Informative Simple Sequence Repeat Research Articles

Genetic diversity analysis of Sinojackia microcarpa, a rare tree species endemic in China, based on simple sequence repeat markers

Although most Sinojackia species are endangered, they contribute greatly to the biodiversity of local ecosystems. Sinojackia microcarpa, an endangered species, is distributed only in three provinces of eastern China. Determining the genetic diversity of S. microcarpa provides key information for germplasm evaluation and species conservation. Here we used simple sequence repeat (SSR) markers to investigate the genetic diversity of eight natural populations of S. microcarpa. Leaf samples were collected from 144 individuals in 8 wild populations. The 156 bands were generated from 14 pairs of informative SSR primers, with an average percentage of polymorphic bands of 45.67%. The average values of Nei’s genetic diversity (He) and Shannon’s diversity index (I) were 0.1007 and 0.1658, respectively. The total genetic variation of S. microcarpa existed mainly within the eight populations, rather than among populations, and reached 86.41%. A cluster analysis showed that the eight wild populations of S. microcarpa could be classified into four groups, at a threshold of 4.0, based on an analysis of the SSR genotypes. Furthermore, there was a significant association between the phylogenetic relationships and the geographic locations of the S. microcarpa populations. In particular, populations from Fuyang, Jiande, and Lin’an in Zhejiang Province had close phylogenetic relationships and geographic distances. In addition, these three populations had the highest genetic diversity and the most individuals, suggesting that these three locations may be the S. microcarpa distribution center. This study serves as a model for studying the genetic diversity of endangered plant species.

Genetic Variation and Association Analysis of the SSR Markers Linked to the Major Drought-Yield QTLs of Rice.

Drought is one of the major abiotic stresses, which hampers the production of rice worldwide. Informative molecular markers are valuable tools for improving the drought tolerance in various varieties of rice. The present study was conducted to evaluate the informative simple sequence repeat (SSR) markers in a diverse set of rice genotypes. The genetic diversity analyses of the 83 studied rice genotypes were performed using 34 SSR markers closely linked to the major quantitative trait loci (QTLs) of grain yield under drought stress (qDTYs). In general, our results indicated high levels of polymorphism. In addition, we screened these rice genotypes at the reproductive stage under both drought stress and nonstressful conditions. The results of the regression analysis demonstrated a significant relationship between 11 SSR marker alleles and the plant paddy weight under stressful conditions. Under the nonstressful conditions, 16 SSR marker alleles showed a significant correlation with the plant paddy weight. Finally, four markers (RM279, RM231, RM166, and RM231) demonstrated a significant association with the plant paddy weight under both stressful and nonstressful conditions. These informative-associated alleles may be useful for improving the crop yield under both drought stress and nonstressful conditions in breeding programs.

Development of User-Friendly Method to Distinguish Subspecies of the Korean Medicinal Herb Perilla frutescens Using Multiplex-PCR.

Perilla (Perilla frutescens) is an economically and culturally important plant in East Asia. Plant breeding between cultivars has enhanced the genetic diversity of perilla overall, but means that functionally diverse subspecies are more difficult to identify and distinguish. In this study, we developed gene-based DNA markers to distinguish between the Korean herbal medicinal perilla varieties. We identified informative simple sequence repeat (SSR) regions on the promoter regions of the Myb-P1 and dihydroflavonol 4-reductase (DFR) genes, as well as a large insertion-deletion (indel) region in the limonene synthase (LS) gene, and developed markers to characterize the distinct subspecies differences (PfMyb-P1pro, PfDFRpro, and PfLS, respectively). Using the PfLS primers, a 430-bp region could be amplified from P. frutescens var. acuta, crispa, and f. viridis (known as Jasoyeop, Jureum-soyeop, and Chungsoyeop, respectively), but not from P. frutescens var. japonica (Dlggae). The PfMybpro primers resulted in PCR products of 314 or 316, 330, 322, and 315 bp from Dlggae, Jasoyeop, Jureum-soyeop, and Chungsoyeop, respectively, and the PfDFRpro primers resulted in products of 189 or 202, 187 or 189, 185 or 189, and 193bp, respectively, for the four perilla subspecies. Combining these three reactions into a single multiplex PCR approach resulted in subspecies-specific PCR band patterns for six common types of commercial perilla, distinguishing between three varieties of Dlggae (Cham-Dlggae, Ip-Dlggae, and Bora-Dlggae), as well as identifying Jasoyeop, Jureum-soyeop, and Chungsoyeop. These user-friendly markers will be valuable as a simple and efficient method for identifying the Korean medicinal herb Jasoyeop, as well as distinguishing between other functionally distinct subspecies, which may have broad applications in the Korean herbal industry.

Genetic diversity assessment of a winter squash and pumpkin (Cucurbita maxima Duchesne) germplasm collection based on genomic Cucurbita-conserved SSR markers

Molecular marker examinations of genetic diversity are valuable for assessment of within-species diversity and relationships, genotype identification, to assist selection for crop improvement programs, and to handle coherently germplasm collections. Cucurbita maxima is a highly diverse cucurbit crop, but its genetic diversity at the molecular level is inadequately characterized. Presently, genomic simple sequence repeat (SSR) markers that were described and employed for Cucurbita moschata and Cucurbita pepo were screened on C. maxima. A set of 23 highly informative SSRs was selected to evaluate genetic diversity within a collection of 85 C. maxima accessions. This collection contained cultivars and breeding lines from Europe, North America, Asia, Australia, and New Zealand, of six horticultural groups. Based on the genotyping results and statistical analyses, a fundamental division was observed in C. maxima above the level of horticultural groups. One major cluster was comprised between accessions from the Banana, Buttercup, and Hubbard horticultural groups. The other major cluster was divided in two sub-clusters, the larger of which included all accessions of the Mammoth (Show) Pumpkin horticultural group and the smaller, more outlying cluster was comprised of accessions of the Australian Blue horticultural group, with the one representative of the Turban group most outlying. Generally, the horticultural groups formed sub-sub-clusters, as most accessions within these respective groups showed close affinity. The selected set of 23 Cucurbita-conserved SSR markers is expected to be of great value for further germplasm characterization in this species and for taxonomical identifications within the genus Cucurbita.

Cross-taxon application of sugarcane EST-SSR to genetic diversity analysis of bermudagrass (Cynodon spp.)

Bermudagrass lags in genomic and molecular breeding resources, particularly regarding a critical mass of robust, reproducible, and highly polymorphic molecular markers like simple sequence repeats (SSR). Here, 2017 sugarcane EST-SSR primer pairs (PPs) were screened for transferability and polymorphisms against commercial triploid hybrids and representatives of their parental species, tetraploid Cynodon dactylon and diploid C. transvaalensis. Fifty-four percent of PPs amplified target SSR in at least one of the two species, while 62% of these ‘transferable’ SSR were polymorphic. A subset of 228 polymorphic markers was utilized for genotyping 24 samples that included: 10 ‘Tif’ series cultivars (i.e., released from Tifton, GA), their parental species, 10 F1 hybrids between the two species, and two diverse Sorghum spp. A total of 90 and 144 PPs produced reproducible bands in the bermudagrass samples and the sorghum species, respectively. While 63 (70%) PPs were polymorphic among members of the ‘Tif’ diversity panel, 79 (54.8%) were polymorphic between the sorghum species. Further, 57 PPs polymorphic in the ‘Tif’ diversity panel were genotyped against 3 experimental lines and 7 commercial hybrids including ‘TifTuf’, a recent addition to the ‘Tif’ series. Joint analysis of 24 genotypes grouped bermudagrass accessions into two main clusters, one including C. dactylon and the other including C. transvaalensis. A small set of informative SSR readily differentiated the cultivar groups and identified potentially mischaracterized cultivars, but did not differentiate among mutants within cultivar groups. Heterologous EST-SSR from sugarcane are useful sources of polymorphic markers for cultivar identification in bermudagrass, and build on existing marker resources to facilitate genetic mapping, QTL analysis, and marker assisted selection.

Development of genome-wide SSR markers in melon with their cross-species transferability analysis and utilization in genetic diversity study

Simple sequence repeats (SSRs) are one of the most informative and widely used molecular markers in plant research. The melon draft genome has provided a powerful tool for SSR marker development in this species in which there are still not enough SSR markers. We therefore developed genome-wide SSR markers from melon, which were used for genetic diversity analysis in melon accessions and comparative mapping with cucumber and watermelon. A total of 44,265 microsatellites from the melon genome were characterized, of which 28,570 SSR markers were developed. In silico PCR analysis with these SSR markers identified 4002 and 1085 with one amplicon in cucumber and watermelon genome, respectively. With these cross-species transferable melon SSR markers, the chromosome synteny between melon and cucumber as well as watermelon was established, which revealed complicated mosaic patterns of syntenic blocks among them. We experimentally validated 384 SSR markers, from which 42 highly informative SSR ones were selected for genetic diversity and population structure analysis among 118 melon accessions. The large number of melon SSR markers developed in this study provides a valuable resource for genetic linkage map construction, molecular mapping, and marker-assisted selection (MAS) in melon. Furthermore, the cross-species transferable SSR markers could also be useful in various molecular marker-related studies in other closely related species in Cucurbitaceae family in which draft genomes are not yet available.

An Efficient Strategy Combining SSR Markers- and Advanced QTL-seq-driven QTL Mapping Unravels Candidate Genes Regulating Grain Weight in Rice.

Development and use of genome-wide informative simple sequence repeat (SSR) markers and novel integrated genomic strategies are vital to drive genomics-assisted breeding applications and for efficient dissection of quantitative trait loci (QTLs) underlying complex traits in rice. The present study developed 6244 genome-wide informative SSR markers exhibiting in silico fragment length polymorphism based on repeat-unit variations among genomic sequences of 11 indica, japonica, aus, and wild rice accessions. These markers were mapped on diverse coding and non-coding sequence components of known cloned/candidate genes annotated from 12 chromosomes and revealed a much higher amplification (97%) and polymorphic potential (88%) along with wider genetic/functional diversity level (16–74% with a mean 53%) especially among accessions belonging to indica cultivar group, suggesting their utility in large-scale genomics-assisted breeding applications in rice. A high-density 3791 SSR markers-anchored genetic linkage map (IR 64 × Sonasal) spanning 2060 cM total map-length with an average inter-marker distance of 0.54 cM was generated. This reference genetic map identified six major genomic regions harboring robust QTLs (31% combined phenotypic variation explained with a 5.7–8.7 LOD) governing grain weight on six rice chromosomes. One strong grain weight major QTL region (OsqGW5.1) was narrowed-down by integrating traditional QTL mapping with high-resolution QTL region-specific integrated SSR and single nucleotide polymorphism markers-based QTL-seq analysis and differential expression profiling. This led us to delineate two natural allelic variants in two known cis-regulatory elements (RAV1AAT and CARGCW8GAT) of glycosyl hydrolase and serine carboxypeptidase genes exhibiting pronounced seed-specific differential regulation in low (Sonasal) and high (IR 64) grain weight mapping parental accessions. Our genome-wide SSR marker resource (polymorphic within/between diverse cultivar groups) and integrated genomic strategy can efficiently scan functionally relevant potential molecular tags (markers, candidate genes and alleles) regulating complex agronomic traits (grain weight) and expedite marker-assisted genetic enhancement in rice.

Novel and highly informative Capsicum SSR markers and their cross-species transferability.

This study was undertaken primarily to develop new simple sequence repeat (SSR) markers for Capsicum. As part of this project aimed at broadening the use of molecular tools in Capsicum breeding, two genomic libraries enriched for AG/TC repeat sequences were constructed for Capsicum annuum. A total of 475 DNA clones were sequenced from both libraries and 144 SSR markers were tested on cultivated and wild species of Capsicum. Forty-five SSR markers were randomly selected to genotype a panel of 48 accessions of the Capsicum germplasm bank. The number of alleles per locus ranged from 2 to 11, with an average of 6 alleles. The polymorphism information content was on average 0.60, ranging from 0.20 to 0.83. The cross-species transferability to seven cultivated and wild Capsicum species was tested with a set of 91 SSR markers. We found that a high proportion of the loci produced amplicons in all species tested. C. frutescens had the highest number of transferable markers, whereas the wild species had the lowest. Our results indicate that the new markers can be readily used in genetic analyses of Capsicum.

Analysis of genetic diversity and population structure of peanut cultivars and breeding lines from China, India and the US using simple sequence repeat markers.

Cultivated peanut is grown worldwide as rich-source of oil and protein. A broad genetic base is needed for cultivar improvement. The objectives of this study were to develop highly informative simple sequence repeat (SSR) markers and to assess the genetic diversity and population structure of peanut cultivars and breeding lines from different breeding programs in China, India and the US. A total of 111 SSR markers were selected for this study, resulting in a total of 472 alleles. The mean values of gene diversity and polymorphic information content (PIC) were 0.480 and 0.429, respectively. Country-wise analysis revealed that alleles per locus in three countries were similar. The mean gene diversity in the US, China and India was 0.363, 0.489 and 0.47 with an average PIC of 0.323, 0.43 and 0.412, respectively. Genetic analysis using the STRUCTURE divided these peanut lines into two populations (P1, P2), which was consistent with the dendrogram based on genetic distance (G1, G2) and the clustering of principal component analysis. The groupings were related to peanut market types and the geographic origin with a few admixtures. The results could be used by breeding programs to assess the genetic diversity of breeding materials to broaden the genetic base and for molecular genetics studies.

Development of genome-wide informative simple sequence repeat markers for large-scale genotyping applications in chickpea and development of web resource.

Development of informative polymorphic simple sequence repeat (SSR) markers at a genome-wide scale is essential for efficient large-scale genotyping applications. We identified genome-wide 1835 SSRs showing polymorphism between desi and kabuli chickpea. A total of 1470 polymorphic SSR markers from diverse coding and non-coding regions of the chickpea genome were developed. These physically mapped SSR markers exhibited robust amplification efficiency (73.9%) and high intra- and inter-specific polymorphic potential (63.5%), thereby suggesting their immense use in various genomics-assisted breeding applications. The SSR markers particularly derived from intergenic and intronic sequences revealed high polymorphic potential. Using the mapped SSR markers, a wider functional molecular diversity (16–94%, mean: 68%), and parentage- and cultivar-specific admixed domestication pattern and phylogenetic relationships in a structured population of desi and kabuli chickpea genotypes was evident. The intra-specific polymorphism (47.6%) and functional molecular diversity (65%) potential of polymorphic SSR markers developed in our study is much higher than that of previous documentations. Finally, we have developed a user-friendly web resource, Chickpea Microsatellite Database (CMsDB; http://www.nipgr.res.in/CMsDB.html), which provides public access to the data and results reported in this study. The developed informative SSR markers can serve as a resource for various genotyping applications, including genetic enhancement studies in chickpea.

Genomic composition and yield heterosis of the partial inter-specific hybrid rice between Oryza sativa L. and Oryza glaberrima Steud.

SUMMARYDevelopment of partial inter-specific hybrid (PIH) rice is a promising approach for exploiting inter-specific heterosis between Oryza glaberrima and Oryza sativa. In the present study, the relationship between genetic distance (GD) of parental lines and yield performance of the PIHs was assessed using partial diallel crosses between three indica male sterile lines and 14 introgression lines (ILs) with different fragments of O. glaberrima genes. Twenty two out of the 42 PIHs expressed positive heterobeltiosis (i.e., the Fl hybrid showed superiority over the better parent in the target trait) for panicle number, spikelet number, thousand grain weight and grain yield/plant. The proportion of O. glaberrima genome in the ILs ranged from 0·03 to 0·41, as revealed by 16 informative simple sequence repeat markers. Significant positive correlations were found between the proportion of O. glaberrima genome of the ILs and the GD between the ILs and the three different female parents. Heterosis of spikelet number per panicle in the hybrids was positively proportional with the O. glaberrima genome content of the parental ILs, while that of fertile grain percentage was negatively proportional to the O. glaberrima genome proportion. On average, the PIHs with higher grain yield and highest heterobeltiosis were obtained from the ILs carrying between 0·15 and 0·30 of O. glaberrima genome. The results indicated that a small proportion (<0·15) of the O. glaberrima genome in the ILs might limit heterosis expression of spikelet number per panicle, while a very large proportion (>0·30) of the O. glaberrima genome decreased the grain filling percentage in the PIHs, thus an intermediate range of O. glaberrima genome proportion should be more suitable for breeding heterotic PIHs. The exploitation of inter-specific heterosis between O. glaberrima and O. sativa has potential value for heterotic breeding in rice.

Microsatellite resources of Eucalyptus: current status and future perspectives.

Eucalyptus is the premier paper pulp, short rotation plantation species grown all over the world. Genetic improvement programs integrating molecular marker tools are in progress in many parts of the globe to increase the productivity. Whole genome sequence and expressed sequence tags (ESTs) of the eucalypts paved way for introduction of molecular genetics and breeding in this genus. Different molecular characterization approaches have been used simultaneously in eucalypts, however, microsatellites or simple sequence repeats (SSRs) with their prolific characteristics could occupy a special niche in Eucalyptus genetic improvement. Further, highly informative SSRs were used for the clonal identity, genetic fidelity and in certification of breeder’s rights. Eucalyptus genetic linkage maps generated with microsatellite loci were used successfully to identify quantitative trait loci (QTLs) for various economically important traits. Progressively more numbers of microsatellites are being linked to genes associated with adaptive and functional variations, therefore making their utility broader in genetic applications. Availability of common SSR markers across the species provides an opportunity to validate the expression of QTLs across variable genetic backgrounds and accurately compare the position of QTLs in other species. Recent evidences suggest that the presence of SSRs in micro RNAs of plant species play a role in the quantitative trait expression. Similar studies in eucalypts may provide new insights into the genetic architecture of transcript-level variations and post transcriptional gene regulation. This review on eucalypts microsatellites, highlights the availability and characteristics of genomic and eSSRs and their potential in genetic analysis of natural and breeding populations and also discusses the future prospects in population genetics and marker assisted selection.Electronic supplementary materialThe online version of this article (doi:10.1186/s40529-014-0073-3) contains supplementary material, which is available to authorized users.

Thai elite cassava genetic diversity was fortuitously conserved through farming with different sets of varieties

Many in situ conservation programs have been developed to preserve plant landrace diversity and to promote its sustainable utilization, but little is known about the effectiveness of the developed programs in conserving plant genetic diversity. We investigated the effectiveness of an unregulated (i.e., unplanned or open) conservation system maintained by Thai farmers in conserving Thai elite cassava (Manihot esculenta Crantz) varieties. Specifically, we compared genetic diversity of 266 cassava clones that were collected from 80 farms in eight provinces with 16 cassava landraces and varieties released since the 1970s through genotyping with 35 informative simple sequence repeat (SSR) markers. The SSR analysis revealed a large regional heterogeneity in cassava diversity, with a strong genetic differentiation of the assayed clones among the 80 farms (19.8 %) and across the eight provinces (11.8 %). Significant associations were also found between SSR variation and farm agro-ecological factors or some farming practices. However, there was no significant genetic differentiation (0.9 %) between the 266 farm clones and 16 reference varieties. These findings suggest that the Thai elite cassava genetic diversity was fortuitously conserved by the farmers through farming with different sets of varieties. Implications of these findings are discussed with respect to on-farm conservation of plant genetic resources.

Plant morphology, genome size, and SSR markers differentiate five distinct taxonomic groups among accessions in the genus Miscanthus

AbstractInformation on genome size, ploidy level, and genomic polymorphisms among accessions of the genus Miscanthus can assist in taxonomic studies, help understand the evolution of the genus, and provide valuable information to biomass crop improvement programs. Taxonomic investigation combining variation in plant morphology, genome size, chromosome numbers, and simple sequence repeat (SSR) marker polymorphisms were applied to characterize 101 Miscanthus accessions. A total of 258 amplicons generated from 17 informative SSR primer pairs was subjected to cluster and principal coordinate analysis and used to characterize genetic variation and relationships among 31 Miscanthus accessions, including four interspecific Miscanthus hybrids created from controlled pollinations, and four Saccharum, six Erianthus, and one Sorghum bicolor accessions. Miscanthus accessions were distinct from accessions in the genera Erianthus and Saccharum. Miscanthus accessions fell into five taxonomic groups, including the existing taxonomic section Miscanthus, diploid and tetraploid Miscanthus sacchariflorus, and a fourth (M. × giganteus) and fifth group (Miscanthus ‘purpurascens’); the last two being intermediate forms. In contrast to previous work, our findings suggest diploid and tetraploid M. sacchariflorus are taxonomically different, the latter more closely related to M. sacchariflorus var lutarioriparius. We also suggest that Miscanthus ‘purpurascens’ accessions are interspecific hybrids between Miscanthus sinensis and diploid M. sacchariflorus based on DNA content and SSR polymorphisms. The evolution of Miscanthus and related genera is discussed based on combined analysis and geographical origin.

Hybrid purity assessment in Eucalyptus F1 hybrids using microsatellite markers.

The worldwide expansion of hybrid breeding and clonal forestry is to meet the demands of paper pulp and bioenergy. Although India was one of the pioneers in hybrid production of eucalypts only recently the hybrid clonal forestry is gaining momentum. Inter-specific hybrids are being produced to exploit the hybrid vigor of F1 individuals. Quality control genotyping for hybrid purity and parentage confirmation at the early stage is one of the essential criteria for clonal propagation and field trails for the assessment of growth performance. Eucalyptus being a obligatory outcrossed species with potential to self pollination, possibilities of pollen contamination are high. Hence, in the present study, Eucalyptus camaldulensis × E. tereticornis inter-specific hybrids were genotyped using 25 fluorescent labeled microsatellite markers available in public domain. Multiplex loading of PCR products was performed successfully for most of the microsatellite loci. Hybrid purity index was calculated and parentage was confirmed. Hybrid purity values ranged from 85 to 100 % showed the efficiency of controlled pollination techniques. A subset of six fully informative simple sequence repeats was identified for routine quality control genotyping for these hybrids. Detection of non-essential genotypes observed among the hybrid seedlings proved the significance of hybrid purity tests and the false hybrids were removed at the seedling stage. The hybrids with proven hybridity will be used for generation of genetic linkage, discovery of quantitative trait loci and the individuals with high productivity can enter into mass clonal multiplication.

Genetic Structure in a Core Subset of Cultivated Barley Germplasm

ABSTRACTCultivated barley (Hordeum vulgare L. subsp. vulgare) is one of the economically most important cereals with a large number of germplasm accessions conserved worldwide. A core subset of 571barley accessions representing 94 countries was developed, genotyped using 30 informative simple sequence repeat (SSR) markers, and analyzed with several common approaches. The distance‐based analysis of the 230 SSR allele profile revealed a major division of the barley accessions that explained 6 to 8% SSR variation. The model‐based Bayesian analyses revealed six optimal clusters that captured 17.5% SSR variation. The clusters varied in size from 34 to 160, consisted of accessions from many countries and regions, and were well represented by accessions with various important agronomic traits such as kernel row number and growth habit. Such germplasm partitions were further supported from principal component and neighbor‐joining analyses. A mini core subset of 219 accessions from 94 countries was thus developed for future structural inferences. Interestingly, the revealed genetic structure is consistent with the known barley domestication history. Therefore, the inferred optimal clusters may provide the best possible structural description of the current global cultivated barley gene pool and also are useful for future barley research in association mapping, adaptation, and domestication.

Identification of informative simple sequence repeat (SSR) markers for drought tolerance in maize

Maize is moderately sensitive to drought. Drought affects virtually all aspects of maize growth in varying degrees at all stages, from germination to maturity. Tolerance to drought is genetically and physiologically complicated and inherited quantitatively. Application of molecular-marker aided selection technique for improvement of drought tolerance would accelerate breeding progress by increasing selection efficiency. One of the most important aims in plant breeding is to determine the chromosomal regions related to drought tolerance. Therefore, 38 maize hybrids were planted in two separate experiments with well-watered (WW) and water-stressed (WS) conditions at grain filling period using a randomized complete block design (RCBD) with three replications. Drought tolerance indices such as stress susceptibility index (SSI), mean productivity (MP), tolerance (TOL) and stress tolerance index (STI) were used to evaluate the susceptibility and tolerance of the hybrids. Also, to assess the genetic relationships among the 38 maize hybrids used in the evaluation and breeding for drought tolerance, and to determine informative markers for drought tolerance, 12 microsatellite primers were used. Genomic DNA was extracted with the CTAB method and PCR was performed based on the common method for microsatellite markers. PCR products were separated using 6% polyacrylamide denaturing gel. Stepwise multiple regression analysis was used to determine the chromosomal regions related to drought tolerance. A total of 40 simple sequence repeat (SSR) alleles (bands) with a mean of 3.33 alleles per locus were identified. Polymorphism information content (PIC) of the 12 SSR loci ranged from 0.23 (Phi080) to 0.79 (UMC2359), with a mean PIC of 0.53. The analysis also led to identification of informative SSR markers, namely UMC1862 (bin 1.11), UMC1719 (bin 4.10-4.11), UMC1447 (bin 5.03), UMC2359 (bin 9.07) and UMC1432 (bin 10.02), which significantly contributed to the differentiation of the drought tolerant and susceptible genotypes analyzed in the study. These SSR markers could be further validated and potentially deployed in molecular marker-assisted breeding for drought tolerance in maize.

Identification and validation of a core set of microsatellite markers for genetic diversity analysis in watermelon, Citrullus lanatus Thunb. Matsum. & Nakai

Watermelon, Citrullus lanatus Thunb. Matsum. & Nakai is an important vegetable crop worldwide. Due to its narrow genetic base, detection and utilization of the genetic variations, cultivar identification and increasing genetic diversity are some important tasks for watermelon breeders. Molecular markers, especially microsatellites or simple sequence repeats (SSRs) are playing increasingly important roles for these purposes. In the present study, a core set of 23 highly informative SSR markers was developed for watermelon genetic diversity analysis. Based on whole genome sequencing of 17 watermelon inbred lines, we identified 3.9 million single nucleotide polymorphisms (SNPs) which were used to construct a SNP-based dendrogram for the 17 lines. Meanwhile, from the sequenced genome, 13,744 SSRs were developed, of which 704 were placed on a high-resolution watermelon linkage map. To develop the core set SSR markers, 78 of the 704 mapped SSRs were selected as the candidate markers. Using the SNP-based dendrogram as calibration, 23 SSR markers evenly distributed across the genome were identified as the core marker set for watermelon genetic diversity analysis. Each marker was able to detect 2–7 alleles with polymorphism information content values ranging from 0.45 to 0.82. The dendrograms of 17 watermelon lines based on SNPs, the base set of 78 SSRs and the core set of 23 SSRs were highly consistent. The utility of this core set SSRs was demonstrated in 100 commercial watermelon cultivars and elite lines, which could be placed into six clusters that were largely consistent with previous classification based on morphology and parentage data. This core set of SSR markers should be very useful for genotyping and genetic variation analysis in watermelon.

Biotechnology for abiotic tolerance and nutritional improvement in selected genotypes of Solanum melongena and Cucurbita moschata

Molecular marker examinations of genetic diversity are valuable for assessment of within-species diversity and relationships, genotype identification, to assist selection for crop improvement programs, and to handle coherently germplasm collections. Cucurbita maxima is a highly diverse cucurbit crop, but its genetic diversity at the molecular level is inadequately characterized. Presently, genomic simple sequence repeat (SSR) markers that were described and employed for Cucurbita moschata and Cucurbita pepo were screened on C. maxima. A set of 23 highly informative SSRs was selected to evaluate genetic diversity within a collection of 85 C. maxima accessions. This collection contained cultivars and breeding lines from Europe, North America, Asia, Australia, and New Zealand, of six horticultural groups. Based on the genotyping results and statistical analyses, a fundamental division was observed in C. maxima above the level of horticultural groups. One major cluster was comprised between accessions from the Banana, Buttercup, and Hubbard horticultural groups. The other major cluster was divided in two sub-clusters, the larger of which included all accessions of the Mammoth (Show) Pumpkin horticultural group and the smaller, more outlying cluster was comprised of accessions of the Australian Blue horticultural group, with the one representative of the Turban group most outlying. Generally, the horticultural groups formed sub-sub-clusters, as most accessions within these respective groups showed close affinity. The selected set of 23 Cucurbita-conserved SSR markers is expected to be of great value for further germplasm characterization in this species and for taxonomical identifications within the genus Cucurbita.

Identification of sugarcane microsatellites associated to sugar content in sugarcane and transferability to other cereal genomes

Microsatellites or simple sequence repeats (SSRs) markers are very informative for various applications in genetics and breeding. Information obtained with these markers has contributed to a better understanding of evolution and the complexity of the sugarcane genome. With the objective of identifying a large set of polymorphic microsatellite markers designated as Unigene derived Sugarcane Microsatellite (UGSM) and Sugarcane Enriched Genomic Microsatellite (SEGMS), 351 UGSM and 36 SEGMS were tested to find out informative SSRs marker for sugar content. These markers were screened and validated for their use in genetic diversity, cross transferability and comparative linkage potential in high and low sugar bulk of two segregating progenies and twenty each, cultivated high and low sugar cultivars. 158 (40.83%) of the microsatellite markers (144-UGSM: 14-SEGMS) were found to be highly robust and polymorphic. Cross amplification was estimated among nineteen accessions of six sugarcane cultivars, one inter specific hybrids, five related species, four related genera, and three divergent genera by using 27 UGSM primers. Analysis of 388 alleles, amplified by these markers, indicated the high number of observed allele ranged from 2 to 26, with an average of 14.37 alleles detected per locus. High level of polymorphism detected by these markers among sugarcane species, genera and cultivars was 96.3%, while cross-transferability rate was 98.0% within Saccharum complex and 88.27% to cereals. Wide range of genetic diversity (0.33–0.79 with an average of 0.56) assayed with UGSM markers suggested their importance in various genotypic applications in sugarcane.