Selection and Validation of Reference Genes for Gene Expression Analysis in Vigna angularis Using Quantitative Real-Time RT-PCR

PLoS One. 2016 Dec 16;11(12):e0168479. doi: 10.1371/journal.pone.0168479. eCollection 2016.

Abstract

Adzuki bean (Vigna angularis) is one of the most important legume crops in Asian countries like China, Japan and Korea due to its nutritious protein and starch contents. In spite of its economic importance, gene expression analysis system for gene function verification of adzuki bean is still absent. Therefore, reference genes for gene expression analysis based on the quantitative real time PCR (qRT-PCR) were screened in current study. A total of nine general housekeeping genes, including ACT, Fbox, ZMPP, GAPDH, EF, PP2A, UBC, UBN and PTB were evaluated for their expression stability by qRT-PCR in four adzuki bean cultivars, three different tissues, four abiotic stress and one biotic stress. The best group of candidates as reference genes were as follows: PTB and ACT for different cultivars; EF and UBN for different tissues; ACT and ZMPP for biotic stress and waterlogging stress; Fbox and UBC for salinity-alkalinity stress; Fbox and PTB for drought stress. Our results will provide a more accurate and reliable normalization of qRT-PCR data in adzuki bean.

MeSH terms

  • Crops, Agricultural / genetics
  • Droughts
  • Gene Expression Profiling / standards*
  • Gene Expression Regulation, Plant
  • Genes, Essential*
  • Plant Proteins / genetics*
  • Real-Time Polymerase Chain Reaction / standards
  • Reference Standards
  • Reverse Transcriptase Polymerase Chain Reaction / standards
  • Salinity
  • Stress, Physiological
  • Vigna / genetics*
  • Vigna / growth & development

Substances

  • Plant Proteins

Grants and funding

2014BAD07B05-06/H08 http://www.most.gov.cn/, Ministry of Science and Technology of the People's Republic of China, ZYH received the funding. The funder role in study design and decision to publish. 31501629, https://isisn.nsfc.gov.cn/egrantweb/, National Natural Science Foundation of China, KXW received the funding. The funder role in data collection. 2014M561378, http://jj.chinapostdoctor.org.cn/V1/Program2/Default.aspx, China Postdoctoral Science Foundation, YLH received the funding. The funder role in data analysis.