Development and Validation of EST-SSR Markers from the Transcriptome of Adzuki Bean (Vigna angularis)

PLoS One. 2015 Jul 6;10(7):e0131939. doi: 10.1371/journal.pone.0131939. eCollection 2015.

Abstract

The adzuki bean (Vigna angularis (Ohwi) Ohwi and Ohashi) is an important grain legume of Asia. It is cultivated mainly in China, Japan and Korea. Despite its importance, few genomic resources are available for molecular genetic research of adzuki bean. In this study, we developed EST-SSR markers for the adzuki bean through next-generation sequencing. More than 112 million high-quality cDNA sequence reads were obtained from adzuki bean using Illumina paired-end sequencing technology, and the sequences were de novo assembled into 65,950 unigenes. The average length of the unigenes was 1,213 bp. Among the unigenes, 14,547 sequences contained a unique simple sequence repeat (SSR) and 3,350 sequences contained more than one SSR. A total of 7,947 EST-SSRs were identified as potential molecular markers, with mono-nucleotide A/T repeats (99.0%) as the most abundant motif class, followed by AG/CT (68.4%), AAG/CTT (30.0%), AAAG/CTTT (26.2%), AAAAG/CTTTT (16.1%), and AACGGG/CCCGTT (6.0%). A total of 500 SSR markers were randomly selected for validation, of which 296 markers produced reproducible amplicons with 38 polymorphic markers among the 32 adzuki bean genotypes selected from diverse geographical locations across China. The large number of SSR-containing sequences and EST-SSR markers will be valuable for genetic analysis of the adzuki bean and related Vigna species.

Publication types

  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Expressed Sequence Tags*
  • Fabaceae / genetics*
  • Genes, Plant*
  • Genetic Markers
  • Genotype
  • High-Throughput Nucleotide Sequencing*
  • Microsatellite Repeats*
  • Transcriptome*

Substances

  • Genetic Markers

Associated data

  • SRA/SRP049807

Grants and funding

This study was funded by Agricultural Science and Technology Innovation Program (ASTIP) in CAAS and China Agriculture Research System (CARS-09) from the Ministry of Agriculture of China.