丝氨酸蛋白酶抑制剂Serpin1对绿僵菌侵染飞蝗的影响

doi:10.16409/j.cnki.2095-039x.2020.05.012

中国生物防治学报 ›› 2020, Vol. 36 ›› Issue (5): 729-736.DOI: 10.16409/j.cnki.2095-039x.2020.05.012

丝氨酸蛋白酶抑制剂Serpin1对绿僵菌侵染飞蝗的影响

李贝贝^1,2, 田野^1,3, 杜桂林⁴, 岳方正⁵, 农向群^1,2, 刘廷辉³, 张泽华^1,2, 王广君^1,2

1. 中国农业科学院植物保护研究所/植物病虫害生物学国家重点实验室, 北京 100193;
2. 农业农村部锡林郭勒草原有害生物科学观测实验站, 锡林浩特 026000;
3. 河北农业大学植物保护学院, 保定 071001;
4. 全国畜牧总站, 北京 100125;
5. 国家林业和草原局森林和草原防治总站, 沈阳 110034

收稿日期:2019-11-28 出版日期:2020-10-08 发布日期:2020-11-20
通讯作者: 王广君,副研究员,E-mail:wangguangjun@caas.cn。
作者简介:李贝贝,硕士研究生,E-mail:bei96633@163.com
基金资助:
国家重点研发计划（2017YFD0200402）；中央级科研院所基本科研业务费专项（Y2020XC18，S2019XM20，Y2018YJ16）；现代农业产业技术体系（CARS-34-07B）

Effect of Serine Protease Inhibitor Serpin1 from Locusta Migratoria on the Infection of Metarhizium anisopliae

LI Beibei^1,2, TIAN Ye^1,3, DU Guiling⁴, YUE Fangzheng⁵, NONG Xiangqun^1,2, LIU Tinghui³, ZHANG Zehua^1,2, WANG Guangjun^1,2

1. State Key Laboratory for Biology of Plant Diseases and Insect Pests/Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China;
2. Xilingol Grassland Pest Science Observation and Experiment Station, Ministry of Agriculture and Rural Affairs, Xilinhot 026000, China;
3. College of Plant Protection, Hebei Agricultural University, Baoding 071001, China;
4. National Animal Husbandry Service, Beijing 100125, China;
5. Forest and Grassland Control Station of State Forestry and Grassland Administration, Shenyang 110034, China

Received:2019-11-28 Online:2020-10-08 Published:2020-11-20

摘要/Abstract

摘要： 丝氨酸蛋白酶抑制剂（Serpins）是昆虫体内一类非常重要的蛋白酶抑制剂，具有调节昆虫免疫反应的功能。为研究Serpin1对绿僵菌Metarhizium anisopliae侵染飞蝗Locusta migratoria的影响，从飞蝗体内克隆了serpin1基因，经原核表达获得Serpin1蛋白；以飞蝗为试虫，通过饵剂饲喂的方法，测定了Serpin1蛋白对绿僵菌菌株IMI330189毒力的影响，并检测了绿僵菌IMI330189单独处理、Serpin1单独处理、绿僵菌IMI330189与Serpin1混合处理情况下飞蝗体内保护酶POD和SOD的活力。结果表明，在Serpin1与绿僵菌IMI330189混合饲喂3 d后，飞蝗的累计死亡率开始低于绿僵菌单独处理；到第8 d，绿僵菌IMI330189单独处理飞蝗累计死亡率达到100%，而Serpin1与绿僵菌IMI330189混合处理累计死亡率为84.31%，两者之间差异显著。POD和SOD酶活力测定结果显示，绿僵菌IMI330189单独处理时飞蝗体内的POD活力在第2 d和第3 d较对照组显著升高，而在第5 d较对照组显著下降；绿僵菌IMI330189与Serpin1混合处理后飞蝗体内的POD活力在第3 d和第5 d较对照组和绿僵菌处理组显著升高，而在第1 d显著低于对照组和绿僵菌处理组。绿僵菌IMI330189单独处理及绿僵菌IMI330189与Serpin1混合处理时，飞蝗体内的SOD活力在第1 d、第3 d和第5 d较对照组显著升高，在第2 d时较对照显著降低，且绿僵菌IMI330189与Serpin1混合处理组在第2 d和第3 d时较绿僵菌IMI330189单独处理组显著升高。上述研究结果表明Serpin1能够通过调节飞蝗体内POD和SOD的活性，有效抑制绿僵菌IMI330189对飞蝗的侵染。本研究为揭示Serpin1的生理功能奠定了基础。

关键词: 飞蝗, Serpin1, 绿僵菌, 毒力测定, 酶活力

Abstract: Serpins are a particularly important class of protease inhibitors in insects and have the function of regulating insect immune responses. Effect of Serpin1on the infection of Metarhizium anisopliae was investigated. The serpin1 gene was cloned from Locust amigratoria and expressed in Escherichia coli BL21, and then Serpin1 protein was tested for its effect on virulence of M. anisopliae IMI330189 to the locust by feeding method. The activities of protective enzymes SOD and POD were measured in the locusts treated with M. anisopliae IMI330189, Serpin1, or M. anisopliae IMI330189 plus Serpin1. The results showed that combined treatment of M. anisopliae IMI330189 and Serpin1 decreased the mortality of locust as compared to treatment of M. anisopliae alone at 3rd day post treatment (DPT). The treatment of M. anisopliae alone caused 100% mortality of locust at the 8th DPT while the combined treatment of M. anisopliae and Serpin1 resulted in a mortality of 84.31%, the difference was significant. Activity of POD in the treatment of M. anisopliae alone increased and was significantly higher than that in the control at the 2nd and 3rd DPT, and then decreased on the 5th DPT and was significantly lower than that in the control. POD activity in the combined treatment of M. anisopliae IMI330189 and Serpin1 was significantly higher than the control group and the treatment of M. anisopliae IMI330189 alone at the 3rd and 5th DPT, while was significantly lower at the 1st DPT. Activities of SOD in the treatment of M. anisopliae IMI330189 alone and the combined treatment of M. anisopliae IMI330189 and Serpin1 were significantly higher than that in the control group at the 1st, 3rd and 5th DPT while significantly lower at the 2nd DPT. SOD activity in the combined treatment of M. anisopliae IMI330189 and Serpin1 was significantly higher than that in the M. anisopliae IMI330189 treatment at the 2nd and 3rd DPT. Our results confirmed that the serpin1 could effectively inhibit the fungal infection by decreasing the activity of proteases in locust and laid a foundation for further study of the physiological functions of serpins.

Key words: Locusta migratoria, serpin1, Metarhizium anisopliae, toxicity test, enzyme activity

中图分类号:

S476.1

李贝贝, 田野, 杜桂林, 岳方正, 农向群, 刘廷辉, 张泽华, 王广君. 丝氨酸蛋白酶抑制剂Serpin1对绿僵菌侵染飞蝗的影响[J]. 中国生物防治学报, 2020, 36(5): 729-736.

LI Beibei, TIAN Ye, DU Guiling, YUE Fangzheng, NONG Xiangqun, LIU Tinghui, ZHANG Zehua, WANG Guangjun. Effect of Serine Protease Inhibitor Serpin1 from Locusta Migratoria on the Infection of Metarhizium anisopliae[J]. Chinese Journal Of Biological Control, 2020, 36(5): 729-736.

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丝氨酸蛋白酶抑制剂Serpin1对绿僵菌侵染飞蝗的影响

Effect of Serine Protease Inhibitor Serpin1 from Locusta Migratoria on the Infection of Metarhizium anisopliae

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