Simple Sequence Repeat Markers Research Articles

SSR marker-based genetic diversity and marker-trait association analysis in aromatic rice (Oryza sativa) landraces

The present study on SSR marker-based genetic diversity and marker-trait association analysis in aromatic rice (Oryza sativa L.) landraces was carried out at R. H. Richharia Research Laboratory, Indira Gandhi Krishi Vishwavidyalaya, Raipur, Chhattisgarh during 2020. A total of 25 PCR-based simple sequence repeats (SSR) markers were evaluated in a set of 90 aromatic rice landraces along with 6 checks which includes 1 non-aromatic and 5 aromatic check varieties. Phenotypic data for marker-trait association analysis were taken for 24 yield attributing traits. Polymorphic Information Content (PIC) value ranged from 0.52 (RM316) to 0.79 (RM553) with a mean of0.69 which reveals that all the markers used in this study were highly informative and useful for diversity analysis of a wide range of genotypes. Unweighted Pair Group Method with Arithmetic Mean (UPGMA) and Rho’s similarity based cluster analysis revealed that all 5 aromatic check varieties falls in one cluster while the 1 non-aromatic check variety (Mahamaya) forms a separate cluster. Mixed linear model (MLM) was applied to perform genome-wide association mapping where 41 significant marker-trait associations were observed for 24 yield attributing traits. The potential markers identified in the study may provide new opportunities for rice breeder to improve yield and its attributing traits through marker assisted selection approach.

Characterization of the genetic diversity of Citrus species of Nepal using simple sequence repeat (SSR) markers

Characterization of the genetic diversity of Citrus species of Nepal using simple sequence repeat (SSR) markers

Microsatellite marker assisted molecular diversity and population structure analysis of Kaempferia galanga Linn. germplasm collected from different parts of India

Kaempferia galanga Linn. is an endangered medicinal and aromatic crop with a wide range of importance in the field of pharmaceutical industries. The current study is sought to examine the molecular variance and population diversity for the first time within the core collection of K. galanga germplasm. For this study, a total of 132 accessions of K. galanga were collected from various states of India and all the germplasm has been maintained ex-situ at the experimental farm of CSIR-NEIST, Jorhat. Initially, 58 simple sequence repeats (SSR) markers were tested among them 48 markers that were the most efficient, sensitive, with positive predictive and polymorphism were used in the study. The population structure analysis of K. galanga showed nine clusters based on the Bayesian model. The estimated population heterozygosity ranged from 0.117 to 0.250. According to the Analysis of molecular variance study (AMOVA), the variation within the population (68%) was more than among the population (32%). The genetic diversity among the population (Fst) was in the range of 0.28–0.60. The accessions under study demonstrated a high amount of polymorphism with a 99.4% polymorphism observed in molecular investigation. The outcome of the study would help in the identification of the genetic variance for unique germplasm which would be of great aid in the breeding program of K. galanga.

황해쑥 품종 구별을 위한 유전체 염기서열 기반 genomic SSR marker 개발

Background: Artemisia argyi is an economically important medicinal plant with in the genus of Artemisia. However, the lack of the molecular markers hinders the understanding of genetic diversity and cultivar discrimination. Therefore, the main objectives of this study were to characterize the SSRs of Artemisia argyi in the reference genome, and to developed polymorphic SSRs for cultivar discrimination and population genetic studies.BRMethods and Results: Novel 100 simple sequence repeat (SSR) markers were designed based on the whole-genome sequence data of Artemisia argyi, and selected 78 polymorphic markers were selected. Upon the assessment of the five accessions with the full set of 78 SSR, the major allele frequency (MAF) ranged from 0.20 to 1, with an average of 0.36. In addition, the number of alleles per locus ranged from 2 to 9 alleles with a mean value of 5.2 alleles per locus. The polymorphism information content (PIC) values ranged from 0.27 to 0.87 with a mean PIC value of 0.69.BRConclusions: The Artemisia argyi alleles were detected in 5 accessions, among them six primers (argyi-gSSR003, argyi-gSSR014, argyi-gSSR035, argyi-gSSR036, argyi-gSSR093, argyigSSR094) produced five cultivar-specific loci, therefore very promising for cultivar identification, genetic diversity and breeding studies in Artemisia argyi.

Genetic diversity of colonial bentgrass Agrostis capillaris based on simple sequence repeat markers and high‐resolution melt analysis with haplotype scoring

AbstractColonial bentgrass (Agrostis capillaris) is a tetraploid rhizomatous bentgrass that generally exhibits improved resistance to the important fungal disease dollar spot and has more tolerance to water‐deficit stress than the more widely grown creeping bentgrass (Agrostis stolonifera). Interspecific hybridization between these two species is possible and efforts to understand the genetic mechanisms of enhanced biotic and abiotic stress resistance in colonial bentgrass are proceeding. To gain a better understanding of the level of genetic diversity in cultivated colonial bentgrasses and to determine if genotypes with improved interspecific hybridization potential can be identified, nine colonial bentgrass cultivars and one creeping bentgrass cultivar were screened with 48 colonial bentgrass‐derived simple sequence repeat markers. Twenty‐one primer pairs were found to produce reliable amplification, and these markers were scored using high‐resolution melt analysis to generate haplotypes that were used to evaluate cultivar relationships. The nine colonial bentgrass cultivars could be placed into two groups and overall showed limited levels of diversity. The creeping bentgrass cultivar was distinct and individual genotypes within each species exhibited haplotypes more common in the alternate species, suggesting that these markers may be useful for selecting genotypes with enhanced interspecific hybridization potential.

Phenotypic effects of Am genomes in nascent synthetic hexaploids derived from interspecific crosses between durum and wild einkorn wheat.

Allopolyploid speciation is a major evolutionary process in wheat (Triticum spp.) and the related Aegilops species. The generation of synthetic polyploids by interspecific crosses artificially reproduces the allopolyploidization of wheat and its relatives. These synthetic polyploids allow breeders to introduce agriculturally important traits into durum and common wheat cultivars. This study aimed to evaluate the genetic and phenotypic diversity in wild einkorn Triticum monococcum ssp. aegilopoides (Link) Thell., to generate a set of synthetic hexaploid lines containing the various Am genomes from wild einkorn, and to reveal their trait characteristics. We examined the genetic diversity of 43 wild einkorn accessions using simple sequence repeat markers covering all the chromosomes and revealed two genetically divergent lineages, L1 and L2. The genetic divergence between these lineages was linked to their phenotypic divergence and their habitats. L1 accessions were characterized by early flowering, fewer spikelets, and large spikelets compared to L2 accessions. These trait differences could have resulted from adaptation to their different habitats. We then developed 42 synthetic hexaploids containing the AABBAmAm genome through interspecific crosses between T. turgidum cv. Langdon (AABB genome) as the female parent and the wild einkorn accessions (AmAm genome) as the male parents. Two of the 42 AABBAmAm synthetic hexaploids exhibited hybrid dwarfness. The phenotypic divergence between L1 and L2 accessions of wild einkorn, especially for days to flowering and spikelet-related traits, significantly reflected phenotypic differences in the synthetic hexaploids. The differences in plant height and internodes between the lineages were more distinct in the hexaploid backgrounds. Furthermore, the AABBAmAm synthetic hexaploids had longer spikelets and grains, long awns, high plant heights, soft grains, and late flowering, which are distinct from other synthetic hexaploid wheat lines such as AABBDD. Utilization of various Am genomes of wild einkorn resulted in wide phenotypic diversity in the AABBAmAm synthetic hexaploids and provides promising new breeding materials for wheat.

Assessing the Genetic Diversity of Daylily Germplasm Using SSR Markers: Implications for Daylily Breeding.

This work aims to characterize the genetic diversity of species, early hybrids, and cultivars using microsatellite simple sequence repeat (SSR) markers, as well as analyze and identify the origin of Hemerocallis spp. early hybrids. For this research, samples were collected from different types of daylily species, early hybrids (known or hypothetically first-generation hybrids from Hemerocallis species), foreign, and Lithuanian varieties. An initial screening of SSR primers developed for Hemerocallis citrina was performed, and their suitability for testing other daylily species and hybrids was evaluated. The genetic diversity was assessed with the selected eight-primer set, and molecular SSR profiles were created. Primer SAU00097 is the most informative according to heterozygosity (0.95) and polymorphism information content (PIC) (0.17). The highest heterozygosity was observed in Lithuanian cultivars (0.713), the lowest in species (0.583). Genetic relationships between species show that only fulvous daylilies are separated into a different cluster. The highest variation among genotypes was observed in the species group (18%), while modern cultivars had the slightest variation among genotypes (1%). The putative origin of early hybrids was analyzed using a likelihood heatmap of all genotypes. Results show what species might be used in breeding for early hybrids. Several modern diploid and tetraploid daylily cultivars have triploid species as ancestors.

Unravelling the genetic diversity and population structure of common walnut in the Iranian Plateau

BackgroundCommon walnut (Juglans regia L.) has a long cultivation history, given its highly valuable wood and rich nutritious nuts. The Iranian Plateau has been considered as one of the last glaciation refugia and a centre of origin and domestication for the common walnut. However, a prerequisite to conserve or utilize the genetic resources of J. regia in the plateau is a comprehensive evaluation of the genetic diversity that is conspicuously lacking. In this regard, we used 31 polymorphic simple sequence repeat (SSR) markers to delineate the genetic variation and population structure of 508 J. regia individuals among 27 populations from the Iranian Plateau.ResultsThe SSR markers expressed a high level of genetic diversity (HO = 0.438, and HE = 0.437). Genetic differentiation among the populations was moderate (FST = 0.124), and genetic variation within the populations (79%) significantly surpassed among populations (21%). The gene flow (Nm = 1.840) may have remarkably influenced the population genetic structure of J. regia, which can be attributed to anthropological activities and wind dispersal of pollen. The STRUCTURE analysis divided the 27 populations into two main clusters. Comparing the neighbor-joining and principal coordinate analysis dendrograms and the Bayesian STRUCTURE analysis revealed the general agreement between the population subdivisions and the genetic relationships among the populations. However, a few geographically close populations dispersed into different clusters. Further, the low genetic diversity of the Sulaymaniyah (SMR) population of Iraq necessitates urgent conservation by propagation and seedling management or tissue culture methods; additionally, we recommend the indispensable preservation of the Gonabad (RGR) and Arak (AKR) populations in Iran.ConclusionsThese results reflected consistent high geographical affinity of the accession across the plateau. Our findings suggest that gene flow is a driving factor influencing the genetic structure of J. regia populations, whereas ecological and geological variables did not act as strong barriers. Moreover, the data reported herein provide new insights into the population structure of J. regia germplasm, which will help conserve genetic resources for the future, hence improving walnut breeding programs’ efficiency.

Genetic Diversity and Association Mapping of Grain-Size Traits in Rice Landraces from the Honghe Hani Rice Terraces System in Yunnan Province.

The Honghe Hani Rice Terraces System (HHRTS) of Yunnan Province is an important agricultural and cultural heritage landscape. Until now, a large number of local rice landraces have been planted. Mining excellent genes contained in these landraces provides a reference for variety improvement and new variety breeding. In this study, 96 rice landraces collected from the Hani terraces were planted in Honghe Mengzi, Yunnan Province, in 2013, 2014, 2015, and 2021, and five major grain traits were measured and analyzed. The genomic variation of 96 rice landraces was scanned by 201 simple sequence repeat (SSR) markers. The genetic diversity, population structure, and genetic relationships of the natural population were analyzed. The mixed linear model (MLM) method of the TASSEL software was used to analyze the associations between markers and traits. A total of 936 alleles were amplified by 201 pairs of SSR primers. The average number of observed alleles (Na), the effective number of alleles (Ne), Shannon's information index (I), heterozygosity (H), and the polymorphism information content (PIC) per marker were 4.66, 2.71, 1.08, 0.15, and 0.55, respectively. Ninety-six landraces were divided into two groups by population structure, clustering, and principal component analysis, and indica rice was the main group. The coefficients of variation of the five traits ranged from 6.80 to 15.24%, and their broad heritabilities were more than 70%. In addition, there were positive correlations among the same grain traits between different years. Through MLM analysis, 2, 36, 7, 7, and 4 SSR markers were significantly associated with grain length (GL), grain width (GW), grain thickness (GT), grain length-width ratio (LWR), and thousand-grain weight (TGW), respectively. The explanation rates of phenotypic variation were 16.31 (RM449, Chr. 1)-23.51% (RM316, Chr. 9), 10.84 (RM523, Chr. 3; RM161/RM305, Chr. 5)-43.01% (RM5496, Chr. 1), 11.98 (RM161/RM305, Chr. 5)-24.72% (RM275, Chr. 6), 12.68 (RM126, Chr. 8)-36.96% (RM5496, Chr. 1), and 17.65 (RM4499, Chr. 2)-26.32% (RM25, Chr. 8), respectively. The associated markers were distributed on 12 chromosomes of the genome.

Creation of New Oregano Genotypes with Different Terpene Chemotypes via Inter- and Intraspecific Hybridization.

Oregano is a medicinal and aromatic plant of value in the pharmaceutical, food, feed additive, and cosmetic industries. Oregano breeding is still in its infancy compared with traditional crops. In this study, we evaluated the phenotypes of 12 oregano genotypes and generated F1 progenies by hybridization. The density of leaf glandular secretory trichomes and the essential oil yield in the 12 oregano genotypes varied from 97-1017 per cm2 and 0.17-1.67%, respectively. These genotypes were divided into four terpene chemotypes: carvacrol-, thymol-, germacrene D/β-caryophyllene-, and linalool/β-ocimene-type. Based on phenotypic data and considering terpene chemotypes as the main breeding goal, six oregano hybrid combinations were performed. Simple sequence repeat (SSR) markers were developed based on unpublished whole-genome sequencing data of Origanum vulgare, and 64 codominant SSR primers were screened on the parents of the six oregano combinations. These codominant primers were used to determine the authenticity of 40 F1 lines, and 37 true hybrids were identified. These 37 F1 lines were divided into six terpene chemotypes: sabinene-, β-ocimene-, γ-terpinene-, thymol-, carvacrol-, and p-cymene-type, four of which (sabinene-, β-ocimene-, γ-terpinene-, and p-cymene-type) were novel (i.e., different from the chemotypes of parents). The terpene contents of 18 of the 37 F1 lines were higher than those of their parents. The above results lay a strong foundation for the creating of new germplasm resources, constructing of genetic linkage map, and mapping quantitative trait loci (QTLs) of key horticultural traits, and provide insights into the mechanism of terpenoid biosynthesis in oregano.

Genetic diversity and the population structure of Monochoria korsakowii in Japan revealed by nuclear simple sequence repeat (SSR) markers

Monochoria korsakowii is an endangered annual herbaceous plant that grows in and around Japanese paddy fields. In the present study, the genetic diversity and population structure were analyzed in 10 populations of M. korsakowii consisting of 200 individuals from Hokkaido, Honshu, and Kyushu, using 15 pairs of nuclear simple sequence repeat (SSR) markers. The results showed that M. korsakowii populations have low genetic diversity (P = 32.9, Ne = 1.0949, I = 0.0911, H = 0.0584, and PPL = 20.56 %), which could be attributed to the highly disturbed habitat, low gene flow, and repeated genetic bottlenecks. The M. korsakowii populations exhibited high genetic differentiation (ranging from 0.5674 to 0.933) owing to the limited gene flow between them. It was inferred that the gene flow between the M. korsakowii populations was hindered by factors such as geographical distance, mountains, straits, and human activities. Thus, comprehensive conservation strategies should be implemented to protect the existing gene pools and enhance population structure.

Molecular and phenotypic markers for pyramiding multiple traits in rice

Abstract Gene pyramiding has been developed for effective resistant genes, such as bacterial blight resistance genes. However, in this report, pyramiding for multiple traits is presented. We evaluated 28 pyramided rice lines derived from 7 hybridizations of valuable parents for high yield potential, early maturity, aroma, high grain quality, and resistance to brown planthopper using both simple-sequence repeat markers and phenotypic markers. Pyramided rice genotypes were evaluated with RM282 molecular marker linked to GW3 gene associated with grain weight, RM259 (number of panicles), RM3600 (length of panicles), RM3701 (short panicles), RM19414 (Hd3), RM7601 (Hd2), RM190, and SSIIa (waxy), IFAP and ESP (aroma), RM586 (Bph3 and Bph4), and RM8213 (Qpbh4 and Bph17(t)). Eight genotypes (i.e., #9, #15, #16, #22, #25, #26, #27, and #28) were confirmed to have desired genes associated with multiple important traits based on molecular markers and morpho-agronomic traits. A strong correlation was found between RM19414 and heading date, between specific markers for aroma and sensory test. Meanwhile, a weak correlation was found between RM259 with panicle number, RM586, and RM8213 with trichome density. As presented in this study, gene pyramiding promises a new hope to combine multiple traits in rice breeding. Promising rice lines will be incorporated in the multiple traits breeding program.

Relocation of Sr48 to Chromosome 2D Using an Alternative Mapping Population and Development of a Closely Linked Marker Using Diverse Molecular Technologies.

The Ug99-effective stem rust resistance gene Sr48 was mapped to chromosome 2A based on its repulsion linkage with Yr1 in an Arina/Forno recombinant inbred line (RIL) population. Attempts to identify markers closely linked to Sr48 using available genomic resources were futile. This study used an Arina/Cezanne F5:7 RIL population to identify markers closely linked with Sr48. Using the Arina/Cezanne DArTseq map, Sr48 was mapped on the short arm of chromosome 2D and it co-segregated with 12 markers. These DArTseq marker sequences were used for BlastN search to identify corresponding wheat chromosome survey sequence (CSS) contigs, and PCR-based markers were developed. Two simple sequence repeat (SSR) markers, sun590 and sun592, and two Kompetitive Allele-Specific PCR (KASP) markers were derived from the contig 2DS_5324961 that mapped distal to Sr48. Molecular cytogenetic analysis using sequential fluorescent in situ hybridization (FISH) and genomic in situ hybridization (GISH) identified a terminal translocation of chromosome 2A in chromosome 2DL of Forno. This translocation would have led to the formation of a quadrivalent involving chromosomes 2A and 2D in the Arina/Forno population, which would have exhibited pseudo-linkage between Sr48 and Yr1 in chromosome 2AL. Polymorphism of the closet marker sunKASP_239 among a set of 178 wheat genotypes suggested that this marker can be used for marker-assisted selection of Sr48.

Identification and genetic mapping of novel resistance gene, Rvn4, for pear scab in Chinese pear

Pear scab, caused by Venturia nashicola, is one of the most harmful diseases of pears, especially of Japanese and Chinese pears. The breeding of new cultivars resistant to pear scab has long been an aim of the Asian pear breeding programs. Although loci for pear scab disease resistance have been identified in Japanese and European pears, information on the genes controlling scab resistance in Chinese pears is limited. Genetic control of scab resistance in Chinese pear is reported to involve a single dominant gene. Here, we reaffirmed the scab resistance in the Chinese pear ‘Hong Li’ controls a single dominant gene and identified the exact location of a scab resistance gene, Rvn4. We first mapped Rvn4 at the top of linkage group (LG) 7 and then used public genome sequences to develop novel simple sequence repeat (SSR) markers to enrich the target region. Using these novel SSRs and the previously published SSR markers, we constructed a LG 7 of ‘Hong Li’ with an average marker density of 1.4 cM per marker. The novel SSR marker Pbr.chr07.20 had a significant linkage to Rvn4, with a genetic distance of 1.3 cM. The identification of the precise locus of Rvn4 in Chinese pear, together with previous studies in Japanese and European pear, will enhance our understanding of the mechanisms of pear scab resistance. DNA markers linked to the novel gene may also allow efficient selection of scab-resistant seedlings in pear breeding programs and the pyramiding of multiple resistance genes.

Complete Genome of Rose Myrtle, Rhodomyrtus tomentosa, and Its Population Genetics in Thai Peninsula.

Several parts of rose myrtle, Rhodomyrtus tomentosa, exhibited profound antibacterial and anti-inflammatory activities, suggesting its potential in healthcare and cosmetics applications. During the past few years, the demand for biologically active compounds in the industrial sectors increased. Therefore, gathering comprehensive information on all aspects of this plant species is essential. Here, the genome sequencing using short and long reads was used to understand the genome biology of R. tomentosa. Inter-simple sequence repeats (ISSR) and simple sequence repeats (SSR) markers, and geometric morphometrics of the leaves of R. tomentosa collected across Thai Peninsula, were determined for population differentiation analysis. The genome size of R. tomentosa was 442 Mb, and the divergence time between R. tomentosa and Rhodamnia argentea, the white myrtle of eastern Australia, was around 15 million years. No population structure was observed between R. tomentosa on the eastern and western sides of the Thai Peninsula using the ISSR and SSR markers. However, significant differences in leaf size and shape of R. tomentosa were observed in all locations.

Microsporogenesis in the triploid hybrid ‘Beilinxiongzhu 1#’ and detection of primary trisomy in 2x × 3 × Populus hybrids

BackgroundPrimary trisomy is a powerful genetic tool in plants. However, trisomy has not been detected in Populus as a model system for tree and woody perennial plant biology.ResultsIn the present study, a backcross between Populus alba × Populus glandulosa ‘YXY 7#’ (2n = 2x = 38) and the triploid hybrid ‘Beilinxiongzhu 1#’ (2n = 3x = 57) based on the observation of microsporogenesis and an evaluation of the variations in pollen was conducted to create primary trisomy. Many abnormalities, such as premature migration of chromosomes, lagging of chromosomes, chromosome bridges, asymmetric separation, micronuclei, and premature cytokinesis, have been detected during meiosis of the triploid hybrid clone ‘Beilinxiongzhu 1#’. However, these abnormal behaviors did not result in completely aborted pollen. The pollen diameter of the triploid hybrid clone ‘Beilinxiongzhu 1#’ is bimodally distributed, which was similar to the chromosomal number of the backcross progeny. A total of 393 progeny were generated. We provide a protocol for determining the number of chromosomes in aneuploid progeny, and 19 distinct simple sequence repeat (SSR) primer pairs covering the entire Populus genome were developed. Primary trisomy 11 and trisomy 17 were detected in the 2x × 3 x hybrid using the SSR molecular markers and counting of somatic chromosomes.ConclusionsNineteen distinct SSR primer pairs for determining chromosomal number in aneuploid individuals were developed, and two Populus trisomies were detected from 2x × 3 x hybrids by SSR markers and somatic chromosome counting. Our findings provide a powerful genetic tool to reveal the function of genes in Populus.

Novel hybrids with floral scent between Lagerstroemia caudata and three L. indica cultivars

L. caudata, native to China, is valued for its floral fragrance while flower size and color still negatively affects their economic value. In this study, three L. indica cultivars ‘Bai Yun Ying Xia’, ‘Qiao Jia Ren’ and ‘Fen Hu Die’ with diverse flower traits were chosen to make cross with L. caudata. Finally, a total of 2832 crosses were obtained, including interspecific, backcross and self-pollination combinations between L. caudata and L. indica. Nearly 260 hybrid seedlings were survived. Hybridity was confirmed by both morphological comparison and simple sequence repeat markers (SSR) technique. All obtained interspecific seedlings were generally intermediate in morphological traits of their parents, but variable in leaf blade length, leaf blade width, leaf blade length/width ratio, flower diameter, flower claw length, petal length, flower color and floral fragrance. Our results suggested that it is possible to optimize floral fragrance of Lagerstroemia cultivars by using L. caudata and the hybrids obtained in this study. These findings could be served as the basis for the development of superior plants with unique combinations of ornamental traits.

Distribution of genetic variability in mature and progeny populations of Abies alba Mill. from the Polish Western and Eastern Carpathians

This study aimed to compare patterns in the genetic structure of 27 mature stands and the natural regeneration of A. alba in the Eastern and Western Carpathians within the introgression zone of two refugial lineages from the Apennine and Balkan peninsulas. The distribution of the genetic diversity of fir stands was analysed using paternally inherited chloroplast DNA and five nuclear dominant inter simple sequence repeat markers (ISSRs). The study showed that the Balkan haplotype prevailed in both parental and progeny populations, and this haplotype was found in many mature Eastern Carpathian stands. Relatively high levels of genetic diversity were found in the mature stands (effective number of alleles Ne = 1.517, Shannon index I = 0.436, expected heterozygosity He = 0.295) and progeny (Ne = 1.515, I = 0.436 and He = 0.294) of silver fir. The analysis of molecular variance (AMOVA) revealed slight differences among the mature fir stands from the Western and Eastern Carpathians, with a value of 1.1%. According to principal coordinates analysis (PCoA) and STRUCTURE analyses, the populations, including stands of mature and progeny trees, were genetically separated into two groups. Slight genetic differences between the mature and progeny populations in the Polish Carpathians indicate sound gene pool transmission, which is essential for creating new selection and breeding programmes.

SSR and SNP Marker-Based Investigation of Indian Rice Landraces in Relation to Their Genetic Diversity, Population Structure, and Geographical Isolation

India is blessed with an abundance of diverse rice landraces in its traditional cultivated areas. Two marker systems (simple sequence repeats (SSR) and single nucleotide polymorphism (SNP)) were used to study a set of 298 rice landrace accessions collected from six different regions of India (Andaman and Nicobar Islands, Chhattisgarh, Jharkhand, Uttar Pradesh, Uttarakhand, and West Bengal). Thirty hyper-variable simple sequence repeats (HvSSRs) and 32,782 single nucleotide polymorphisms (SNPs) were used in inferring genetic structure and geographical isolation. Rice landraces from Uttar Pradesh were the most diverse, with a gene diversity value of 0.42 and 0.49 with SSR and SNP markers, respectively. Neighbor-joining trees classified the rice landraces into two major groups with SSR and SNP markers, and complete geographical isolation was observed with SSR markers. Fast STRUCTURE analysis revealed four populations for SSR markers and three populations for SNP markers. The population structure with SSR markers showed that few individuals from Uttarakhand and Andaman and Nicobar Islands were grouped in small clusters. Population structure analysis with SNP markers showed not very distinct region-wise clustering among the rice landraces. Discriminant analysis of principal components (DAPC) and minimum spanning network (MSN) using SSR markers showed region-wise grouping of landraces with some intermixing, but DAPC and MSN with SNP markers showed very clear region-wise clustering. Genetic differentiation of rice landraces between the regions was significant with both SSR (Fst 0.094–0.487) and SNP markers (Fst 0.047–0.285). A Mantel test revealed a positive correlation between the genetic and geographic distance of rice landraces. The present study concludes that rice landraces investigated in this study were very diverse, and unlinked SSR markers show better geographical isolation than a large set of SNP markers.

A set of SSR markers to characterize genetic diversity in all Viburnum species

About 160 species are classified within the Viburnum genus and many of these are cultivated for horticultural purposes. The vast dispersal of Viburnum makes the genus a useful model for studying evolutionary history and inferring how species expanded into their current distributions. Simple sequence repeat (SSR) markers were previously developed for five Viburnum species that were classified within the four major clades (Laminotinus, Crenotinus, Valvatotinus, and Porphyrotinus). The ability of some of these markers to cross-amplify in Viburnum species has been scantly evaluated, but there has not been any genus-wide assessment for the markers. We evaluated a collection of 49 SSR markers for the ability to cross-amplify in 224 samples, including 46 Viburnum species, representing all 16 subclades, and five additional species in the Viburnaceae and Caprifoliaceae. A subset of 14 potentially comprehensive markers for Viburnum species was identified and evaluated for the ability to detect polymorphisms in species outside of their respective clades. The 49 markers had overall amplification success in 52% of the samples, including a 60% success rate within the Viburnum genus and 14% in other genera. The comprehensive marker set amplified alleles in 74% of all samples tested, including 85% of Viburnum samples and 19% of outgroup samples. To the best of our knowledge, this is the first comprehensive set of markers able to characterize species across an entire genus. This set of markers can be used to assess the genetic diversity and population structure of most Viburnum species and closely allied species.