Stability evaluation of reference genes for real-time quantitative PCR normalization in Spodoptera frugiperda(Lepidoptera:Noctuidae)  

作　　者：SHU Ben-shui YU Hai-kuo DAI Jing-hua XIE Zi-ge QIAN Wan-qiang LIN Jin-tian 

机构地区：[1]Guangzhou City Key Laboratory of Subtropical Fruit Trees Outbreak Control,Zhongkai University of Agriculture and Engineering,Guangzhou 510225,P.R.China [2]Agricultural Genomics Institute at Shenzhen,Chinese Academy of Agricultural Sciences,Shenzhen 518120,P.R.China

出　　处：《Journal of Integrative Agriculture》2021年第9期2471-2482,共12页农业科学学报（英文版）

基　　金：financially supported by the fund from the KeyArea Research and Development Program of Guangdong Province,China(2020B020223004);the Innovation Team Project in Guangdong Provincial Department of Education(2017KCXTD018);the Guangzhou Science and Technology Plan Projects,China(201704020190,201805010008 and 201904010135)。

摘　　要：Real-time quantitative PCR(qPCR)is a reliable and widely used technique for analyzing the expression profiles of target genes in different species,and reference genes with stable expressions have been introduced for the normalization of the data.Therefore,stability evaluation should be considered as the initial step for qPCR experiments.The fall armyworm Spodoptera frugiperda(J.E.Smith)(Lepidoptera:Noctuidae)is a polyphagous pest that consumes many plant species and seriously threatens corn production around the world.However,no studies thus far have examined the stability of reference genes in this pest.In this study,the expression profiles of the eight candidate reference genes of Actin,elongation factor 1 alpha(EF1α),elongation factor 2(EF2),glyceraldehyde-3-phosphate dehydrogenase(GAPDH),ribosomal protein L3(RPL3),ribosomal protein L13(RPL13),alpha-tubulin(α-TUB),and beta-1-tubulin(β-1-TUB)were obtained from S.frugiperda in different samples and the stability was evaluated byΔCt,BestKeeper,geNorm,NormFinder,and RefFinder methods.The results of pairwise variation(V)calculated by GeNorm indicated two reference genes could be selected for normalization.Therefore,the combinations of the most stable reference genes for different experimental conditions of S.frugiperda were shown as follows:EF2 and RPL13 for developmental stages,RPL3 andβ-1-TUB for larval tissue samples,EF2 and EF1αfor the larval samples treated with different temperatures,RPL3 and EF1αfor the larval samples under starvation stress,and RPL13 and EF1αfor all the samples.Our results lay the foundation for the normalization of qPCR analyses in S.frugiperda and could help guarantee the accuracy of subsequent research.

关 键 词：Spodoptera frugiperda reference genes qPCR stability evaluation different experimental conditions 

分 类 号：S433.4[农业科学—农业昆虫与害虫防治]