Novel isolate of Bacillus thuringiensis subsp. thuringiensis that produces a quasicuboidal crystal of Cry1Ab21 toxic to larvae of Trichoplusia ni

Appl Environ Microbiol. 2008 Feb;74(4):923-30. doi: 10.1128/AEM.01955-07. Epub 2007 Dec 14.

Abstract

A new isolate (IS5056) of Bacillus thuringiensis subsp. thuringiensis that produces a novel variant of Cry1Ab, Cry1Ab21, was isolated from soil collected in northeastern Poland. Cry1Ab21 was composed of 1,155 amino acids and had a molecular mass of 130.5 kDa, and a single copy of the gene coding for this endotoxin was located on a approximately 75-kbp plasmid. When synthesized by the wild-type strain, Cry1Ab21 produced a unique, irregular, bipyramidal crystal whose long and short axes were both approximately 1 microm long, which gave it a cuboidal appearance in wet mount preparations. In diet incorporation bioassays, the 50% lethal concentrations of the crystal-spore complex were 16.9 and 29.7 microg ml(-1) for second- and fourth-instar larvae of the cabbage looper, Trichoplusia ni, respectively, but the isolate was essentially nontoxic to larvae of the beet armyworm, Spodoptera exigua. A bioassay of autoclaved spore-crystal preparations showed no evidence of beta-exotoxin activity, indicating that toxicity was due primarily to Cry1Ab21. Studies of the pathogenesis of isolate IS5056 in second-instar larvae of T. ni showed that after larval death the bacterium colonized and subsequently sporulated extensively throughout the cadaver, suggesting that other bacteria inhabiting the midgut lumen played little if any role in mortality. As T. ni is among the most destructive pests of vegetable crops in North America and has developed resistance to B. thuringiensis, this new isolate may have applied value.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacillus thuringiensis / genetics*
  • Bacillus thuringiensis Toxins
  • Bacterial Proteins / genetics*
  • Bacterial Proteins / metabolism
  • Bacterial Proteins / toxicity
  • Bacterial Toxins / genetics*
  • Bacterial Toxins / metabolism
  • Bacterial Toxins / toxicity
  • Base Sequence
  • Blotting, Southern
  • Cloning, Molecular
  • DNA Primers / genetics
  • Endotoxins / genetics*
  • Endotoxins / metabolism
  • Endotoxins / toxicity
  • Hemolysin Proteins / genetics*
  • Hemolysin Proteins / metabolism
  • Hemolysin Proteins / toxicity
  • Larva / drug effects
  • Molecular Sequence Data
  • Moths / drug effects
  • Poland
  • Sequence Analysis, DNA
  • Serotyping
  • Soil Microbiology*
  • Species Specificity

Substances

  • Bacillus thuringiensis Toxins
  • Bacterial Proteins
  • Bacterial Toxins
  • DNA Primers
  • Endotoxins
  • Hemolysin Proteins
  • insecticidal crystal protein, Bacillus Thuringiensis