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Coleopterancidal delta-endotoxin and constructing its genomic library

X Li1, R Li

  • 1Wuhan Institute of Virology, Chinese Academy of Sciences.

Chinese Journal of Biotechnology
|January 1, 1994
PubMed
Summary

Researchers investigated five new Bacillus thuringiensis (Bt) strains for coleopterancidal crystal proteins. Strain YM-03 showed the highest toxicity against Plagiodera versicolora larvae, with distinct plasmid patterns and coleoptera-specific delta-endotoxin genes identified.

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Area of Science:

  • Microbiology
  • Insect Toxicology
  • Molecular Biology

Background:

  • Bacillus thuringiensis (Bt) is a bacterium producing insecticidal crystal proteins.
  • Coleopteran pests represent a significant agricultural challenge.
  • Understanding Bt strain diversity is crucial for developing targeted biocontrol agents.

Purpose of the Study:

  • To characterize five novel coleopterancidal Bacillus thuringiensis strains.
  • To analyze crystal protein composition and plasmid profiles.
  • To identify and clone coleoptera-specific delta-endotoxin genes.

Main Methods:

  • Bioassays using Plagiodera versicolora larvae.
  • Agarose gel electrophoresis for plasmid pattern analysis.
  • Amino acid sequencing of crystal proteins.

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  • Genomic library construction and screening using a synthetic probe.
  • Restriction enzyme digestion (EcoRI) and analysis of recombinant plasmids.

    • Main Results:

    • Strain YM-03 exhibited the highest toxicity against P. versicolora.
    • Significant variations in plasmid patterns were observed among the five Bt strains.
    • Coleopteran-specific delta-endotoxin genes were identified in positive clones (LE392 (pBYM2), LE392 (pBYM3), LE392 (pBYM4)).
    • Clones LE392 (pBYM3) and LE392 (pBYM4) showed identical EcoRI digestion patterns and contained the target gene with differential expression.

    Conclusions:

    • The study identified potent coleopterancidal Bt strains with unique genetic profiles.
    • The coleoptera-specific delta-endotoxin gene was successfully cloned and characterized.
    • Differential expression of the delta-endotoxin gene suggests regulatory mechanisms influencing insecticidal activity.