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Applied and Environmental Microbiology, September 1999, p. 4032-4039, Vol. 65, No. 9
Unité de Biochimie
Microbienne1 and Laboratoire des
Fermentations,
Received 4 March 1999/Accepted 8 July 1999
The main problems with Bacillus thuringiensis products
for pest control are their often narrow activity spectrum, high
sensitivity to UV degradation, and low cost effectiveness (high potency
required). We constructed a sporulation-deficient SigK
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Development and Field Performance of a Broad-Spectrum Nonviable
Asporogenic Recombinant Strain of Bacillus thuringiensis
with Greater Potency and UV Resistance


B. thuringiensis strain that expressed a chimeric
cry1C/Ab gene, the product of which had high activity
against various lepidopteran pests, including Spodoptera
littoralis (Egyptian cotton leaf worm) and Spodoptera
exigua (lesser [beet] armyworm), which are not readily
controlled by other Cry
-endotoxins. The SigK
host
strain carried the cry1Ac gene, the product of which is highly active against the larvae of the major pests Ostrinia
nubilalis (European corn borer) and Heliothis
virescens (tobacco budworm). This new strain had greater potency
and a broader activity spectrum than the parent strain. The crystals
produced by the asporogenic strain remained encapsulated within the
cells, which protected them from UV degradation. The
cry1C/Ab gene was introduced into the B. thuringiensis host via a site-specific recombination vector so
that unwanted DNA was eliminated. Therefore, the final construct contained no sequences of non-B. thuringiensis origin. As
the recombinant strain is a mutant blocked at late sporulation, it does
not produce viable spores and therefore cannot compete with wild-type
B. thuringiensis strains in the environment. It is thus a
very safe biopesticide. In field trials, this new recombinant strain
protected cabbage and broccoli against a pest complex under natural
infestation conditions.
*
Corresponding author. Mailing address: Unité de
Biochimie Microbienne, Institut Pasteur, 25-28, rue du Docteur Roux,
75724 Paris Cedex 15, France. Phone: 33 1 45 68 88 12. Fax: 33 1 45 68 89 38. E-mail: vsanchis{at}pasteur.fr.
Present address: Station de Recherches de Lutte Biologique, INRA,
La Minière, 78285 Guyancourt, France.
Present address: Departamento de Biologia Geral, Centro de
Ciencias Biologicas, Universidade Estadual de Londrina, Campus Universitario, 86051-970 Londrina, Pr., Brasil.
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