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Applied and Environmental Microbiology, September 1999, p. 4032-4039, Vol. 65, No. 9
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

Vincent Sanchis,1,2,* Michel Gohar,3,dagger Josette Chaufaux,2 Olivia Arantes,1,Dagger Alain Meier,4 Hervé Agaisse,1,2 Jane Cayley,5 and Didier Lereclus1,2

Unité de Biochimie Microbienne1 and Laboratoire des Fermentations, Unité de Physiologie Cellulaire,4 Institut Pasteur, Centre National de la Recherche Scientifique, 75724 Paris Cedex 15, Station de Recherches de Lutte Biologique, Institut National de la Recherche Agronomique, La Minière, 78285 Guyancourt,2 and AgrEvo Prodetech, 13367 Marseille Cedex 11,3 France, and Hoechst Schering AgrEvo GmbH, Hoechst Works, D-65926 Frankfurt am Main, Germany5

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- 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 delta -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.

dagger Present address: Station de Recherches de Lutte Biologique, INRA, La Minière, 78285 Guyancourt, France.

Dagger Present address: Departamento de Biologia Geral, Centro de Ciencias Biologicas, Universidade Estadual de Londrina, Campus Universitario, 86051-970 Londrina, Pr., Brasil.


Applied and Environmental Microbiology, September 1999, p. 4032-4039, Vol. 65, No. 9
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.






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