Spinosyns,including spinosad and spinetoram,act on the insect central ner-vous 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,nAChR α6 has been ver-ified as a target for spinosyns in five insect species.Recently,a point mutation(G275E)in exon 9 of nAChR α6 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 nAChR α6 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 ap-proach to validate the functional role played by the G275E mutation in nAChR α6 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.

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

Institute of Pesticide Science,College of Plant Protection,Northwest A&F University,Yangling,Shaanxi,712100,China;College of Plant Protection,Nanjing Agricultural University,Nanjing,210095,China;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;Key Laboratory for Botanical Pesticide R&D of Shaanxi Province,Yangling,Shaanxi,712100,China;Plant Protection Department,Faculty of Agriculture,Zagazig University,Zagazig,44511,Egypt

中国昆虫科学（英文版）

[1]Ya-Yun Zuo;Yu-Xin Xue;Ze-Yu Wang;Xuan Ren;Ahmed A.A.Aioub;Yi-Dong Wu;Yi-Hua Yang;Zhao-Nong Hu-.Knockin of the G275E mutation of the nicotinic acetylcholine receptor(nAChR)α6 confers high levels of resistance to spinosyns in Spodoptera exigua)[J].中国昆虫科学（英文版）,2022(02):478-486

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