Identification of reliable reference genes for quantitative real-time PCR analysis of the Rhus chinensis Mill. leaf response to temperature changes,FEBS Open Bio

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Identification of reliable reference genes for quantitative real-time PCR analysis of the Rhus chinensis Mill. leaf response to temperature changes
FEBS Open Bio ( IF 2.8 ) Pub Date : 2021-08-17 , DOI: 10.1002/2211-5463.13275
Yanchao Chen _{1,

2} , Biao Luo _{1,

2} , Chuwei Liu _{1,

2} , Zhengfeng Zhang ₁ , Chi Zhou _{1,

2} , Ting Zhou _{1,

2} , Guoping Peng _{1,

2} , Xujun Wang ₃ , Waichin Li ₄ , Chuan Wu _{4,

5} , Liqun Rao _{1,

2} , Qiming Wang _{1,

2}

Affiliation

Rhus chinensis Mill. (RCM) is the host plant of Galla chinensis, which is valued in traditional medicine. Environmental temperature directly determines the probability of gallnut formation and RCM growth. At present, there is no experiment to systematically analyse the stability of internal reference gene (RG) expression in RCM. In this experiment, leaves that did not form gallnuts were used as the control group, while leaves that formed gallnuts were used as the experimental group. First, we conducted transcriptome experiments on RCM leaves to obtain 45 103 differential genes and functional enrichment annotations between the two groups. On this basis, this experiment established a transcriptional gene change model of leaves in the process of gallnut formation after being bitten by aphids, and RCM reference candidate genes were screened from RNA sequencing (RNA-seq) data. This study is based on RCM transcriptome data and evaluates the stability of 11 potential reference genes under cold stress (4 °C) and heat stress (34 °C), using three statistical algorithms (geNorm, NormFinder, and BestKeeper). The results show that GAPDH1 + PP2A2/UBQ are stable reference genes under heat stress, while GAPDH1 + ACT are the most stable under cold stress. This study is the first to screen candidate reference genes in RCM and could help guide future molecular studies in this genus.

中文翻译：

鉴定用于Rhus chinensis Mill定量实时PCR分析的可靠参考基因。叶对温度变化的反应

漆树磨坊。(RCM) 是五倍子的寄主植物，这在传统医学中很有价值。环境温度直接决定五倍子形成和 RCM 生长的概率。目前还没有实验系统分析RCM中内参基因（RG）表达的稳定性。在本实验中，未形成五倍子的叶子作为对照组，而形成五倍子的叶子作为实验组。首先，我们对 RCM 叶片进行了转录组实验，获得了 45 103 个差异基因和两组之间的功能富集注释。在此基础上，本实验建立了叶片在被蚜虫叮咬后五倍子形成过程中的转录基因变化模型，并从RNA测序（RNA-seq）数据中筛选出RCM参考候选基因。本研究基于 RCM 转录组数据，使用三种统计算法（geNorm、NormFinder 和 BestKeeper）评估了 11 个潜在参考基因在冷应激（4°C）和热应激（34°C）下的稳定性。结果表明GAPDH1 + PP2A2/UBQ是热胁迫下稳定的参考基因，而GAPDH1 + ACT在冷胁迫下是最稳定的。这项研究是第一个在 RCM 中筛选候选参考基因的研究，可以帮助指导该属的未来分子研究。

更新日期：2021-10-02

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