A survey on the off-target effects of insecticidal double-stranded RNA targeting the Hvβ'COPI gene in the crop pest Henosepilachna vigintioctopunctata through RNA-seq  

作　　者：Lü Jing Satyabrata NANDA CHEN Shi-min MEI Yang HE Kang QIU Bao-li ZHANG You-jun LI Fei PAN Hui-peng 

机构地区：[1]Key Laboratory of Bio-pesticide Innovation and Application of Guangdong Province,South China Agricultural University,Guangzhou 510642,P.R.China [2]MS Swaminathan School of Agriculture,Centurion University of Technology and Management,Paralakhemundi 761211,India [3]Institute of Insect Sciences,College of Agriculture and Biotechnology,Zhejiang University,Hangzhou 310058,P.R.China [4]Department of Plant Protection,Institute of Vegetables and Flowers,Chinese Academy of Agricultural Sciences,Beijing 100081,P.R.China

出　　处：《Journal of Integrative Agriculture》2022年第9期2665-2674,共10页农业科学学报（英文版）

基　　金：supported by the National Natural Science Foundation of China(31972269 and 32172500);the National Key R&D Program of China(2017YFD0200900)。

摘　　要：The specificity of the double-stranded RNA(dsRNA) used in the RNA interference(RNAi) technique is crucial for the success of sequence-specific gene silencing. Currently, RNAi-mediated insect control is a trending research topic.However, the off-target effects of the dsRNA in RNAi are a major concern. In this study, the ds Hvβ’COPI(coat protein complex I, β’ subunit)-treated and untreated transcriptomes of the 28-spotted potato lady beetle(Henosepilachna vigintioctopunctata) were compared to understand its off-target gene silencing effects. The RNA-seq results revealed that 63 and 44 differentially expressed genes(DEGs) were upregulated and downregulated, respectively, in the ds Hvβ’COPI treated group as compared with the control. Validation of the differential expressions of some selected DEGs via reverse transcription-quantitative PCR(RT-qPCR) analysis confirmed the reliability of the transcriptome analysis results. Further downstream analysis revealed that there were no genes homologous with Hvβ’COPI in H. vigintioctopunctata. Additionally,no genes with a >11 bp continuous match with ds Hvβ’COPI were found in the H. vigintioctopunctata transcriptome. Six genes(Hvcitron, Hvhelicase, Hvtransposase, Hvserine, Hvdynein, and Hv E3 ubiquitin) were selected to examine the offtarget activity of ds Hvβ’COPI based on their potential involvement in various H. vigintioctopunctata metabolic pathways.The severity of silencing these six off-target genes was evaluated by employing RNAi. The RNAi results confirmed the downregulation of the expression of all six genes, although there was no significant lethality. The findings of this study will be helpful in the risk analysis of future RNAi-mediated pest control experiments.

关 键 词：RNAI DSRNA off-target effects RNA-SEQ transcriptome analysis 

分 类 号：S433[农业科学—农业昆虫与害虫防治]