title: Functional proteomics-aided selection of protease inhibitors for herbivore insect control.
abstract: Studies have reported the potential of protease inhibitors to engineer insect resistance in transgenic plants but the general usefulness of this approach in crop protection still remains to be established. Insects have evolved strategies to cope with dietary protease inhibitors, such as the use of proteases recalcitrant to inhibition, that often make the selection of effective inhibitors very challenging. Here, we used a functional proteomics approach for the 'capture' of Cys protease targets in crude protein extracts as a tool to identify promising cystatins for plant improvement. Two cystatins found to differ in their efficiency to capture Cys proteases of the coleopteran pest Leptinotarsa decemlineata also differed in their usefulness to produce transgenic potato lines resistant to this insect. Plants expressing the most potent cystatin at high level had a strong repressing effect on larval growth and leaf intake, while plants expressing the weakest cystatin showed no effect on both two parameters compared to untransformed parental line used for genetic transformation. Our data underline the relevance of considering the whole range of possible protease targets when selecting an inhibitor for plant pest control. They also confirm the feasibility of developing cystatin-expressing transgenics resistant to a major pest of potato.
authors.affiliation: Département de phytologie, Université Laval, Québec City, QC, Canada.
authors.forename: Asieh
authors.initials: A
authors.lastname: Rasoolizadeh
authors.affiliation: Département de phytologie, Université Laval, Québec City, QC, Canada.
authors.forename: Aurélie
authors.initials: A
authors.lastname: Munger
authors.affiliation: Département de phytologie, Université Laval, Québec City, QC, Canada.
authors.forename: Marie-Claire
authors.initials: MC
authors.lastname: Goulet
authors.affiliation: Département de phytologie, Université Laval, Québec City, QC, Canada.
authors.forename: Frank
authors.initials: F
authors.lastname: Sainsbury
authors.affiliation: Département de biologie, Université Laval, Québec City QC, Canada.
authors.forename: Conrad
authors.initials: C
authors.lastname: Cloutier
authors.affiliation: Département de phytologie, Université Laval, Québec City, QC, Canada.
authors.forename: Dominique
authors.initials: D
authors.lastname: Michaud
mesh_terms: D000818:Animals; D001517:Coleoptera; D015891:Cystatins / Q000302:isolation & purification*; D015853:Cysteine Proteinase Inhibitors / Q000302:isolation & purification*; D007300:Insect Control; D007306:Insecticides; D017434:Protein Structure, Tertiary; D040901:Proteomics; D011198:Solanum tuberosum / Q000235:genetics
chemical_list: D015891:Cystatins; D015853:Cysteine Proteinase Inhibitors; D007306:Insecticides