中国农业科学2025，Vol.58Issue(18)：3664-3675,12.DOI:10.3864/j.issn.0578-1752.2025.18.007
沉默豆蚜细胞色素P450基因CYP6CY53和CYP302A1增强对氟啶虫酰胺的敏感性
Silencing of Cytochrome P450 Genes CYP6CY53 and CYP302A1 in Aphis craccivora Enhances the Sensitivity to Flonicamid
摘要
Abstract
[Objective]The objective of this study is to clarify the effect of key cytochrome P450(CYP450)genes in Aphis craccivora on the toxicity of flonicamid and validate their functions,thus providing a theoretical basis for managing insect resistance and formulating green control strategies that combine gene-targeted interference with pesticide treatment.[Method]P450 enzyme activity was measured after flonicamid treatment to evaluate the overall metabolic response.P450 genes of A.craccivora were identified through database mining combined with phylogenetic analysis,and the qRT-PCR was used to screen the P450 genes responsive to insecticide treatment.The protein structures of up-regulated P450 enzymes were predicted using AlphaFold 3,and molecular docking of P450 enzymes with flonicamid was conducted via AutoDock to characterize binding modes and affinities.RNA interference(RNAi)was applied to silence the target P450 genes,and then the alteration of flonicamid toxicity to A.craccivora was evaluated.[Result]After treatment with flonicamid,the activity of P450 enzymes increased significantly,peaking at 24 h,and then gradually decreased.A total of 59 P450 genes of A.craccivora were screened out,among which the expressions of CYP6CY53 in the CYP3 cluster and CYP302A1 in the Mito cluster were significantly up-regulated after flonicamid treatment,suggesting that they play a crucial role in insecticide metabolism.AlphaFold 3 predicted the structures of CYP6CY53 and CYP302A1 with confidence scores of 0.75 and 0.86,respectively.Molecular docking between the two enzymes and flonicamid was performed using AutoDock,and the results indicated that flonicamid binds to the two enzymes primarily through hydrogen bonds,with binding energy values of-15.32 and-18.17 kJ·mol-1,respectively.After silencing CYP6CY53 and CYP302A1 in A.craccivora,the median lethal concentration(LC50)of flonicamid decreased to 2.06 and 6.08 mg·L-1,respectively,and the sensitivity ratios increased to 9.26 and 3.13 times that of the control group.[Conclusion]The expression of CYP6CY53 and CYP302A1 was up-regulated in A.craccivora in response to flonicamid stress.These two genes play crucial roles in mitigating the toxicity of flonicamid,and the silencing of them can significantly enhance the sensitivity of A.craccivora to flonicamid.关键词
豆蚜/氟啶虫酰胺/P450基因/RNA干扰/毒力测定Key words
Aphis craccivora/flonicamid/P450 gene/RNA interference(RNAi)/toxicity assay引用本文复制引用
肖焯丹,乔家正,高渝岚,尚掌印,刘怀,王佳..沉默豆蚜细胞色素P450基因CYP6CY53和CYP302A1增强对氟啶虫酰胺的敏感性[J].中国农业科学,2025,58(18):3664-3675,12.基金项目
国家自然科学基金(32172393)、重庆市现代山地特色高效农业蔬菜产业技术体系项目(2020 4-6) （32172393）
