Reference gene selection for quantitative real-time PCR (qRT-PCR) expression analysis in Galium aparine L

Xu Su; Liuyang Lu; Yashe Li; Congai Zhen; Guilei Hu; Kun Jiang; Yawei Yan; Yanbo Xu; Geng Wang; Mingwang Shi; Xiling Chen; Baizhong Zhang

doi:10.1371/journal.pone.0226668

Reference gene selection for quantitative real-time PCR (qRT-PCR) expression analysis in Galium aparine L

PLoS One. 2020 Feb 4;15(2):e0226668. doi: 10.1371/journal.pone.0226668. eCollection 2020.

Authors

Xu Su¹, Liuyang Lu¹, Yashe Li¹, Congai Zhen², Guilei Hu¹, Kun Jiang¹, Yawei Yan¹, Yanbo Xu¹, Geng Wang¹, Mingwang Shi¹, Xiling Chen¹, Baizhong Zhang¹

Affiliations

¹ College of Resources and Environment, Henan Institute of Science and Technology, Xinxiang, P.R. China.
² The Museum of Chinese Gardens and Landscape Architecture, Beijing, P.R. China.

Abstract

To accurately evaluate expression levels of target genes, stable internal reference genes is required for normalization of quantitative real-time PCR (qRT-PCR) data. However, there have been no systematical investigation on the stability of reference genes used in the bedstraw weed, Galium aparine L. (BGA). In this study, the expression profiles of seven traditionally used reference genes, namely 18S, 28S, ACT, GAPDH, EF1α, RPL7 and TBP in BGA were assessed under both biotic (developmental time and tissue), and abiotic (temperature, regions and herbicide) conditions. Four analytical algorithms (geNorm, Normfinder, BestKeeper and the ΔCt method) were used to analyze the suitability of these genes as internal reference genes. RefFinder, a comprehensive analytical software, was used to rank the overall stability of the candidate genes. The optimal normalization internal control genes were ranked as: 28S and RPL7 were best for all the different experimental conditions (developmental stages, tissues, temperature, regions and herbicide treatment); 28S and RPL7 for developmental stages; TBP and GAPDH for different tissues; 28S and GAPDH were relatively stable for different temperature; 28S and TBP were suitable for herbicide treatment. A specific set of reference genes were recommended for each experimental condition in BGA.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Galium / genetics*
Gene Expression Profiling / methods*
Real-Time Polymerase Chain Reaction / standards*
Reference Standards
Software

Grants and funding

This work is supported by the National Key Research and Development Program of China (2017YFD0201700) to XC, the Key Science and Technology Program of Henan (Agriculture) (182102110053) to BZ, and Project of Plant Protection Key Discipline of Henan Province (1070202190011005). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.