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Negative cross-resistance of a pyrethroid-resistant Drosophila mutant to Phryma leptostachya-derived haedoxan A

Insect Science 2022年第3期29卷 817-826页

作者： Xingtao Qie Yuzhe Du ahmed a.a.aioub Ke Dong Zhaonong Hu Institution of Pesticide Science College of Plant ProtectionNorthwest A&F UniversityYanglingShaanxi 712100China Key Laboratory of Botanical Pesticide R&D in Shaanxi Province YanglingShaanxi 712100China Southern Insect Management Research Unit Agriculture Research ServiceUnited States Department of Agriculture141 Experiment Station RoadStonevilleMS 38776USA Plant Protection Department Faculty of AgricultureZagazig UniversityZagazig44511 Egypt Department of Biology Duke UniversityDurhamNC 27708USA

Voltage-gated sodium channels are the primary target of pyrethroid insecticides.Mutations in sodium channel confer knockdown resistance(kdr)to pyrethroids in various arthropod pests.Haedoxan A(HA)is the major insecticidal component from Phryma leptostachya.It has been shown that HA alters electrical responses at the Drosophila neuromuscular junction and modifies the gating properties of cockroach sodium channels expressed in Xenopus oocytes.However,whether sodium channel mutations that confer pyrethroid resistance also affect the action of HA is unknown.In this study,we conducted bioassays using HA and permethrin in two Drosophila melanogaster strains:w^(1118),an insecticide-susceptible strain,and para^(tsl),a pyrethroid-resistant strain due to a I265N mutation in the sodium channel,and identified a new case of negative cross-resistance(NCR)between permethrin and HA.Both para^(tsl) larvae and adults were more resistant to permethrin,as expected.However,both para^(tsl) larvae and adults were more sensitive to HA compared to w^(1118).We confirmed that the I265N mutation reduced the sensitivity to permethrin of a Drosophila sodium channel variant,DmNa_(v)22,expressed in Xenopus oocytes.Interestingly,the I265N mutation also abolished the effect of HA on sodium channels.Further characterization showed that I265 on the sodium channels is critical for the action of both pyrethroids and HA on sodium channels,pointing to an overlapping mode of action between pyrethroids and HA on the sodium channel.Overall,our results suggest an I265N-independnt mechanism(s)in para^(tsl) flies that is responsible for the NCR between permethrin and HA at the whole insect level.

关键词： botanical insecticide haedoxan A insecticidal mechanism negative crossresistance permethrin voltage-gated sodium channel

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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 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

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