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Applied and Environmental Microbiology, February 2008, p. 923-930, Vol. 74, No. 4
0099-2240/08/$08.00+0     doi:10.1128/AEM.01955-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Novel Isolate of Bacillus thuringiensis subsp. thuringiensis That Produces a Quasicuboidal Crystal of Cry1Ab21 Toxic to Larvae of Trichoplusia ni

Department of Microbiology, Institute of Biology, University of Bialystok, 20B Swierkowa Street, PL15-950 Bialystok, Poland,¹ Department of Entomology, University of California, Riverside, Riverside, California 92521,² Department of Natural and Mathematical Science, California Baptist University, Riverside, California 92504,³ Interdepartmental Graduate Programs in Genetics and Microbiology, University of California, Riverside, Riverside, California 92521⁴

Received 25 August 2007/ Accepted 27 November 2007

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 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 µm 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 µg 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 β-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.

Published ahead of print on 14 December 2007.