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Genetic engineering with α-amylase inhibitors makes seeds resistant to bruchids

Published online by Cambridge University Press:  19 September 2008

Maarten J. Chrispeels*
Affiliation:
Department of Biology, University of California San Diego, La Jolla, CA 92093-0116, USA
M. Fatima Grossi de Sa
Affiliation:
Cenargen/EMBRAPA, PO Box 10.2372, Brasilia DF, 70770, Brazil
T. J. V. Higgins
Affiliation:
Division of Plant Industry, CSIRO, GPO Box 1600, Canberra, ACT 2601, Australia
*
*+1-619-534-4052[email protected]

Abstract

Seeds of the common bean, Phaseolus vulgaris, contain two inhibitors of mammalian and insect α-amylases (αAls) that show specificity towards the amylases of different insect species. Expression in pea (Pisum sativum) and azuki bean (Vigna angularis) of a chimeric gene consisting of the cDNA of bean αAl-1 and a seed-specific promoter makes the seeds of these legumes resistant to three species of Old World bruchids whose amylases are inhibited by αAl-1. This was the first successful genetic engineering of insect resistance in seeds. To understand the basis of the specificity between amylases and inhibitors we cloned a second bean inhibitor (αAl-2) with different specificity, and we cloned the cDNA of the New World bruchid, Zabrotes subfasciatus. The amylase of this bruchid is inhibited by αAl-2, but not by αAl-1. Knowledge of the amino acid sequences and of the three-dimensional structure of the pancreatic α-amylase–αAl-1 complex allows us to predict the peptide domains and amino acids of the proteins that are important for protein–protein recognition and inhibition of enzyme activity.

Type
Research Papers
Copyright
Copyright © Cambridge University Press 1998

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