Engineered multidomain cysteine protease inhibitors yield resistance against western flower thrips (Frankliniella occidentalis) in greenhouse trials

N.S. Outchkourov, W.J. de Kogel, G.L. Wiegers, M. Abrahamson, M.A. Jongsma

    Research output: Contribution to journalArticleAcademicpeer-review

    53 Citations (Scopus)

    Abstract

    Western flower thrips, Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae), cause very large economic damage on a variety of field and greenhouse crops. In this study, plant resistance against thrips was introduced into transgenic potato plants through the expression of novel, custom-made, multidomain protease inhibitors. Representative classes of inhibitors of cysteine and aspartic proteases [kininogen domain 3 (K), stefin A (A), cystatin C (C), potato cystatin (P) and equistatin (EIM)] were fused into reading frames consisting of four (K-A-C-P) to five (EIM-K-A-C-P) proteins, and were shown to fold into functional inhibitors in the yeast Pichia pastoris. The multidomain proteins were expressed in potato and found to be more resistant to degradation by plant proteases than the individual domains. In a time span of 14-16 days, transgenic potato plants expressing EIMKACP and KACP at a similar concentration reduced the number of larvae and adults to less than 20?f the control. Leaf damage on protected plants was minimal. Engineered multidomain cysteine protease inhibitors thus provide a novel way of controlling western flower thrips in greenhouse and field crops, and open up possibilities for novel insect resistance applications in transgenic crops.
    Original languageEnglish
    Pages (from-to)449-458
    JournalPlant Biotechnology Journal
    Volume2
    Issue number5
    DOIs
    Publication statusPublished - 2004

    Keywords

    • proteinase-inhibitors
    • pichia-pastoris
    • reactive-site
    • equistatin
    • expression
    • potato
    • purification
    • cystatin
    • domains
    • trypsin

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