4.6 Article

Effects of inhibitors on the protease profiles and degradation of activated Cry toxins in larval midgut juices of Cnaphalocrocis medinalis (Lepidoptera: Pyralidae)

Journal

JOURNAL OF INTEGRATIVE AGRICULTURE
Volume 20, Issue 8, Pages 2195-2203

Publisher

ELSEVIER SCI LTD
DOI: 10.1016/S2095-3119(20)63316-0

Keywords

Cnaphalocrocis medinalis; midgut juice; protease inhibitor; enzyme activity; degradation; Bt protein

Funding

  1. Zhejiang Provincial Natural Science Foundation of China [LY20C140004]
  2. National Science and Technology Major Project of China [2016ZX08001001]
  3. National Natural Science Foundation of China [31501669]
  4. earmarked fund for China Agriculture Research System [CARS-01-36]

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Our study found that certain protease inhibitors can hinder various enzymatic activities in the larval midgut of Cnaphalocrocis medinalis, potentially reducing the insect's ability to degrade Bt toxins. These findings may help in the future application of protease inhibitors in managing this insect pest.
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 (Guenee) 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.

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