Analysis of rice genes induced by striped stemborer (Chilo suppressalis) attack identified a promoter fragment highly specifically responsive to insect feeding

Plant Mol Biol. 2007 Nov;65(4):519-30. doi: 10.1007/s11103-007-9185-4. Epub 2007 May 24.

Abstract

The objective of this study was to identify rice gene promoters that are specifically induced by feeding of the striped stemborer (Chilo suppressalis). Two PCR-selected cDNA subtractive libraries were constructed from the rice variety Minghui 63. Up- and down-regulated cDNAs induced by C. suppressalis feeding were arrayed on nylon membranes. After array hybridization and Northern blot analysis, a cDNA (B1-A04) encoding a putative subtilisin/chymotrypsin inhibitor was found to be rapidly and highly induced by C. suppressalis feeding, compared with mechanical wounding. The putative promoter region, spanning from -1,569 to +446 relative to the transcriptional initiation site was isolated, fused to the GUS gene (beta-glucuronidase reporter gene) and introduced by Agrobacterium-mediated transformation to rice. In non-infested plants, the GUS activity driven by this promoter fragment was detected in culms and panicles, but not in leaves and sheaths. At 6 h after insect feeding, GUS activity was significantly induced in sheaths and culms, but not in leaves. GUS activity and native B1-A04 gene were not induced by JA and ABA treatment. A serial deletion analysis revealed two regions (-1,569 to -1,166 and -1,166 to -582) that negatively regulate the gene expression in sheaths of non-infested plants but not in insect-infested plants. An electrophoretic mobility shift assay (EMSA) identified 7 DNA fragments with various binding activities with nuclear proteins from mechanically wounded, insect-infested and untreated plants, and their possible roles in gene regulation were speculated. This promoter fragment should have utility in development of insect resistant transgenic crops.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Blotting, Northern
  • Blotting, Southern
  • DNA Primers
  • DNA, Complementary
  • Feeding Behavior*
  • Genes, Plant*
  • Insecta / physiology*
  • Molecular Sequence Data
  • Oryza / genetics*
  • Oryza / parasitology
  • Promoter Regions, Genetic*
  • Transformation, Genetic

Substances

  • DNA Primers
  • DNA, Complementary