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Appl. Environ. Microbiol. doi:10.1128/AEM.02678-06
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Clonothrix fusca (Roze, 1896), a Filamentous, Sheathed, Methanotrophic -Proteobacterium

Dipartimento di Scienze e Tecnologie Biologiche ed Ambientali, Università degli Studi di Lecce, Via Monteroni, 73100 Lecce, Dipartimento di Scienze Ambientali, Seconda Università degli Studi di Napoli, Via A. Vivaldi 43,81100, Caserta, Istituto di Genetica e Biofisica "Adriano Buzzati-Traverso", Consiglio Nazionale delle Ricerche, via Pietro Castellino 111, 80131 Napoli, Istituto Motori, Consiglio Nazionale delle Ricerche, Viale Marconi 8, 80125 Napoli, Italy

^* To whom correspondence should be addressed. Email: alifano{at}ilenic.unile.it.

	Abstract

Crenothrix polyspora (Cohn, 1870) and Clonothrix fusca (Roze, 1896) are two filamentous, sheathed microorganisms exhibiting complex morphological differentiation, whose phylogeny and physiology have been obscure for a long time due to inability to cultivate them. Very recently, DNA sequencing data from uncultured C. polyspora-enriched material have suggested that Crenothrix is a methane oxidizing -Proteobacterium (Stoecker, K., B. Bendinger, B. Schoning, P. H. Nielsen, J. L. Nielsen, C. Baranyi, E. R. Toenshoff, H. Daims, and M. Wagner. 2006. Proc. Natl. Acad. Sci. U S A. 103:2363-2367). In contrast, the possible ecological function of C. fusca, originally considered a developmental stage of C. polyspora, is unknown. In this study temporal succession of two filamentous, sheathed microorganisms resembling Cohn's Crenothrix and Roze's Clonothrix was observed by analyzing the microbial community of an artesian well by optical microscopy. Combined culture-based and culture-independent approaches enabled us to assign C. fusca to a novel subgroup of methane oxidizing -Proteobacteria distinct from that of C. polyspora. This assignment was supported by i.) methane uptake and assimilation experiments, ii.) ultrastructural data showing the presence in C. fusca cytoplasm of an elaborate membrane system resembling that of methanotrophic -Proteobacteria, and iii.) sequencing data demonstrating the presence in its genome of a methanol dehydrogenase subunit-encoding gene (mxaF) and a conventional particulate methane mono-oxygenase subunit-encoding gene (pmoA) different from the unusual pmoA (u-pmoA) of C. polyspora.

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