Abstract
Faba bean (Vicia faba L.) is an important food legume crop with a huge genome. Development of genetic markers for faba bean is important to study diversity and for molecular breeding. In this study, we used Next Generation Sequencing (NGS) technology for the development of genomic simple sequence repeat (SSR) markers. A total of 14,027,500 sequence reads were obtained comprising 4,208 Mb. From these reads, 56,063 contigs were assembled (16,367 Mb) and 2138 SSRs were identified. Mono and dinucleotides were the most abundant, accounting for 57.5 % and 20.9 % of all SSR repeats, respectively. A total of 430 primer pairs were designed from contigs larger than 350 nucleotides and 50 primers pairs were tested for validation of SSR locus amplification. Nearly all (96 %) of the markers were found to produce clear amplicons and to be reproducible. Thirty-nine SSR markers were then applied to 46 faba bean accessions from worldwide origins, resulting in 161 alleles with 87.5 % polymorphism, and an average of 4.1 alleles per marker. Gene diversity (GD) of the markers ranged from 0 to 0.48 with an average of 0.27. Testing of the markers showed that they were useful in determining genetic relationships and population structure in faba bean accessions.
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Acknowledgments
This study was supported by grant 0424.STZ.2013-2 from The Republic of Turkey’s Ministry of Science, Industry and Technology to AF with contributions from Polen Seed Co. Additional support was received from the Scientific and Technological Research Council of Turkey grant 113E326 to JA. We are grateful to Ali Uncu and Ayse Ozger Uncu for SSR validation by sequencing.
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ESM 1
Supplementary Table S1 Preprocessing and assembly statistics for the faba bean genomic sequences. Supplementary Table S2 Primer details for the 11 SSR markers not given in Table 3. Supplementary Table S3 Faba bean genomic sequence displaying the four simple sequence repeat motifs within the PCR amplicons. Supplementary Fig. S1 Bar plot showing genetic structure of 46 faba bean accessions with K = 2. Each accession is represented by a vertical bar which is partitioned according to estimated membership in the two clusters. Cluster A fraction is shown in red and cluster B fraction in green. (DOCX 123 kb)
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Abuzayed, M.A., Goktay, M., Allmer, J. et al. Development of genomic simple sequence repeat markers in faba bean by next-generation sequencing. Plant Mol Biol Rep 35, 61–71 (2017). https://doi.org/10.1007/s11105-016-1003-1
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DOI: https://doi.org/10.1007/s11105-016-1003-1