斜纹夜蛾谷胱甘肽S-转移酶SlGSTS4对拟除虫菊酯和有机磷类杀虫剂代谢和抗氧化活性

doi:10.16380/j.kcxb.2024.09.001

昆虫学报 ›› 2024, Vol. 67 ›› Issue (9): 1173-1181.doi: 10.16380/j.kcxb.2024.09.001

• 研究论文 • 下一篇

斜纹夜蛾谷胱甘肽S-转移酶SlGSTS4对拟除虫菊酯和有机磷类杀虫剂代谢和抗氧化活性

刘泽政¹, 王佳音¹, 袁迎秋¹, 徐莉¹, 刘润强¹, 徐永贵^2,*, 李冬植^1,*

(1. 河南科技学院资源与环境学院, 新乡 453003; 2. 河南省科学技术交流中心, 郑州 450008)

出版日期:2024-09-20 发布日期:2024-10-22

Metabolic and antioxidant activities of the glutathione S-transferase SlGSTS4 of Spodoptera litura (Lepidoptera: Noctuidae) against the pyrethroid and organophosphate insecticides

LIU Ze-Zheng¹, WANG Jia-Yin¹, YUAN Ying-Qiu¹, XU Li¹, LIU Run-Qiang¹, XU Yong-Gui^2,*, LI Dong-Zhi^1,*

(1. School of Resources and Environment, Henan Institute of Science and Technology, Xinxiang 453003, China; 2. Henan Science and Technology Exchange Center, Zhengzhou 450008, China)

Online:2024-09-20 Published:2024-10-22

摘要/Abstract

摘要： 【目的】探究斜纹夜蛾Spodoptera litura谷胱甘肽S-转移酶SlGSTS4在拟除虫菊酯和有机磷类杀虫剂抗性中的作用。【方法】构建pET-26b(+)/SlGSTs4表达载体，将其在大肠杆菌Escherichia coli中异源表达，获得SlGSTS4重组蛋白，测定其对模式底物1-氯-2, 4-二硝基苯(1-chlom-2, 4-dinitrobenzene, CDNB)的米氏常数(K_m)和最大反应速度(V_max)；采用超高效液相色谱(ultra-performance liquid chromatography, UPLC)测定重组蛋白SlGSTS4和表达SlGSTS4的大肠杆菌对拟除虫菊酯类杀虫剂(氰戊菊酯、高效氯氰菊酯和功夫菊酯)和有机磷类杀虫剂(毒死蜱和辛硫磷)的代谢率，并通过滤纸片抑菌圈试验测定SlGSTS4在过氧化氢异丙苯胁迫下的抗氧化活性。【结果】重组蛋白SlGSTS4 对CDNB的V_max和K_m值分别为(13.8±2.3) nmol/(min·mg)和(1.66±0.1) mmol/L。UPLC结果表明，与对照相比，SlGSTS4重组蛋白仅对功夫菊酯有一定的代谢率(4.9%)，而对氰戊菊酯、高效氯氰菊酯、辛硫磷和毒死蜱无代谢活性；转化SlGSTS4的大肠杆菌对氰戊菊酯、高效氯氰菊酯、功夫菊酯、毒死蜱和辛硫磷的代谢残留量峰面积均无明显变化，并且可以显著降低抑菌圈直径。【结论】SlGSTS4对拟除虫菊酯和有机磷类杀虫剂无直接代谢活性，具有明显的抗氧化活性。本研究结果丰富了对斜纹夜蛾GSTs Sigma家族基因的功能认知，有助于阐明SlGSTS4在斜纹夜蛾对拟除虫菊酯和有机磷类杀虫剂抗性中的功能。

关键词: 斜纹夜蛾, SlGSTS4, 拟除虫菊酯类杀虫剂, 有机磷类杀虫剂, 代谢活性, 抗氧化活性

Abstract: 【Aim】To investigate the role of the glutathione S-transferase SlGSTS4 of Spodoptera litura in the resistance to the pyrethroid and organophosphate insecticides.【Methods】The expression vector pET-26b(+)/SlGSTs4 was constructed and heterologously expressed in Escherichia coli. The recombinant SlGSTS4 protein was obtained, and the Michaelis constant K_m and maximum speed V_max of SlGSTS4 against the model substrate 1-chlom-2, 4-dinitrobenzene (CDNB) were determined. The metabolic rate of the recombinant SlGSTS4 and SlGSTS4-expressed E. coli to pyrethroid insecticides (fenvalerate, beta-cypermethrin and cyhalothrin) and organophosphate insecticides (chlorpyrifos and phoxim) were determined using ultra-performance liquid chromatography (UPLC). The antioxidant activity of SlGSTS4 under the stress of cumene hydrogen peroxide was determined by inhibition zone test using filter paper. 【Results】 The V_max and K_m values of the recombinant SlGSTS4 against CDNB were (13.8±2.3) nmol/(min·mg) and (1.66±0.1) mmol/L, respectively. The UPLC result indicated that the recombinant SlGSTS4 showed certain metabolic rate (4.9%) only to cyhalothrin, while had no metabolic activity to fenvalerate, beta-cypermethrin, phoxim or chlorpyrifos compared with the control. E. coli transformed with SlGSTS4 showed no influence on the residual peak area of fenvalerate, beta-cypermethrin, cyhalothrin, chlorpyrifos and phoxim, and decreased the inhibition zone diameter significantly compared with the control.【Conclusion】 SlGSTS4 showed no direct metabolic activity to pyrethroid or organophosphate insecticides, and exhibited obvious antioxidant activity. This study result has enriched the functional understanding of the GSTs Sigma family genes in S. litura, being helpful to clarify the function of SlGSTS4 in the resistance of S. litura to pyrethroid and organophosphate insecticides.

Key words: Spodoptera litura, SlGSTS4, pyrethroid insecticides, organophosphate insecticides, metabolic activity, antioxidant activity

刘泽政, 王佳音, 袁迎秋, 徐莉, 刘润强, 徐永贵, 李冬植. 斜纹夜蛾谷胱甘肽S-转移酶SlGSTS4对拟除虫菊酯和有机磷类杀虫剂代谢和抗氧化活性[J]. 昆虫学报, 2024, 67(9): 1173-1181.

LIU Ze-Zheng, WANG Jia-Yin, YUAN Ying-Qiu, XU Li, LIU Run-Qiang, XU Yong-Gui, LI Dong-Zhi. Metabolic and antioxidant activities of the glutathione S-transferase SlGSTS4 of Spodoptera litura (Lepidoptera: Noctuidae) against the pyrethroid and organophosphate insecticides[J]. Acta Entomologica Sinica, 2024, 67(9): 1173-1181.

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斜纹夜蛾谷胱甘肽S-转移酶SlGSTS4对拟除虫菊酯和有机磷类杀虫剂代谢和抗氧化活性

Metabolic and antioxidant activities of the glutathione S-transferase SlGSTS4 of Spodoptera litura (Lepidoptera: Noctuidae) against the pyrethroid and organophosphate insecticides

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