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Microbiology 146 (2000), 2825-2832
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Microbiology (2000), 146, 2825-2832.
© 2000 Society for General Microbiology

Pathogenicity and Medical Microbiology

The plcR regulon is involved in the opportunistic properties of Bacillus thuringiensis and Bacillus cereus in mice and insects

Sylvie Salamitou1,2, Françoise Ramisse3, Michel Brehélin4, Denis Bourguet2, Nathalie Gilois2, Myriam Gominet1, Eric Hernandez5 and Didier Lereclus1,2

Unité de Biochimie Microbienne, CNRS (URA2172), Institut Pasteur, 25 rue du Docteur Roux, 75724 Paris cedex, France1
Unité de Lutte Biologique, INRA, La Minière, 78285 Guyancourt cedex, France2
Centre d’Etudes du Bouchet, Laboratoire de Microbiologie, 91710 Vert-Le-Petit, France3
Laboratoire de Pathologie Comparée, INRA-CNRS (URA 2209), Université Montpellier II, 34095 Montpellier, France4
Laboratoire de Biologie, HIA Bégin, 94160 Saint Mandé, France5

Author for correspondence: Didier Lereclus. Tel: +33 1 45 68 88 13. Fax: +33 1 45 68 89 38. e-mail: lereclus{at}pasteur.fr

Bacillus thuringiensis has been widely used for 40 years as a safe biopesticide for controlling agricultural pests and mosquitoes because it produces insecticidal crystal proteins. However, spores have also been shown to contribute to overall entomopathogenicity. Here, the opportunistic properties of acrystalliferous B. thuringiensis Cry- and Bacillus cereus strains were investigated in an insect species, Galleria mellonella, and in a mammal, BALB/c mice. In both animal models, the pathogenicity of the two bacterial species was similar. Mutant strains were constructed in which the plcR gene, encoding a pleiotropic regulator of extracellular factors, was disrupted. In larvae, co-ingestion of 106 spores of the parental strain with a sublethal concentration of Cry1C toxin caused 70% mortality whereas only 7% mortality was recorded if spores of the {Delta}plcR mutant strain were used. In mice, nasal instillation of 108 spores of the parental strain caused 100% mortality whereas instillation with the same number of {Delta}plcR strain spores caused much lower or no mortality. Similar effects were obtained if vegetative cells were used instead of spores. The cause of death is unknown and is unlikely to be due to actual growth of the bacteria in mice. The lesions caused by B. thuringiensis supernatant in infected mice suggested that haemolytic toxins were involved. The cytolytic properties of strains of B. thuringiensis and B. cereus, using sheep, horse and human erythrocytes and G. mellonella haemocytes, were therefore investigated. The level of cytolytic activity is highly reduced in {Delta}plcR strains. Together, the results indicate that the pathogenicity of B. thuringiensis strain 407 and B. cereus strain ATCC 14579 is controlled by PlcR.

Keywords: Bacillus, cytolysin, haemolysin, insect pathogen, plcR regulon




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