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Mol Ecol


Title:Oryzacystatin I expressed in transgenic potato induces digestive compensation in an insect natural predator via its herbivorous prey feeding on the plant
Author(s):Bouchard E; Cloutier C; Michaud D;
Address:"Departement de Biologie, Centre de recherche en horticulture, Universite Laval, Cite Universitaire (Quebec), Canada G1K 7P4"
Journal Title:Mol Ecol
Year:2003
Volume:12
Issue:9
Page Number:2439 - 2446
DOI: 10.1046/j.1365-294x.2003.01919.x
ISSN/ISBN:0962-1083 (Print) 0962-1083 (Linking)
Abstract:"We observed recently that the rice cysteine proteinase inhibitor, oryzacystatin I (OCI) expressed in transgenic potato does not affect growth and development of the two-spotted stinkbug predator (Perillus bioculatus) via its herbivorous prey feeding on the plant. Here we monitored the inhibitory activity of recombinant OCI along this potato --> herbivore --> predator continuum, to determine if the absence of effect was associated with a digestive compensatory response of the predator following inhibition of its proteinases by the recombinant cystatin. After confirming that OCI is present in the plant, and ingested in an active form by potato beetle larvae, quantitative and electrophoretic assays allowed us to determine that the recombinant cystatin (representing about 0.8% of total soluble proteins in leaves) was entirely bound to a approximately 30-kDa target proteinase in the prey's midgut, forming a sodium dodecyl sulphate (SDS)-stable complex detected on immunoblots with an anti-OCI polyclonal antibody. Despite the apparent absence of free, residual OCI in the beetle's midgut, digestive protease activity in the predator, known to include OCI-sensitive activity, was altered negatively when the prey was fed the modified plant. This inhibitory process at the third trophic level was accompanied by a compensatory response in the predator, by which serine-type proteinases were synthesized de novo. Overall, our data suggest that the affinity between OCI and the predator's OCI-sensitive proteinases is: (i) as strong as (or stronger than) the affinity between OCI and the potato beetle 30-kDa-sensitive proteinase; and (ii) stronger than the affinity between these enzymes and the plant endogenous homologue of OCI, potato multicystatin, induced in the plant by potato beetle feeding. Our results also show that predatory organisms can adapt their digestive metabolism to the presence of plant antidigestive proteins ingested by their herbivorous preys. In a broader context, this study stresses the need to monitor the inhibitory effects of PI-expressing plants not only on the herbivorous insects targeted, but also on the organisms likely to consume these pests in the environment"
Keywords:"Animals Coleoptera/*metabolism Cystatins/*metabolism Cysteine Endopeptidases/*metabolism Electrophoresis, Polyacrylamide Gel *Food Chain Hemiptera/chemistry/*metabolism Immunoblotting Plants, Genetically Modified/genetics Solanum tuberosum/*genetics/metab;"
Notes:"MedlineBouchard, Edith Cloutier, Conrad Michaud, Dominique eng Comparative Study Research Support, Non-U.S. Gov't England 2003/08/16 Mol Ecol. 2003 Sep; 12(9):2439-46. doi: 10.1046/j.1365-294x.2003.01919.x"

 
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