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Journal of Integrative Agriculture  2021, Vol. 20 Issue (8): 2195-2203    DOI: 10.1016/S2095-3119(20)63316-0
Special Issue: 昆虫合辑Plant Protection—Entomolgy 害虫抗药性和毒理学合辑Pest Toxicology
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Effects of inhibitors on the protease profiles and degradation of activated Cry toxins in larval midgut juices of Cnaphalocrocis medinalis (Lepidoptera: Pyralidae)
YANG Ya-jun, XU Hong-xing, WU Zhi-hong, LU Zhong-xian
State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products, Institute of Plant Protection and Microbiology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, P.R.China
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昆虫中肠液在食物消化与解毒过程中发挥重要作用,为了明确稻纵卷叶螟中肠液对Bt蛋白潜在降解能力,本文利用蛋白酶抑制剂处理研究了稻纵卷叶螟中肠液酶活特性和中肠液对Bt蛋白(Cry2A,Cry1C,Cry1Aa和Cry1Ac)的降解能力。PMSF、TLCK、Pepstatin A和Leupeptin显著抑制稻纵卷叶螟中肠总蛋白酶活性;PMSF对胰凝乳蛋白酶活性有较强的抑制作用;EDTA、PMSF、TPCK、TLCK和E-64对胰蛋白酶活性有较强的抑制作用。EDTA对稻纵卷叶螟降解Cry2A的能力具有明显的抑制作用;稻纵卷叶螟对Cry1C和Cry1Aa的降解能力不同程度地受EDTA、PMSF、TPCK、TLCK抑制;稻纵卷叶螟对Cry1Ac降解能力不同程度地受EDTA、PMSF、TPCK、TLCK、E64抑制。研究结果表明,一些蛋白酶抑制剂可以抑制稻纵卷叶螟中肠蛋白酶活性,降低昆虫降解Bt蛋白的能力。本文研究结果有助于今后开发基于蛋白酶抑制剂的害虫防控新技术。

Midgut juice plays an important role in food digestion and detoxification in insects.  In order to understand the potential of midgut juice of Cnaphalocrocis medinalis (Guenée) to degrade Bt proteins, the enzymatic activity of midgut juice and its degradation of Bt proteins (Cry2A, Cry1C, Cry1Aa, and Cry1Ac) were evaluated in this study through protease inhibitor treatments.  The activities of total protease in midgut juices were significantly inhibited by phenylmethylsulfonyl fluoride (PMSF), tosyl-L-lysine chloromethyl ketone (TLCK), pepstatin A and leupeptin.  The enzymatic activity of chymotrypsin was significantly inhibited by PMSF, and enzymatic activity of trypsin was significantly inhibited by ethylenediaminetetraacetic acid (EDTA), PMSF, tosyl phenylalanine chloromethyl ketone (TPCK), TLCK and trans-epoxysuccinyl-L-leucylamido-(4-guanidino) butane (E-64).  EDTA could significantly inhibit the degradation of Cry2A by C. medinalis.  EDTA, PMSF, TPCK, and TLCK could inhibit the degradation of Cry1C and Cry1Aa.  EDTA, PMSF, TPCK, TLCK, and E-64 could inhibit the degradation of Cry1Ac.  Our results indicated that some protease inhibitors hindered various enzymatic activities in the larval midgut of C. medinalis, which may reduce the insect’s ability to degrade Bt toxins.  These findings may aid the application of protease inhibitors in the management of this insect pest in the future.
Keywords:  Cnaphalocrocis medinalis        midgut juice        protease inhibitor        enzyme activity        degradation        Bt protein  
Received: 02 April 2020   Accepted:
Fund: This study was supported by the Zhejiang Provincial Natural Science Foundation of China (LY20C140004), the National Science and Technology Major Project of China (2016ZX08001001), the National Natural Science Foundation of China (31501669) and the earmarked fund for China Agriculture Research System (CARS-01-36).
Corresponding Authors:  Correspondence LU Zhong-xian, E-mail:   
About author:  YANG Ya-jun, E-mail:;

Cite this article: 

YANG Ya-jun, XU Hong-xing, WU Zhi-hong, LU Zhong-xian. 2021. Effects of inhibitors on the protease profiles and degradation of activated Cry toxins in larval midgut juices of Cnaphalocrocis medinalis (Lepidoptera: Pyralidae). Journal of Integrative Agriculture, 20(8): 2195-2203.

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