Expression of Cry1Ac in transgenic tobacco plants under the control of a wound-inducible promoter (AoPR1) isolated from Asparagus officinalis to control Heliothis virescens and Manduca sexta

Mol Biotechnol. 2009 Jul;42(3):341-9. doi: 10.1007/s12033-009-9168-6. Epub 2009 Apr 8.

Abstract

Expression of cry1Ac gene from Bacillus thuringiensis (Bt) was evaluated under the control of a wound-inducible AoPR1 promoter from Asparagus officinalis in transgenic tobacco plants. The leaves of transgenic plants were mechanically wounded to evaluate the activity of the AoPR1 promoter in driving the expression of Cry1Ac protein at the wound site. Our results indicate that mechanical wounding of transgenic plants was effective in inducing the expression of Cry1Ac protein. As a result of this induction, the accumulated levels of Cry1Ac protein increased during 6-72 h post-wounding period. The leaves of transgenic tobacco plants were evaluated for resistance against Heliothis virescens and Manduca sexta in insect bioassays in two different ways. The detached tobacco leaves were either fed directly to the insect larvae or they were first mechanically wounded followed by a 72 h post-wounding feeding period. Complete protection of mechanically wounded leaves of transgenic plants was observed within 24 h of the bioassay. The leaves of transgenic plants fed directly (without pre-wounding) to the larvae achieved the same level of protection between 24 and 72 h of the bioassay.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacillus thuringiensis Toxins
  • Bacterial Proteins / biosynthesis*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Blotting, Western
  • Endotoxins / biosynthesis*
  • Endotoxins / genetics
  • Endotoxins / metabolism
  • Gene Expression Regulation, Plant
  • Hemolysin Proteins / biosynthesis*
  • Hemolysin Proteins / genetics
  • Hemolysin Proteins / metabolism
  • Larva / drug effects
  • Lepidoptera / drug effects
  • Lepidoptera / pathogenicity
  • Nicotiana / genetics*
  • Nicotiana / metabolism
  • Nicotiana / parasitology
  • Pest Control, Biological / methods
  • Plant Diseases / parasitology
  • Plant Proteins / genetics*
  • Plants, Genetically Modified / genetics*
  • Plants, Genetically Modified / metabolism
  • Plants, Genetically Modified / parasitology
  • Polymerase Chain Reaction
  • Promoter Regions, Genetic

Substances

  • Bacillus thuringiensis Toxins
  • Bacterial Proteins
  • Endotoxins
  • Hemolysin Proteins
  • Plant Proteins
  • insecticidal crystal protein, Bacillus Thuringiensis
  • PR1 protein, Asparagus officinalis