Expression regulation of the endochitinase-encoding gene sechi44 from the mycoparasite Stachybotrys elegans

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Abstract:

The regulation of the gene encoding the extracellular chitinase sechi44 produced by the mycoparasite Stachybotrys elegans was studied using real-time quantitative reverse-transcription polymerase chain reaction. Alteration of sechi44 expression was observed when S. elegans was in interaction with its host, Rhizoctonia solani, and also when the mycoparasite was grown on minimal media amended with different carbon and nitrogen sources. Direct contact with R. solani leading to mycoparasitism significantly up-regulated the expression of sechi44, although the analysis showed that sechi44 was constitutively expressed but at substantially lower levels. In addition, the study of sechi44 over 12 days showed that its expression followed a cyclical pattern with peaks every 2 days, which suggests that this gene has a role not only in mycoparasitism but also in growth. The addition of external carbon sources, such as N-acetylglucosamine, chitin, and R. solani cell wall (simulated mycoparasitism), triggered an increase in the expression of sechi44, which varied with time and carbon source. Among the carbon sources examined, N-acetylglucosamine induced the highest increase in sechi44 transcript levels. The addition of high concentrations of glucose and ammonium triggered a decrease of sechi44 expression, suggesting that sechi44 is subject to glucose and ammonium repression.Key words: mycoparasitism, Stachybotrys elegans, endochitinase-encoding gene, sechi44, real-time RT–PCR.

La régulation du gène codant la chitinase extracellulaire sechi44 produite par le mycoparasite Stachybotrys elegans fut étudiée à l'aide du réaction en chaîne de la polymérase à transcription inverse quantitatif. Un changement dans l'expression de sechi44 fut observé lorsque S. elegans était en interaction avec son hôte, Rhizoctonia solani, de même que lorsque le mycoparasite a été cultivé dans un milieu minimal enrichi avec différentes sources de carbone et d'azote. Un contact direct avec R. solani menant au mycoparasitisme a augmenté significativement l'expression de sechi44, bien que l'analyse ait démontré que sechi44 était exprimé constitutivement mais à des niveaux bien inférieurs. De plus, l'étude de sechi44 sur une période de 12 jours a démontré que son expression suivait un cycle avec des pics à chaque deux jours, ce qui indique que ce gène joue un rôle non seulement dans le mycoparasitisme mais également dans la croissance. L'ajout de sources de carbone externes comme le N-acétylglucosamine, la chitine et des parois cellulaires de R. solani (mycoparasitisme simulé) a déclenché une augmentation de l'expression de sechi44 qui variait selon le temps et la source de carbone. Parmi les sources de carbone examinées, la N-acétylglucosamine a induit la plus importante augmentation de la transcription de sechi44. L'ajout de concentrations élevées de glucose et d'ammonium a entraîné une diminution de l'expression de sechi44 indiquant que sechi44 est sujet à une répression par le glucose et l'ammonium.Mots clés : mycoparasitisme, Stachybotrys elegans, gène codant une endochitinase, sechi44, RT–PCR en temps réel.[Traduit par la Rédaction]

Document Type: Research Article

Publication date: November 1, 2006

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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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