Mikrobiol. Z. 2017; 79(2):3-12. Russian.
doi: https://doi.org/10.15407/microbiolj79.02.003

Phylogenetic Analysis of the Collection Strains of Methane Oxidizing Bacteria

Romanovskaya V.A.1, Belkova N.L.2, Parfenova V.V.2, Gladka G.V.1, Muchnyk Ph.V.1, Tashyrev O.B.1

1Zabolotny Institute of Microbiology and Virology, NAS of Ukraine
154 Akad. Zabolotny Str., Kyiv, 03143, Ukraine

2Limnological Institute SD, RAS
3 Ulan-Batorskaya Str., Irkutsk, 664033, Russia

Aim. Perform phylogenetic analysis to determine the taxonomic position of the collection strains of methane oxidizing bacteria (methanotrophs). Methods. Objects of research – 22 strains of obligate methanotrophs that were stored in the Ukrainian collection of microorganisms (UCM). Morphological and physiological properties of bacteria were studied by standard methods. Way methane assimilation established by determining in cell-free extracts of methanotrophs key enzymes of metabolism of C1 compounds. Phylogenetic position is determined construction of phylogenetic trees based on 16S rRNA genes methanotrophs showing the position of the strain among closely related and type species (software packages ClustalX 2.1, Mega v. 6.00) using the method of nearest neighbors. Results. The studied strains presented the families of Methylococcaceae (genus Methylobacter) and Methylocystaceae (genus Methylocystis and Methylosinus). According to phylogenetic and phenotypic analyzes they belong to the species Methylobacter marinus (5 strains), M. luteus (1 strain), Methylosinus trichosporium (3 strains), Methylocystis rosea (2 strains), M. hirsuta (1 strain). Nine strains of Methylobacter formed on the phylogenetic tree two separate clusters. These strains also differ by phenotypic characteristics from known species and, therefore, may represent new species. Conclusions. Phylogenetic and phenotypic analyzes have allowed to clarify the taxonomic status of collection methanotrophs strains.

Key words: obligate methane-oxidizing bacteria, genes 16S rRNA, phylogenetic analysis, the taxonomic position.

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