotypes can provide valuable information that guides parental selection and the design of an effective hybridization strategy in a soybean breeding program. In order to identify genetically diverse, complementary, and prospective parental lines for breeding, this study set out to ascertain the genetic diversity, relationships, and population structure among 35 soybean genotypes based on agro-morphological traits and Single Nucleotide Polymorphic (SNP) marker data. Methods/Results: Cluster analysis, based on agro-morphological traits, grouped the studied genotypes into four clusters. The first two principal components accounted for 62.8% of the total phenotypic variation, where days to 50% flowering, days to 95% maturity, grain yield, shattering score, and lodging score had high and positive contributions to the total variation. Using the SNP marker information, mean values of 0.16, 0.19, 0.067, and 0.227 were obtained for minor allele frequency (MAF), polymorphic information content (PIC), observed heterozygosity (Ho), and expected heterozygosity (He), respectively. Using different clustering approaches (admixture population structure, principal component scatter plot, and hierarchical clustering), the studied genotypes were grouped into four major clusters. Conclusions:The agro-morphological and molecular analysis results indicated the existence of moderate genetic diversity among the studied genotypes. The traits identified to be significantly related to yield provide valuable information for the genetic improvement of soybeans for yield.

In total, 166 individuals from five indigenous Ethiopian cattle populations – Ambo (n = 27), Borana (n = 35), Arsi (n = 30), Horro (n = 36), and Danakil (n = 38) – were genotyped for 8773 single nucleotide polymorphism (SNP) markers to assess genetic diversity, population structure, and relationships. As a representative of taurine breeds, Hanwoo cattle (n = 40) were also included in the study for reference. Among Ethiopian cattle populations, the proportion of SNPs with minor allele frequencies (MAFs) ≥0.05 ranged from 81.63% in Borana to 85.30% in Ambo, with a mean of 83.96% across all populations. The Hanwoo breed showed the highest proportion of polymorphism, with MAFs ≥0.05, accounting for 95.21% of total SNPs. The mean expected heterozygosity varied from 0.370 in Danakil to 0.410 in Hanwoo. The mean genetic differentiation (FST; 1%) in Ethiopian cattle revealed that within individual variation accounted for approximately 99% of the total genetic variation. As expected, FST and Reynold genetic distance were greatest between Hanwoo and Ethiopian cattle populations, with average values of 17.62 and 18.50, respectively. The first and second principal components explained approximately 78.33% of the total variation and supported the clustering of the populations according to their historical origins. At K = 2 and 3, a considerable source of variation among cattle is the clustering of the populations into Hanwoo (taurine) and Ethiopian cattle populations. The low estimate of genetic differentiation (FST) among Ethiopian cattle populations indicated that differentiation among these populations is low, possibly owing to a common historical origin and high gene flow. Genetic distance, phylogenic tree, principal component analysis, and population structure analyses clearly differentiated the cattle population according to their historical origins, and confirmed that Ethiopian cattle populations are genetically distinct from the Hanwoo breed.

The extent of genetic diversity and relatedness of cowpea germplasm from East Africa are poorly understood. A set of 13 microsatellites (SSR) and 151 single nucleotide polymorphisms (SNPs) markers were applied to assess the levels of genetic diversity in a sample of 95 accessions of local cowpea germplasm and inbred lines of Vigna unguiculata. The average genetic diversity (D), as quantified by the expected heterozygosity, was higher for SSR loci (0.52) than for SNPs (0.34). The polymorphic information content was 0.48 for SSR and 0.28 for SNP while the fixation index was 0.095 for SSR and 0.15 for SNPs showing moderate differentiation and high gene flow among cowpea accessions from East African countries. The results of data analysis of both SSR and SNP markers showed similar clustering patterns suggesting a substantial degree of association between origin and genotype. Principal coordinate analysis (PCoA) with SSR and SNP markers showed that accessions were grouped into two and three broad groups across the first two axes, respectively. Our study found that SNP markers were more effective than SSR in determining the genetic relationship among East African local cowpea accessions and IITA inbred lines. Based on this analysis, five local cowpea accessions Tvu-13490, Tvu-6378, Tvu-13448, Tvu-16073, and 2305675 were identified to be tightly clustered sharing several common alleles with the drought tolerant variety Danila when analyzed with SSR and SNP markers. The findings will assist and contribute to future genetic diversity studies aimed at the genetic improvement of local Eastern Africa cowpea accessions for improved overall agronomic performance in general and breeding for drought tolerant in particular.