稻螟赤眼蜂实时荧光定量PCR内参基因的筛选  

Screening for qRT-PCR Reference Genes in Trichogramma japonicum

作　　者：王珊珊 田俊策 鲁艳辉[2] 徐红星[1] 王香萍 吕仲贤[2] WANG Shanshan;TIAN Junce;LU Yanhui;XU Hongxing;WANG Xiangping;LÜZhongxian(Forewarning and Management of Agricultural and Forestry Pests,Hubei Engineering Technology Center,Yangtze University,Jingzhou 434025,China;State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products/Institute of Plant Protection and Microbiology,Zhejiang Academy of Agricultural Sciences,Hangzhou 310021,China)

机构地区：[1]农林病虫害预警与调控湖北省工程中心,长江大学,荆州434025 [2]浙江省农业科学院植物保护与微生物研究所/农产品质量安全危害因子与风险防控国家重点实验室,杭州310021

出　　处：《中国生物防治学报》2022年第5期1156-1165,共10页Chinese Journal of Biological Control

基　　金：浙江省重点研发计划(2020C02001);现代农业产业技术体系(CARS-01)。

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、Ge Norm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。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.

关 键 词：QRT-PCR 稻螟赤眼蜂 内参基因 基因表达 

分 类 号：S476.3[农业科学—农业昆虫与害虫防治]