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Gene disruption and characterization of a class V chitin synthase in Botrytis cinerea

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Cell-wall chitin biosynthesis in the phytopathogenic fungus Botrytis cinerea involves 7 classes of chitin synthases. In this study, we disrupted a gene encoding a chitin synthase with a myosin-like motor domain (BcchsV) through Agrobacterium tumefaciens mediated transformation. The resulting mutant was not significantly affected in either growth characteristics or pathogenicity on tomato leaves. Surprisingly, the BcchsV mutant exhibited a 31% (m/m) increase in its chitin content compared with the wild-type strain. In addition, the BcchsV mutant showed increased sensitivity to Calcofluor White and slightly enhanced tolerance to cell-wall disturbing substances and osmosis regulators, including SDS, sorbitol, and NaCl. These results suggest that Bcchs does not play an essential role in the synthesis of cell-wall chitin in B. cinerea. However, disruption of this gene provoked a compensatory mechanism regulating the cellular response to cell-wall damage.

La biosynthèse de la chitine de la paroi cellulaire du champignon phytopathogène Botrytis cinerea implique 7 classes de chitine synthases. Dans cette étude, nous avons interrompu le gène codant une chitine synthase (BcchsV) avec un domaine moteur de type myosine via une transformation réalisée par Agrobacterium tumefaciens. Le mutant qui en a résulté n’était pas significativement affecté quant à ses caractéristiques de croissance et sa pathogénicité sur des feuilles de tomates. Étonnamment, le contenu en chitine du mutant BcchsV était plus élevé de 31 % (p/p) comparativement à la souche sauvage. Conjointement à ce changement, le mutant Bcchs était plus sensible au calcofluor blanc et légèrement plus tolérant aux substances perturbant la membrane ou les substances osmorégulatrices, notamment le SDS, le sorbitol et le NaCl. Ces résultats suggèrent que le gène BcchsV ne joue pas un rôle essentiel dans la synthèse de la chitine de la paroi cellulaire du B. cinerea. Cependant, l’interruption de ce gène provoque un mécanisme compensateur qui régule la réponse cellulaire aux dommages de la paroi cellulaire.

Document Type: Research Article

Publication date: November 1, 2009

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  • Published since 1954, this monthly journal contains new research in the field of microbiology including applied microbiology and biotechnology; microbial structure and function; fungi and other eucaryotic protists; infection and immunity; microbial ecology; physiology, metabolism and enzymology; and virology, genetics, and molecular biology. It also publishes review articles and notes on an occasional basis, contributed by recognized scientists worldwide.
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