Knockin of the G275E mutation of the nicotinic acetylcholine receptor (nAChR) α6 confers high levels of resistance to spinosyns in Spodoptera exigua  

作　　者：Ya-Yun Zuo Yu-Xin Xue Ze-Yu Wang Xuan Ren Ahmed A.A.Aioub Yi-Dong Wu Yi-Hua Yang Zhao-Nong Hu 

机构地区：[1]Institute of Pesticide Science,College of Plant Protection,Northwest A&F University,Yangling,Shaanxi,712100 China [2]College of Plant Protection,Nanjing Agricultural University,Nanjing,210095 China [3]Key Laboratory of Crop Pest Integrated Pest Management on the Loess Plateau of Ministry of Agriculture,College of Plant Protection,Northwest A&F University,Yangling,Shaanxi,712100 China [4]Key Laboratory for Botanical Pesticide R&D of Shaanxi Province,Yangling,Shaanxi,712100 China [5]Plant Protection Department,Faculty of Agriculture,Zagazig University,Zagazig,44511 Egypt

出　　处：《Insect Science》2022年第2期478-486,共9页昆虫科学（英文版）

基　　金：funded by the National Natural Science Foundation of China(32001941 and 31972303);the China Postdoctoral Science Foundation(2020M683586);the Research Fund for the Doctoral Program of Northwest A&F University(Z1090219195).

摘　　要：Spinosyns,including spinosad and spinetoram,act on the insect central nervous system,gradually paralyzing or destroying the target insect.Spinosad resistance is associated with loss-of-function mutations in the nicotinic acetylcholine receptor(nAChR)α6 subunit in a number of agricultural pests.Using gene editing,nAChRα6 has been verified as a target for spinosyns in five insect species.Recently,a point mutation(G275E)in exon 9 of nAChRα6 was identified in spinosad-resistant strains of Thrips palmi and Tuta absoluta.To date,no in vivo functional evidence has been obtained to support that this mutation is involved in spinosyn resistance in lepidopteran pests.In this study,the G275E mutation was introduced into the nAChR of Spodoptera exigua using clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated protein 9(Cas9)gene-editing technology.Reverse transcriptase-polymerase chain reaction and sequencing confirmed that this mutation was present in exon 9 of the nAChR transcripts in the edited 275E strain.The results of bioassays showed that the 275E strain was highly resistant to spinosad(230-fold)and spinetoram(792-fold)compared to the unedited background strain,directly confirming that the G275E mutation of the nAChRα6 subunit confers high levels of spinosyn resistance in S.exigua.Inheritance analysis showed that the resistance trait is autosomal and incompletely recessive.This study employs a reverse genetics approach to validate the functional role played by the G275E mutation in nAChRα6 of S.exigua in spinosyns resistance and provides another example of the use of CRISPR/Cas9 gene-editing technology to confirm the role played by candidate target site mutations in insecticide resistance.

关 键 词：beet armyworm CRISPR G275E mutation nicotinic acetylcholine recep・tor spinosyns insecticides target resistance 

分 类 号：S48[农业科学—农药学]