四川草地贪夜蛾种群对高效氯氰菊酯的抗药性及代谢途径

陈慧淋; 吴玉童; 高节; 徐翔; 王学贵; 杨继芝

doi:10.7671/j.issn.1001-411X.202501008

四川草地贪夜蛾种群对高效氯氰菊酯的抗药性及代谢途径

Resistance and metabolic pathway of Spodoptera frugiperda population to beta cypermethrin in Sichuan

摘要

摘要:
目的探究四川地区草地贪夜蛾Spodoptera frugiperda种群对高效氯氰菊酯的抗药性及代谢途径。
方法采用点滴法测定四川地区草地贪夜蛾米易(MY)、德昌(DC)、苍溪(CX)、会东(HD)和仁和(RH)种群对高效氯氰菊酯的抗药性水平，采用光电比色法和RT-qPCR测定细胞色素P450酶(Cytochrome P450 enzymes，P450s)、羧酸酯酶(Carboxylesterases，CarE)、谷胱甘肽转移酶(Glutathione S-transferase，GST) 3种解毒代谢酶的活性及基因表达量，采用皮尔逊相关性系数分析高效氯氰菊酯抗药性与酶活性、基因表达的相关性，并通过液相色谱−质谱(High performance liquid chromatography-mass spectrometry，HPLC-MS)分析经LD₅₀高效氯氰菊酯处理的MY种群(试验组)和未经处理的MY种群(对照组)的高效氯氰菊酯代谢产物差异。
结果 MY种群对高效氯氰菊酯的抗性倍数(Resistance ratio, RR)最高，为4.02倍，其LD₅₀为84.201 μg/g。CarE、GST、P450s活性均随抗性升高而升高，CES12、GST epsilon9、CYP6B50基因表达量显著上调且与高效氯氰菊酯抗药性显著相关。对照组和试验组代谢差异产物有3−苯氧基苯甲酸(3-PBA)、邻苯二酚、癸酸、甲基−2, 3−二氢−3, 5−二羟基−2−氧代−3−吲哚乙酸。其中，3-PBA只在试验组中检出，在对照组中未被检出，推断高效氯氰菊酯通过草地贪夜蛾体内酶将高效氯氰菊酯代谢为3-PBA，3-PBA在单加氧酶催化下，在苯环上引入2个羟基，生成邻苯二酚。结论草地贪夜蛾可能通过上调解毒代谢酶相关基因的表达，提高酶活性，从而增强对高效氯氰菊酯的解毒代谢能力，降低农药的毒效。

Abstract:
Objective To investigate the resistance and metabolic pathways of beta cypermethrin in fall armyworm (Spodoptera frugiperda) populations in Sichuan.
Method The resistance levels of fall armyworm to beta cypermethrin in MY, DC, CX, HD and RH populations in Sichuan were determined by drip method. The activities and gene expressions of three kinds of detoxification metabolic enzymes, such as cytochrome P450 enzymes (P450s), carboxylesterases (CarE) and glutathione S-transferase (GST), were determined by photoelectric colorimetric method and RT-qPCR. The Pearson correlation coefficient was used to analyze the correlation of beta cypermethrin resistance with enzyme activity and gene expressions. The differences in beta cypermethrin metabolic products between the MY population treated with LD₅₀ beta cypermethrin (the experimental group) and the untreated MY population (the control group) were analyzed by high performance liquid chromatography-mass spectrometry (HPLC-MS).
Result The MY population showed the highest resistance ratio (RR) to beta cypermethrin, the RR was 4.02 times and the LD₅₀ was 84.201 μg/g. The activities of CarE, GST and P450s increased with the increase of beta cypermethrin resistance, and the expression levels of CES12, GST epsilon9 and CYP6B50 genes were significantly up-regulated and significantly correlated with beta cypermethrin resistance. The metabolism differential products in the control group and the experimental group were 3-phenoxybenzoic acid (3-PBA), pyrocatechol, capric acid, methyl-2, 3-dihydro-3, 5-dihydroxy-2-oxo-3-indoleacetic acid. Among them, 3-PBA was only detected in the experimental group and not in the control group. It was inferred that beta cypermethrin was metabolized into 3-PBA through the enzyme activity of fall armyworm, 3-PBA was catalyzed by monooxygenase, two hydroxyl groups were introduced into benzene ring, and then pyrocatechol was finally generated.
Conclusion Spodoptera frugiperda may up-regulate the expression of genes associated with detoxification enzymes, leading to increased enzyme activities. This enhanced detoxification capacity accelerates the metabolic degradation of beta cypermethrin, thereby reducing the pesticide’s efficacy.

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