Validation of reference genes for gene expression studies in tartary buckwheat (Fagopyrum tataricum Gaertn.) using quantitative real-time PCR

Chenglei Li; Haixia Zhao; Maofei Li; Panfeng Yao; Qingqing Li; Xuerong Zhao; Anhu Wang; Hui Chen; Zizhong Tang; Tongliang Bu; Qi Wu

doi:10.7717/peerj.6522

Validation of reference genes for gene expression studies in tartary buckwheat (Fagopyrum tataricum Gaertn.) using quantitative real-time PCR

PeerJ. 2019 Feb 26:7:e6522. doi: 10.7717/peerj.6522. eCollection 2019.

Authors

Chenglei Li¹, Haixia Zhao¹, Maofei Li¹, Panfeng Yao¹, Qingqing Li¹, Xuerong Zhao¹, Anhu Wang², Hui Chen¹, Zizhong Tang¹, Tongliang Bu¹, Qi Wu¹

Affiliations

¹ College of Life Science, Sichuan Agricultural University, Ya'an, Sichuan, China.
² Xichang College, Xichang, Sichuan, China.

Abstract

Quantitative real-time reverse transcriptase polymerase chain reaction is a sensitive technique for quantifying gene expression levels. By implementing three distinct algorithms (geNorm, normFinder and BestKeeper), we have validated the stability of the expression of seven candidate reference genes in tartary buckwheat, including FtSAND, FtCACS, FtExpressed1, FtGAPDH, FtActin, FtEF-1a and FtH3. In this study, the results indicated that FtCACS and FtSAND were the best reference genes for 'abiotic cotyledons', FtExpressed1 and FtEF-1α were the best reference genes for aluminium treatment, FtCACS and FtExpressed1 performed the best for the immature seed stage, FtCACS was best for the abiotic treatment, and FtH3 appeared to be the most suitable reference gene for the abiotic treatment in hypocotyls and all samples in this study. In contrast, FtActin and FtGAPDH are unsuitable genes. Our findings offer additional stable reference genes for gene expression research on tartary buckwheat at the immature seed stage and under abiotic treatment.

Keywords: Quantitative real-time PCR; Reference genes; Tartary buckwheat.

Grants and funding

This work was supported by the National Natural Science Foundation of China (31500289). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.