Distribution and analysis of SSR in mung bean (Vigna radiata L.) genome based on an SSR-enriched library

Abstract Simple sequence repeats (SSR) are widely distributed in plant genomes, have been popular genetic markers and can be involved in gene function. We report an SSR analysis of mung bean (Vigna radiata L.), based on an SSR-enriched library. A total of 308,509 SSRs (56.9 % simple and 43.1 % compound) were discovered from 167,628 sequences. Both di- and tri-nucleotide were the most prevalent repeat types (each accounts for 48.6 %). The most frequent motifs were AAC/GTT, accounting for 45.14 % of all, followed by AC/GT at 41.80 %. After filtering the SSRs, 70,104 flanking sequences were used as BLAST queries and corresponded to 574 non-redundant gene ontology terms against the protein database from Arabidopsis thaliana. The three main categories were biological processes (23.8 %), cellular components (44.4 %) and molecular functions (31.8 %). A total of 6,100 non-repeated primer pairs were designed and validated by PCR analysis. The results showed that 60 % of primers were effective in mung bean and 35.2, 34.0 and 25.9 % could be transferred to rice bean, adzuki bean and cowpea, respectively. 9.1 % of the 6,100 displayed polymorphism between a wild and cultivated mung bean genotype, and 367 were mapped onto chromosomes using a RIL population derived from a wild × a cultivated cross. However, only 49 of 1,700 effective primer pairs showed polymorphism among 32 Chinese cultivated mung bean accessions. A total of 46,565 loci on mung bean chromosomes from the draft genome were hit by the 70,104 flanking sequences using BLASTn. The present study, especially the newly published 387 markers that have been validated and mapped, will significantly enhance genetic linkage map construction, QTL mapping and marker-assisted selection in mung bean and breeding in closely related crop species.

[1]  H. Enoki,et al.  SSR analysis of genetic diversity among maize inbred lines adapted to cold regions of Japan , 2002, Theoretical and Applied Genetics.

[2]  Rajeev K. Varshney,et al.  Genome sequence of mungbean and insights into evolution within Vigna species , 2014, Nature Communications.

[3]  M. Karaca,et al.  Simple sequence repeat (SSR) markers linked to the Ligon lintless (Li(1)) mutant in cotton. , 2002, The Journal of heredity.

[4]  Ashok Sharma,et al.  Analysis of Unigene Derived Microsatellite Markers in Family Solanaceae , 2010 .

[5]  J. Manners,et al.  Two genetic linkage maps of mungbean using RFLP and RAPD markers , 2000 .

[6]  Arwa Shahin,et al.  Generation and analysis of expressed sequence tags in the extreme large genomes Lilium and Tulipa , 2012, BMC Genomics.

[7]  Yong-Jin Park,et al.  Characterization of new microsatellite markers in mung bean, Vigna radiata (L.) , 2006 .

[8]  T. Magner,et al.  Development of a mungbean (Vigna radiata) RFLP linkage map and its comparison with lablab (Lablab purpureus) reveals a high level of colinearity between the two genomes , 2002, Theoretical and Applied Genetics.

[9]  Sarika Gupta,et al.  Development and characterization of genic SSR markers in Medicago truncatula and their transferability in leguminous and non-leguminous species. , 2009, Genome.

[10]  Steven G. Schroeder,et al.  Development and mapping of SSR markers for maize , 2002, Plant Molecular Biology.

[11]  N. Tomooka,et al.  Genetic diversity of the azuki bean (Vigna angularis (Willd.) Ohwi & Ohashi) gene pool as assessed by SSR markers. , 2008, Genome.

[12]  Yong-Jin Park,et al.  Sequence information on simple sequence repeats and single nucleotide polymorphisms through transcriptome analysis of mungbean. , 2011, Journal of integrative plant biology.

[13]  S. Tabata,et al.  Construction of a Genetic Linkage Map and Genetic Analysis of Domestication Related Traits in Mungbean (Vigna radiata) , 2012, PloS one.

[14]  M. Morgante,et al.  PCR-amplified microsatellites as markers in plant genetics. , 1993, The Plant journal : for cell and molecular biology.

[15]  L. Qiu,et al.  Genetic Diversity of Chinese Cultivated Soybean Revealed by SSR Markers , 2006 .

[16]  Juan Miguel García-Gómez,et al.  Sequence analysis Blast 2 GO : a universal tool for annotation , visualization and analysis in functional genomics research , 2005 .

[17]  D. J. Perry Identification of Canadian durum wheat varieties using a single PCR , 2004, Theoretical and Applied Genetics.

[18]  P. Somta,et al.  Genetic diversity of the Bambara groundnut (Vigna subterranea (L.) Verdc.) as assessed by SSR markers. , 2011, Genome.

[19]  S. Gupta,et al.  Development and characterization of genic SSR markers for mungbean (Vigna radiata (L.) Wilczek) , 2013, Euphytica.

[20]  N. Tomooka,et al.  Development of a black gram [Vigna mungo (L.) Hepper] linkage map and its comparison with an azuki bean [Vigna angularis (Willd.) Ohwi and Ohashi] linkage map , 2006, Theoretical and Applied Genetics.

[21]  N. Tomooka,et al.  The development of SSR markers by a new method in plants and their application to gene flow studies in azuki bean [Vigna angularis (Willd.) Ohwi & Ohashi] , 2004, Theoretical and Applied Genetics.

[22]  Florian Martin,et al.  An application of kernel methods to variety identification based on SSR markers genetic fingerprinting , 2011, BMC Bioinformatics.

[23]  E. Pahlich,et al.  A rapid DNA isolation procedure for small quantities of fresh leaf tissue , 1980 .

[24]  R. Deshmukh,et al.  Genome-Wide Distribution and Organization of Microsatellites in Plants: An Insight into Marker Development in Brachypodium , 2011, PloS one.

[25]  Wang Li Genetic Diversity of Adzuki Bean Germplasm Resources Revealed by SSR Markers , 2009 .

[26]  M. Mimura,et al.  RAPD variation in wild, weedy and cultivated azuki beans in Asia , 2000, Genetic Resources and Crop Evolution.

[27]  T. Harkins,et al.  Genome-wide characterization of simple sequence repeats in cucumber (Cucumis sativus L.) , 2010, BMC Genomics.

[28]  N. Tomooka,et al.  Genetic diversity of the rice bean (Vigna umbellata) genepool as assessed by SSR markers. , 2013, Genome.

[29]  S. Weissman,et al.  Construction of libraries enriched for sequence repeats and jumping clones, and hybridization selection for region-specific markers. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[30]  R. Varshney,et al.  Exploiting EST databases for the development and characterization of gene-derived SSR-markers in barley (Hordeum vulgare L.) , 2003, Theoretical and Applied Genetics.

[31]  Nigel Maxted,et al.  The Asian Vigna:: Genus Vigna subgenus Ceratotropis Genetic Resources , 2003 .

[32]  Shanmughavel Piramanayagam,et al.  Mining of SSR markers from Expressed Sequence Tags of bamboo species , 2010, Bioinformation.

[33]  M. Badenes,et al.  Development and characterization of microsatellite markers in pomegranate (Punica granatum L.) , 2010, Molecular Breeding.

[34]  P. Somta,et al.  Development, characterization and cross-species amplification of mungbean (Vigna radiata) genic microsatellite markers , 2009, Conservation Genetics.

[35]  Somvong Tragoonrung,et al.  Characterization of microsatellites and gene contents from genome shotgun sequences of mungbean (Vigna radiata (L.) Wilczek) , 2009, BMC Plant Biology.

[36]  P. Somta,et al.  Molecular diversity assessment of AVRDC–The World Vegetable Center elite-parental mungbeans , 2009 .

[37]  J. Jurka,et al.  Microsatellites in different eukaryotic genomes: survey and analysis. , 2000, Genome research.

[38]  A. Myburg,et al.  Genetic diversity of African maize inbred lines revealed by SSR markers. , 2007, Hereditas.

[39]  K. Yusoff,et al.  Isolation and characterization of seven tetranucleotide microsatellite loci in mungbean, Vigna radiata , 2002 .

[40]  Liqing Zhang,et al.  Distinct patterns of SSR distribution in the Arabidopsis thaliana and rice genomes , 2006, Genome Biology.

[41]  F. D. de Carvalho,et al.  Tandem repeat distribution of gene transcripts in three plant families , 2009, Genetics and molecular biology.

[42]  A. F. Robinson,et al.  Identification of QTL regions and SSR markers associated with resistance to reniform nematode in Gossypium barbadense L. accession GB713 , 2011, Theoretical and Applied Genetics.

[43]  Xuzhen Cheng,et al.  Construction of Mungbean Genetic Linkage Map , 2010 .

[44]  T. Yang,et al.  High-throughput novel microsatellite marker of faba bean via next generation sequencing , 2012, BMC Genomics.

[45]  O. Lepais,et al.  Comparison of random and SSR‐enriched shotgun pyrosequencing for microsatellite discovery and single multiplex PCR optimization in Acacia harpophylla F. Muell. Ex Benth , 2011, Molecular ecology resources.

[46]  S Rozen,et al.  Primer3 on the WWW for general users and for biologist programmers. , 2000, Methods in molecular biology.

[47]  N. Tomooka,et al.  Genetic diversity of the mungbean (Vigna radiata, Leguminosae) genepool on the basis of microsatellite analysis , 2007 .

[48]  C. P. Hong,et al.  Genomic distribution of simple sequence repeats in Brassica rapa. , 2007, Molecules and cells.

[49]  L. Ya Genetic Diversity Research of Mungbean Germplasm Resources by SSR Markers in China , 2013 .

[50]  R. Chibbar,et al.  Development of microsatellite markers in canary seed (Phalaris canariensis L.) , 2011, Molecular Breeding.

[51]  D. Poudel,et al.  Identification of SSR markers associated with saccharification yield using pool-based genome-wide association mapping in sorghum. , 2011, Genome.

[52]  P. Somta,et al.  Quantitative trait loci mapping of Cercospora leaf spot resistance in mungbean, Vigna radiata (L.) Wilczek , 2011, Molecular Breeding.

[53]  N. Tomooka,et al.  A genetic linkage map for azuki bean [Vigna angularis (Willd.) Ohwi & Ohashi] , 2005, Theoretical and Applied Genetics.

[54]  W. Suhua,et al.  Genetic Diversity of Adzuki Bean Germplasm Resources Revealed by SSR Markers , 2009 .

[55]  E. Nevo,et al.  Microsatellites: genomic distribution, putative functions and mutational mechanisms: a review , 2002, Molecular ecology.

[56]  M. Bateson,et al.  Construction of bacterial artificial chromosome libraries and their application in developing PCR-based markers closely linked to a major locus conditioning bruchid resistance in mungbean (Vigna radiata L. Wilczek) , 2004, Theoretical and Applied Genetics.

[57]  D. Berry,et al.  Assessing probability of ancestry using simple sequence repeat profiles: applications to maize hybrids and inbreds. , 2002, Genetics.

[58]  M. Wingfield,et al.  Microsatellite discovery by deep sequencing of enriched genomic libraries. , 2009, BioTechniques.

[59]  C. Liang,et al.  Genome-Wide Analysis of Tandem Repeats in Plants and Green Algae , 2013, G3: Genes, Genomes, Genetics.

[60]  张静,et al.  Banana Ovate family protein MaOFP1 and MADS-box protein MuMADS1 antagonistically regulated banana fruit ripening , 2015 .

[61]  R. Wing,et al.  Gene-based SSR markers for common bean (Phaseolus vulgaris L.) derived from root and leaf tissue ESTs: an integration of the BMc series , 2011, BMC Plant Biology.

[62]  Li Changyou,et al.  Transferability of SSR from adzuki bean to mungbean. , 2009 .

[63]  R. Sundaram,et al.  Identification of flanking SSR markers for a major rice gall midge resistance gene Gm1 and their validation , 2004, Theoretical and Applied Genetics.

[64]  G. Cutting,et al.  A variable dinucleotide repeat in the CFTR gene contributes to phenotype diversity by forming RNA secondary structures that alter splicing. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[65]  S. Korban,et al.  Identification, characterization, and utilization of genome-wide simple sequence repeats to identify a QTL for acidity in apple , 2012, BMC Genomics.

[66]  Y. Kashi,et al.  Simple sequence repeats as a source of quantitative genetic variation. , 1997, Trends in genetics : TIG.

[67]  G. Scoles,et al.  Determining Genetic Similarities and Relationships among Cowpea Breeding Lines and Cultivars by Microsatellite Markers , 2001 .

[68]  L. Lipovich,et al.  Computational and experimental analysis of microsatellites in rice (Oryza sativa L.): frequency, length variation, transposon associations, and genetic marker potential. , 2001, Genome research.

[69]  T. Toojinda,et al.  New microsatellite markers isolated from mungbean (Vigna radiata (L.) Wilczek) , 2008, Molecular ecology resources.

[70]  Changyou Liu,et al.  Transferability of SSR from Adzuki Bean to Mungbean: Transferability of SSR from Adzuki Bean to Mungbean , 2009 .

[71]  C. Lian,et al.  An improved technique for isolating codominant compound microsatellite markers , 2006, Journal of Plant Research.

[72]  Chun Liang,et al.  An annotated genetic map of loblolly pine based on microsatellite and cDNA markers , 2011, BMC Genetics.

[73]  S. Tan,et al.  Isolation of microsatellite markers in mungbean, Vigna radiata , 2002 .

[74]  A. Jeffreys,et al.  Isolation of human simple repeat loci by hybridization selection. , 1994, Human molecular genetics.

[75]  Sumana Banerjee,et al.  Development of SSR markers and construction of a linkage map in jute , 2012, Journal of Genetics.

[76]  T. Glenn,et al.  Isolating microsatellite DNA loci. , 2005, Methods in enzymology.

[77]  Cai-rui Lu,et al.  Distribution and characterization of simple sequence repeats in Gossypium raimondii genome , 2012, Bioinformation.

[78]  C. Schlötterer,et al.  Genome evolution: Are microsatellites really simple sequences? , 1998, Current Biology.

[79]  B. Jayashree,et al.  A Database of Simple Sequence Repeats from Cereal and Legume Expressed Sequence Tags Mined in silico: Survey and Evaluation , 2006, Silico Biol..