Adaptation of Spodoptera exigua (Lepidoptera: Noctuidae) to barley trypsin inhibitor BTI-CMe expressed in transgenic tobacco |
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Authors: | Lara Pilar Ortego Felix Gonzalez-Hidalgo Elena Castañera Pedro Carbonero Pilar Diaz Isabel |
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Institution: | (1) Laboratorio de Bioquímica y Biología Molecular, Dpto. de Biotecnología-UPM, E.T.S. Ingenieros Agrónomos, 28040 Madrid, Spain;(2) Dpto. de Biología de Plantas, Centro de Investigaciones Biológicas, CSIC, Velázquez 144, 28006 Madrid, Spain;(3) Centro Nacional de Biotecnología, CSIC, Campus de Cantoblanco, 28049 Madrid, Spain |
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Abstract: | Nicotiana tabacum plants were transformed with the cDNA of barley trypsin inhibitor BTI-CMe under the control of the 35S CaMV promoter. Although the transgene was expressed and the protein was active in the homozygous lines selected, growth of Spodoptera exigua (Lepidoptera: Noctuidae) larvae reared on transgenic plants was not affected. The protease activity in larval midgut extracts after 2 days feeding on transformed tobacco leaves from the highest expressing plant showed a reduction of 25% in the trypsin-like activity compared to that from insects fed on non-transformed controls. The susceptibility of digestive serine-proteases to inhibition by BTI-CMe was confirmed by activity staining gels. This decrease was compensated with a significant induction of leucine aminopeptidase-like and carboxipeptidase A-like activities, whilechymotrypsin-, elastase-, and carboxipeptidase B-like proteases were not affected. |
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Keywords: | adaptation barley trypsin inhibitor bioassay digestive protease activity Spodoptera exigua transgenic tobacco |
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