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Knockin of the g275e mutation of the nicotinic acetylcholine receptor (nAChR) α6 confers high levels of resistance to spinosyns in Spodoptera exigua

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Insect Science 2022年第2期29卷 478-486页

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 Institute of Pesticide Science College of Plant ProtectionNorthwest A&F UniversityYanglingShaanxi712100 China College of Plant Protection Nanjing Agricultural UniversityNanjing210095 China Key Laboratory of Crop Pest Integrated Pest Management on the Loess Plateau of Ministry of Agriculture College of Plant ProtectionNorthwest A&F UniversityYanglingShaanxi712100 China Key Laboratory for Botanical Pesticide R&D of Shaanxi Province YanglingShaanxi712100 China Plant Protection Department Faculty of AgricultureZagazig UniversityZagazig44511 Egypt

Spinosyns,including spinosad and spinetoram,act on the insect central nervous system,gradually paralyzing or destroying the target *** resistance is associated with loss-of-function mutations in the nicotinic acetylcholine receptor(nAChR)α6 subunit in a number of agricultural *** gene editing,nAChRα6 has been verified as a target for spinosyns in five insect ***,a point mutation(g275e)in exon 9 of nAChRα6 was identified in spinosad-resistant strains of Thrips palmi and Tuta *** date,no in vivo functional evidence has been obtained to support that this mutation is involved in spinosyn resistance in lepidopteran *** 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 *** transcriptase-polymerase chain reaction and sequencing confirmed that this mutation was present in exon 9 of the nAChR transcripts in the edited 275e *** 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 *** analysis showed that the resistance trait is autosomal and incompletely *** study employs a reverse genetics approach to validate the functional role played by the g275e mutation in nAChRα6 of *** 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

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