Chinese Bulletin of Botany ›› 2015, Vol. 50 ›› Issue (6): 754-764.DOI: 10.11983/CBB14173

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Validation of Reference Genes for Quantitative RT-PCR Analysis in Soybean Root Tissue under Salt Stress

Qiong Jiang1,2, Youning Wang1, Lixiang Wang1,2, Zhengxi Sun1,2, Xia Li1*   

  1. 1Center of Agricultural Research Resources, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Shijiazhuang 050021, China
    2University of Chinese Academy of Sciences, Beijing 100049, China
  • Received:2014-09-22 Accepted:2015-01-14 Online:2015-11-01 Published:2015-09-06
  • Contact: Li Xia
  • About author:

    ? These authors contributed equally to this paper

Abstract: Quantitative RT-PCR (qRT-PCR) is a robust and commonly used technique to study gene expression. However, the reference genes and microRNA (miRNA) that can be used in soybean molecular research, in particular the stress responsive genes and miRNA in roots, are not well characterized. In this study, we systematically analyzed the expression stability of eight widely used reference genes (ACT, ACT2/7, CYP2, ELF1A, ELF1B, F-Box, TUA and UBC2) and miRNA internal references (U6, miR1515a, miR1520c, miR1520d, miR171a, and miR171b) in root tissues treated with different NaCl concentrations at specified times. The Δ-Ct method, Bestkeeper 1.0, NormFinder 0.953 and Genorm 3.5 were used to analyze the data, then the web-based tool RefFinder was used to integrate the results obtained. ELF1B/CYP2 and miR1515a/U6 were the most suitable genes for qRT-PCR analysis of soybean gene and miRNA expression analysis under salt stress, respectively. The highly ranked reference genes and miRNAs identified from this study can be used for detecting differences in expression rates of genes and miRNAs in response to salt stress in soybean.