三种杀虫剂亚致死剂量对草地贪夜蛾细胞色素P450基因表达的影响

doi:10.16380/j.kcxb.2020.05.005

昆虫学报 ›› 2020, Vol. 63 ›› Issue (5): 565-573.doi: 10.16380/j.kcxb.2020.05.005

三种杀虫剂亚致死剂量对草地贪夜蛾细胞色素P450基因表达的影响

张百重^1,#, 苏栩^1,#, 卢留洋¹, 甄丛爱², 朱斌², 李亚设¹, 董文阳², 汪耿¹, 胥燕博¹, 孔凡彬¹, 刘润强¹, 陈锡岭^1,*, 高希武^2,*

(1. 河南科技学院资源与环境学院, 河南新乡 453003; 2. 中国农业大学昆虫学系, 北京 100193)

出版日期:2020-05-20 发布日期:2020-06-08

Effects of three insecticides at the sublethal dose on the expression of cytochrome P450 genes in Spodoptera frugiperda (Lepidoptera: Noctuidae)

ZHANG Bai-Zhong^1,#, SU Xu^1,#, LU Liu-Yang¹, ZHEN Cong-Ai², ZHU Bin², LI Ya-She¹, DONG Wen-Yang², WANG Geng¹, XU Yan-Bo¹, KONG Fan-Bin¹, LIU Run-Qiang¹, CHEN Xi-Ling^1,*, GAO Xi-Wu^2,*

(1. College of Resources and Environment, Henan Institute of Science and Technology, Xinxiang, Henan 453003, China; 2. Department of Entomology, China Agricultural University, Beijing 100193, China)

Online:2020-05-20 Published:2020-06-08

摘要/Abstract

摘要： 【目的】为明确杀虫剂亚致死剂量对草地贪夜蛾Spodoptera frugiperda细胞色素P450基因表达的影响。【方法】本研究采用叶片浸渍法测定了3种杀虫剂［氯虫苯甲酰胺、甲氨基阿维菌素苯甲酸盐和苏云金杆菌Bacillus thuringiensis (Bt)］对草地贪夜蛾2龄幼虫的毒力，以及通过实时荧光定量PCR(real-time quantitative PCR, RT-qPCR)技术测定了这3种杀虫剂亚致死剂量(LC₁₀)处理后48 h时草地贪夜蛾2龄幼虫16个P450基因的表达量。【结果】氯虫苯甲酰胺、甲氨基阿维菌素苯甲酸盐和Bt对草地贪夜蛾2龄幼虫的LC₁₀值分别为0.931, 0.283和1 089.688 mg/L。2龄幼虫受LC₁₀氯虫苯甲酰胺胁迫后，13个P450基因(CYP4G75, CYP6AB12, CYP6B50, CYP321A7, CY321A8, CYP321A9, CYP321A10, CYP321B1, CYP337B5, CYP9A59, CYP9A58, CYP6AE44及CYP6AE43)表达上调，其中CYP6AE44表达量为对照的34.60倍；2龄幼虫受LC₁₀甲维盐胁迫后，11个P450基因(CYP4G75, CYP6AB12, CYP321A7, CY321A8, CYP321A9, CYP321A10, CYP321B1, CYP337B5, CYP9A58, CYP6AE44及CYP6AE43)表达上调，其中CYP321B1表达量为对照的28.70倍；2龄幼虫受LC₁₀Bt胁迫后，11个P450基因(CYP4G75, CYP6AB12, CYP6AN4, CYP321A7, CY321A8, CYP321A9, CYP321A10, CYP321B1, CYP337B5, CYP6AE44及CYP6AE43)表达上调，其中CYP6AE44表达量为对照的40.80倍。【结论】草地贪夜蛾2龄幼虫的多个P450基因受这3种杀虫剂亚致死剂量处理后表达上调，其中CYP4G75, CYP6AB12, CYP321A7, CY321A8, CYP321A9, CYP321A10, CYP321B1, CYP321B5, CYP6AE44及CYP6AE43均能被这3种杀虫剂诱导表达。

关键词: 草地贪夜蛾, 细胞色素P450, 杀虫剂, 苏云金杆菌, 基因表达, 亚致死剂量

Abstract: 【Aim】 To determine the effects of insecticides at the sublethal dose on the expression of cytochrome P450 genes in the fall armyworm, Spodoptera frugiperda. 【Methods】 The toxicity of three insecticides (chlorantraniliprole, emamectin benzoate and Bacillus thuringiensis) against the 2nd instar larvae of S. frugiperda was assayed by leaf-dipping method, and the transcriptional levels of 16 P450 genes in the 2nd instar larvae of S. frugiperda treated with the sublethal dose (LC₁₀) of the three insecticides were measured by real-time quantitative PCR (RT-qPCR) at 48 h post treatment. 【Results】 The LC₁₀ values of chlorantraniliprole, emamectin benzoate and B. thuringiensis against the 2nd instar larvae of S. frugiperda were 0.931, 0.283, and 1 089.688 mg/L, respectively. In the 2nd instar larvae exposed to the LC₁₀ of chlorantraniliprole, 13 P450 genes including CYP4G75, CYP6AB12, CYP6B50, CYP321A7, CY321A8, CYP321A9, CYP321A10, CYP321B1, CYP337B5, CYP9A59, CYP9A58, CYP6AE44, and CYP6AE43 were up-regulated, and the transcription level of CYP6AE44 was 34.60-fold as high as that of the control. In the 2nd instar larvae exposed to the LC₁₀ of emamectin benzoate, 11 P450 genes including CYP4G75, CYP6AB12, CYP321A7, CY321A8, CYP321A9, CYP321A10, CYP321B1, CYP337B5, CYP9A58, CYP6AE44, and CYP6AE43 were up-regulated, and the expression level of CYP321B1 was 28.70-fold as high as that of the control. In the 2nd instar larvae exposed to the LC₁₀ of B. thuringiensis, 11 P450 genes including CYP4G75, CYP6AB12, CYP6AN4, CYP321A7, CY321A8, CYP321A9, CYP321A10, CYP321B1, CYP337B5, CYP6AE44, and CYP6AE43 were up-regulated, and the expression level of CYP6AE44 was 40.80-fold as high as that of the control. 【Conclusion】 Multiple P450 genes are up-regulated in the 2nd instar larvae of S. frugiperda in response to the sublethal dose of the three insecticides, and the expressions of CYP4G75, CYP6AB12, CYP321A7, CY321A8, CYP321A9, CYP321A10, CYP321B1, CYP321B5, CYP6AE44, and CYP6AE43 are induced by all the three insecticides.

Key words: Spodoptera frugiperda, cytochrome P450, insecticide; Bacillus thuringiensis, gene expression, sublethal dose

张百重, 苏栩, 卢留洋, 甄丛爱, 朱斌, 李亚设, 董文阳, 汪耿, 胥燕博, 孔凡彬, 刘润强, 陈锡岭, 高希武. 三种杀虫剂亚致死剂量对草地贪夜蛾细胞色素P450基因表达的影响[J]. 昆虫学报, 2020, 63(5): 565-573.

ZHANG Bai-Zhong, SU Xu, LU Liu-Yang, ZHEN Cong-Ai, ZHU Bin, LI Ya-She, DONG Wen-Yang, WANG Geng, XU Yan-Bo, KONG Fan-Bin, LIU Run-Qiang, CHEN Xi-Ling, GAO Xi-Wu . Effects of three insecticides at the sublethal dose on the expression of cytochrome P450 genes in Spodoptera frugiperda (Lepidoptera: Noctuidae)[J]. Acta Entomologica Sinica, 2020, 63(5): 565-573.

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三种杀虫剂亚致死剂量对草地贪夜蛾细胞色素P450基因表达的影响

Effects of three insecticides at the sublethal dose on the expression of cytochrome P450 genes in Spodoptera frugiperda (Lepidoptera: Noctuidae)

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