稻螟赤眼蜂实时荧光定量PCR内参基因的筛选

doi:10.16409/j.cnki.2095-039x.2021.11.004

摘要/Abstract

摘要： 为了筛选适合稻螟赤眼蜂试验使用的内参基因，本试验根据转录组测序结果挑选了十个稻螟赤眼蜂候选内参基因（包括PABPC1L、ACTR2、ARPC2、EF1a、EEF1D、SFRS7、HSP70、CSNK1a1、GAPDH和TMEM209），分别检测了其在包括温度、取食、农药刺激等不同条件下，以及在稻螟赤眼蜂不同的发育历期的表达情况。利用RefFinder分析软件对Delta CT、BestKeeper、NormFinder、GeNorm四种软件分析基因表达稳定性的结果进行综合排序。结果表明，不同温度、取食、农药刺激和不同发育历期处理，表达相对稳定的内参基因分别为EF1a、EEF1D；SFRS7、ARPC2；PABPC1L、TMEM209；PABPC1L、ARPC2。本研究结果为在不同条件下选择稻螟赤眼蜂内参基因提供了参考，也有利于在稻螟赤眼蜂的基因表达研究中取得更为可靠、准确的数据。

关键词: qRT-PCR, 稻螟赤眼蜂, 内参基因, 基因表达

Abstract: For screening suitable reference genes used in the studies of Trichogramma japonicum,ten candidate reference genes (including PABPC1L, ACTR2, ARPC2, EF1a, EEF1D, SFRS7, HSP70, CSNK1a1, GAPDH, TMEM209),selected on the base of T.japonicum transcriptome data,were evaluated for the expression stability under different conditions such as temperatures,diets,insecticides and developmental duration.The stabilities were analyzed by Delta CT,BestKeeper,NormFinder and GeNorm and finally sorted by RefFinder.The results indicated that the genes EF1a,EEF1D,SFRS7,ARPC2,PABPC1L,TMEM209,PABPC1L and ARPC2 were expressed the most stable at different temperatures,when fed with different diets,under different insecticide treatments,and at different developmental durations,respectively.The results of this study can help select reference genes in T.japonicum under different conditions,and also help obtain more reliable and accurate data for gene expression in T.japonicum.

Key words: qRT-PCR, Trichogramma japonicum, reference gene, gene expression

中图分类号:

S476.3

王珊珊, 田俊策, 鲁艳辉, 徐红星, 王香萍, 吕仲贤. 稻螟赤眼蜂实时荧光定量PCR内参基因的筛选[J]. 中国生物防治学报, 2022, 38(5): 1156-1165.

WANG Shanshan, TIAN Junce, LU Yanhui, XU Hongxing, WANG Xiangping, LÜ Zhongxian. Screening for qRT-PCR Reference Genes in Trichogramma japonicum[J]. Chinese Journal of Biological Control, 2022, 38(5): 1156-1165.

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