不同蛋白酶抑制剂对绿僵菌侵染东亚飞蝗的影响

doi:10.16409/j.cnki.2095-039x.2016.06.012

中国生物防治学报 ›› 2016, Vol. 32 ›› Issue (6): 756-761.DOI: 10.16409/j.cnki.2095-039x.2016.06.012

不同蛋白酶抑制剂对绿僵菌侵染东亚飞蝗的影响

王正浩^1,2,3, 贾苗^2,3, 张彦丰^2,3, 杜桂林⁴, 曹广春^2,3, 农向群^2,3, 刘长仲¹, 王广君^2,3, 张泽华^2,3

1. 甘肃农业大学植物保护学院兰州 730070;
2. 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 100081;
3. 农业部锡林郭勒草原有害生物科学观测实验站, 锡林浩特 026000;
4. 全国畜牧总站, 北京 100125

收稿日期:2016-04-10 出版日期:2016-12-08 发布日期:2016-12-07
通讯作者: 刘长仲,博士,教授,E-mail:liuchzh@gsau.edu.cn;王广君,博士,副研究员,E-mail:gjwang@ippcaas.cn
作者简介:王正浩(1991-),男,硕士研究生,E-mail:gsauwzh@163.com
基金资助:
现代农业产业技术体系（CARS-35-07）；“十二·五”农村领域国家科技计划（2011AA10A204-1，2012BAD19B0110）

Effects of Different Inhibitors on Digestive Enzyme and Detoxification Enzyme in Midgut of Locusta migratoria manilensis during Metarhizium anisopliae Infection

WANG Zhenghao^1,2,3, JIA Miao^2,3, ZHANG Yanfeng^2,3, DU Guilin⁴, CAO Guangchun^2,3, NONG Xiangqun^2,3, LIU Changzhong¹, WANG Guangjun^2,3, ZHANG Zehua^2,3

1. College of Plant Protection, Gansu Agricultural University, Lanzhou 730070, China;
2. State Key Laboratory of Plant Diseases and Insect Pests/Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100081, China;
3. Scientific Observation and Experimental Station of Pests in Xilingol Rangeland, Ministry of Agriculture, Xilinhot 026000, China;
4. National Animal Husbandry Service, Beijing 100125, China

Received:2016-04-10 Online:2016-12-08 Published:2016-12-07

摘要/Abstract

摘要： 为了揭示不同蛋白酶抑制剂对绿僵菌致病力的影响，明确绿僵菌经消化道侵染东亚飞蝗的作用机制，采用分光光度法测定了经绿僵菌IPPM202及不同蛋白酶抑制剂处理后，不同时间东亚飞蝗中肠总蛋白酶、类胰蛋白酶、类胰凝乳蛋白酶、乙酰胆碱酯酶（AchE）和谷胱甘肽硫转移酶（GSTs）活力的动态变化。结果表明，绿僵菌IPPM202处理后10 d内，东亚飞蝗中肠总蛋白酶活性呈现先上升再下降再上升的趋势，类胰蛋白酶活性呈先上升再下降趋势，类胰凝乳蛋白酶活性逐渐上升；但两种解毒酶的变化均为先上升后下降。加入两种抑制剂后，总体上抑制了东亚飞蝗中肠总蛋白酶和两种解毒酶的活性，降低了菌株IPPM202的侵染力，其中，甲苯磺酰基-L-氨基联苯氯甲基酮（tosyl-L-phenylalanine chloromethyl ketone，TPCK）作用较对脒苯基甲磺酰氯（4-amidino phenyl methane sulfonyl fluoride，APMSF）更加显著，使各种酶活力降低至与对照无差异水平。本研究对于揭示绿僵菌在中肠的致病机理及寄主对真菌侵染的免疫机制有一定的指导意义。

关键词: 绿僵菌, 东亚飞蝗, 抑制剂, 消化酶, 解毒酶

Abstract: To investigate the infection mechanism of Metarhizium anisopliae in midgut of Locusta migratoria manilensis (Meyen), effects of M. anisopliae on the activities of main digestive enzymes and detoxification enzymes in locust midgut were measured. The activities of total protease, trypsin like and chymotrypsin like enzymes, acetylcholinesterase (AChE) and glutathione-S-transferases (GSTs) were assayed to L. migratoria manilensis fed with M. anisopliae, trypsin like enzyme inhibitor (APMSF) and chymotrypsin like enzyme inhibitor (TPCK). In the M. anisopliae treated group, the total activity of protease experienced increase soon after treatment, decrease and increase again in comparison with the control within 10 days. Activities of trypsin like enzymes decreased sharply followed by a great increase first and activities of chymotrypsin like enzymes increased gradually during the whole period. The two inhibitors, especially TPCK, reduced the activities of total protease and two detoxification enzymes on the whole and suppressed the infection of M. anisopliae in the midgut of L. migratoria manilensis. These results reveal the interaction relation between M. anisopliae and hosts and the immune mechanism of hosts against pathogens.

Key words: Metarhizium anisopliae, Locusta migratoria manilensis, inhibitor, digestive enzymes, detoxification enzymes

中图分类号:

S476.12

王正浩, 贾苗, 张彦丰, 杜桂林, 曹广春, 农向群, 刘长仲, 王广君, 张泽华. 不同蛋白酶抑制剂对绿僵菌侵染东亚飞蝗的影响[J]. 中国生物防治学报, 2016, 32(6): 756-761.

WANG Zhenghao, JIA Miao, ZHANG Yanfeng, DU Guilin, CAO Guangchun, NONG Xiangqun, LIU Changzhong, WANG Guangjun, ZHANG Zehua. Effects of Different Inhibitors on Digestive Enzyme and Detoxification Enzyme in Midgut of Locusta migratoria manilensis during Metarhizium anisopliae Infection[J]. journal1, 2016, 32(6): 756-761.

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不同蛋白酶抑制剂对绿僵菌侵染东亚飞蝗的影响

Effects of Different Inhibitors on Digestive Enzyme and Detoxification Enzyme in Midgut of Locusta migratoria manilensis during Metarhizium anisopliae Infection

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