Rational molecular design and genetic engineering of herbicide resistant crops by structure modeling and site-directed mutagenesis of acetohydroxyacid synthase

J Mol Biol. 1996 Oct 25;263(2):359-68. doi: 10.1006/jmbi.1996.0580.

Abstract

Plants with specific resistance to a single class of herbicides have been genetically engineered by introduction of genes encoding rationally designed mutant acetohydroxyacid synthase (AHAS) enzymes. Suitable substitution mutations were identified from a three-dimensional model of an AHAS-inhibitor complex. The structural model was generated based on homology to pyruvate oxidase and an imidazolinone inhibitor was positioned in the proposed binding site using structure-activity data for this class of herbicide. Biochemical analysis of the mutant proteins expressed in Escherichia coli enabled iterative optimization of the mutant genes. Expression of recombinant proteins in tobacco plants conferred resistance in vivo. The novel approach coupling molecular modeling and molecular biology has many advantages over traditional random mutagenesis and selection methods and will be crucial to the future development for environmentally safe and sustainable agricultural systems.

MeSH terms

  • Acetolactate Synthase / genetics*
  • Amino Acid Sequence
  • Drug Resistance / genetics*
  • Genetic Engineering
  • Herbicides / toxicity*
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Plants, Genetically Modified / enzymology
  • Plants, Genetically Modified / genetics*

Substances

  • Herbicides
  • Acetolactate Synthase