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Bacillus thuringiensis CryIA Protein Levels in Raw and Processed Seed of Transgenic Cotton: Determination Using Insect Bioassay and ELISA

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Abstract:

The levels of the Bacillus thuringiensis Berliner variety kurstaki CryIA(b) and CryIA(c) insecticidal proteins in unprocessed (raw) and processed seed of transgenic cotton were evaluated using insect bioassay and enzyme-linked immunosorbent assay (ELISA) procedures. Larvae of the tobacco budworm, He/iothis virescens (F.), were exposed to sublethal concentrations of ground cottonseed or purified CryIA proteins incorporated into artificial insect diet. The dose-response function of treatments was fit using nonlinear regression analysis to calculate EC50 (dose reducing larval growth by 50%) values and 95% CIs. The bioassay detection limit for both CryIA(b) and CryIA(c) proteins was 0.5 ng/ml insect diet. At 0.5 ng/ml, H. virescens larval weight was consistently reduced by ≥9% compared with the associated control weight. ELISA analyses of CryIA(b) and CryIA(c) protein in cotton seed provided data similar to bioassay results. Cotton line 81 [CrylA(b)] contained 11.28 µg/g (bioassay) and 14.00 µg/g (ELISA) active protein in raw seed. Line 531 [CryIA(c)] contained 2.89 µg/g (bioassay) and 2.22 µg/g(ELISA) active protein in raw seed. Processing eliminated ≥97% of the active CrylA(b) and CryIA(c) proteins in the transgenic cottonseed.

Document Type: Research Article

Publication date: February 1, 1996

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  • Journal of Economic Entomology is published bimonthly in February, April, June, August, October, and December. The journal publishes articles on the economic significance of insects and is divided into the following sections: apiculture & social insects; arthropods in relation to plant disease; forum; insecticide resistance and resistance management; ecotoxicology; biological and microbial control; ecology and behavior; sampling and biostatistics; household and structural insects; medical entomology; molecular entomology; veterinary entomology; forest entomology; horticultural entomology; field and forage crops, and small grains; stored-product; commodity treatment and quarantine entomology; and plant resistance. In addition to research papers, Journal of Economic Entomology publishes Letters to the Editor, interpretive articles in a Forum section, Short Communications, Rapid Communications, and Book Reviews.
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