阿维菌素对瓜实蝇3种解毒酶系的影响

姜建军; 王凤英; 陈黔; 黄立飞; 曹雪梅; 杨朗

doi:10.3969/j.issn.2095-1191.2023.07.001

阿维菌素对瓜实蝇3种解毒酶系的影响

doi: 10.3969/j.issn.2095-1191.2023.07.001

姜建军^1,2,
王凤英²,
陈黔²,
黄立飞²,
曹雪梅²,
杨朗^2, ,

1 广西农业职业技术大学, 广西南宁 530007;
2 广西农业科学院植物保护研究所/农业农村部华南果蔬绿色防控重点实验室/广西作物病虫害生物学重点实验室, 广西南宁 530007

基金项目:

国家自然科学地区基金项目（31960564）；广西科技重大专项（桂科AA20108002）；广西农业科学院科技发展基金项目（桂农科2021JM72，桂农科2021YT068）

详细信息

作者简介:
姜建军(1980-),https://orcid.org/0000-0003-1836-6900,博士,副研究员,主要从事果蔬害虫抗药性监测与抗性机制研究工作,E-mail:jiangjianjun8008@126.com

通讯作者:
杨朗(1976-),https://orcid.org/0009-0009-7274-9663,博士,研究员,主要从事农业昆虫与害虫防治研究工作,E-mail:107561466@qq.com

中图分类号: S433
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出版历程
- 收稿日期: 2023-02-07

Effects of abamectin on the three detoxification enzyme systems in Bactrocera cucurbitae(Coquillett)

1 Guangxi Vocational University of Agriculture, Nanning, Guangxi 530007, China;
2 Plant Protection Research Institute, Guangxi Academy of Academy of Agricultural Science/Key Laboratory of Green Prevention and Control on Fruits and Vegetables in South China, Ministry of Agriculture and Rural Affairs/Guangxi Key Laboratory of Biology for Crop Diseases and Insect Pests, Nanning, Guangxi 530007, China

Funds:

Regional Project of National Natural Science Foundation of China(31960564)；Guangxi Science and Technology Key Project(Guike AA20108002)；Science and Technology Development Fund of Guangxi Academy of Agricultural Sciences (Guinongke 2021JM72，Guinongke 2021YT068)

摘要

摘要: 【目的】探讨阿维菌素对瓜实蝇［Bactrocera cucurbitae（Coquillett）］解毒酶系的影响，明确瓜实蝇阿维菌素抗性与解毒酶的关系，为瓜实蝇抗药性综合治理和科学用药提供理论基础。【方法】采用饲毒法测定阿维菌素对瓜实蝇成虫敏感品系（SS）和抗阿维菌素品系（RS）的亚致死浓度（LC₂₅）及致死中浓度（LC₅₀）；利用微孔板法测定SS品系和RS品系经阿维菌素LC₂₅、LC₅₀剂量处理12、24、48和72 h后其体内谷胱甘肽S-转移酶（Glutathione S-transferase，GSTs）、羧酸酯酶（Carboxylesterase，CarE）活性，采用ELISA试剂盒法测定细胞色素P450单加氧化酶（CytochromeP450 monooxygenases，P450s）含量；测定比较SS品系和RS品系GSTs、CarE米氏常数（K_m）及最大反应速率（V_max）的变化及增效剂顺丁烯二酸二乙酯（DEM）、磷酸三苯酯（TPP）和增效醚（PBO）对阿维菌素的增效作用。【结果】与SS品系相比，RS品系体内GSTs和CarE活性均显著上升（P<0.05，下同），分别为SS品系的2.92和1.46倍，P450s含量则显著下降，为SS品系的68.0%。SS品系在LC₂₅剂量的阿维菌素处理后12、48 h及LC₅₀处理后12、24、48 h和RS品系在LC₂₅剂量处理后12 h、LC₅₀处理后72 h，其体内GSTs活性均被诱导显著增强； LC₂₅剂量处理后24 h，SS品系体内CarE活性被诱导显著增强，RS品系中LC₂₅处理后72 h和LC₅₀处理后各时间段CarE活性表现为被显著抑制； LC₅₀处理后12 h，P450s含量在SS品系中表现为显著降低，而在72 h则显著升高。与SS品系相比，RS品系GSTs的K_m显著降低，V_max显著升高，CarE的K_m和V_max变化不明显；增效剂DEM、TPP和PBO在SS品系中的增效比分别为1.03、1.04和2.08，在RS品系中的增效比分别为3.45、2.52和2.26，相对增效系数分别为0.71、0.60和0.54。【结论】 GSTs和CarE解毒酶参与了瓜实蝇对阿维菌素的抗性形成，其中GSTs活性发生了量变和质变，其对底物亲和力增加和代谢速度增快，在瓜实蝇对阿维菌素抗性形成中起到主导作用。
- 瓜实蝇 /
- 阿维菌素 /
- 解毒酶 /
- 抗药性
Abstract: 【Objective】 The purpose of the study was to explore the effects of abamectin on the detoxification enzyme system of Bactrocera cucurbitae(Coquillett), to clarify the relationship between abamectin resistance and detoxification enzymesin B. cucurbitae,and to provide theoretical basis for the comprehensive management of B. cucurbitae resistance and the scientific use of medication.【Method】 The feeding toxicity method was used to determine the sublethal concentration(LC₂₅) and lethal concentration(LC₅₀) of abamectin on susceptible strain(SS) and abamectin resistant strain(RS) of adult B. cucurbitae. Microplate method was used to determine the activities of glutathione S-transferase(GSTs) and carboxylesterase(CarE) in adults of SS and RS strains and those treated with LC₂₅ and LC₅₀ doses for 12,24,48 and 72 h. Cytochrome P450 monooxygenases(P450s) content was determined by ELISA kit method. The changes in GSTs, Michaelis constant(K_m) and maximal velocity(V_max) values and synergistic effects of synergists diethyl maleate(DEM),triphenyl phosphate(TPP) and potentiometric booster ether(PBO) on abamectin were tested and analyzed in the adults of SS and RS strains.【Result】 The activities of GSTs and CarE in the RS strain were significantly increased(P<0.05,the same below) compared with those in the SS strain,which were 2.92 and 1.46 times of that in the SS strain,while the P450 content was significantly decreased,which was 68.0% of that in the SS strain.After being exposed to LC₂₅ for 12,48 h and LC 50 for 12,24,48 h in SS strain and LC₂₅ for 12 h,LC₅₀ for 72 h in RS strain of abamectin,theactivities of GSTs were induced to be significantly enhanced. CarE activity was induced to be significantly enhanced at 24 h after LC₂₅ dose treatment in the SS strain and was significantly inhibited at 72 h after LC₂₅ treatment and at all times after LC₅₀ treatment in the RS strain. The content of P450s was decreased at 12 h after LC 50 treatment and was increased at 72 h after LC₅₀ treatment. Compared with SS strain,the K_m value of GSTs in the RS strain was significantly reduced and V_max was significant increased,and the K_m and V_max of CarE were not significantly changed. The synergistic ratios of synergists DEM,TPP and PBO were 1.03,1.04 and 2.08 in the SS strain and 3.45,2.52 and 2.26 in the RS strain,with relative synergistic coefficients of 0.71,0.60 and 0.54 respectively.【Conclusion】 The GSTs and CarE detoxification enzymes are involved in the formation of resistance to abamectin in B. cucurbitae.Among them,GSTs activity has quantitative and qualitative changes, and their increased affinity for substrates and faster metabolism play a leading role in B. cucurbitae to form abamectin metabolic resistance.
- Bactrocera cucurbitae(Coquillett) /
- abamectin /
- detoxification enzyme /
- insecticide resistance

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阿维菌素对瓜实蝇3种解毒酶系的影响

doi: 10.3969/j.issn.2095-1191.2023.07.001

作者简介:
姜建军(1980-),https://orcid.org/0000-0003-1836-6900,博士,副研究员,主要从事果蔬害虫抗药性监测与抗性机制研究工作,E-mail:jiangjianjun8008@126.com

通讯作者:
杨朗(1976-),https://orcid.org/0009-0009-7274-9663,博士,研究员,主要从事农业昆虫与害虫防治研究工作,E-mail:107561466@qq.com

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Effects of abamectin on the three detoxification enzyme systems in Bactrocera cucurbitae(Coquillett)

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留言板

阿维菌素对瓜实蝇3种解毒酶系的影响

doi: 10.3969/j.issn.2095-1191.2023.07.001

作者简介: 姜建军(1980-),https://orcid.org/0000-0003-1836-6900,博士,副研究员,主要从事果蔬害虫抗药性监测与抗性机制研究工作,E-mail:jiangjianjun8008@126.com

通讯作者: 杨朗(1976-),https://orcid.org/0009-0009-7274-9663,博士,研究员,主要从事农业昆虫与害虫防治研究工作,E-mail:107561466@qq.com

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Effects of abamectin on the three detoxification enzyme systems in Bactrocera cucurbitae(Coquillett)

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出版历程

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作者简介:
姜建军(1980-),https://orcid.org/0000-0003-1836-6900,博士,副研究员,主要从事果蔬害虫抗药性监测与抗性机制研究工作,E-mail:jiangjianjun8008@126.com

通讯作者:
杨朗(1976-),https://orcid.org/0009-0009-7274-9663,博士,研究员,主要从事农业昆虫与害虫防治研究工作,E-mail:107561466@qq.com