Selection and validation of candidate reference genes for gene expression analysis in Callosobruchus chinensis(Coleoptera:Bruchidae)by qRT-PCR
Callosobruchus chinensis is a worldwide storage pest,which has a serious impact on the yield and quality of legume crops such as mung bean and adzuki bean.This paper aims to selection and validation of endogenous reference genes for analyzing the stable expression of C.chinensis in different development stages and tissues of male and female adults and different sex tissues.According to literature reports and the results of C.chinensis transcriptome sequencing,eight candidate endogenous reference genes(β-Actin,β-Tubulin,α-Tubulin,AK,GAPDH,RPL40,Hsc70,eEF1-α)were screened,and the expression of each candidate gene in C.chinensis different developmental stages(larvae,pupae,female and male adults)and adults different tissues(antennae,head,abdomen,foot,and wings)was analyzed by qRT-PCR.These candidate genes expression stabilities were evaluated by using four software-based(△Ct,geNorm,NormFinder and BestKeeper)and one web-based(RefFinder)comprehensive analysis tools.The results indicated that the Ct values of the 8 candidate endogenous reference genes were between 15～30 in different stages of development of C.chinensis and different tissues of adult.Through the comprehensive analysis of the stability of the endogenous reference gene by various software,the endogenous reference genes in different stages development and different tissues of adults in C.chinensis are recommended to use β-Act,β-Tub and β-Tub,α-Tub.The results of this study are conducive to obtaining more reliable and accurate data in the expression of C.chinensis gene of interest.
万方数据
维普
