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Numerical taxonomic, DNA-DNA hybridization, and phospholipid fatty acid composition analyses were performed on an extensive range of methanotrophic strains, including reference strains and environmental isolates obtained from sites throughout eastern Australia. When the results of these studies were related to the results of a study based on genomic physicochemical properties, they clarified group I and I1 methanotroph genus and species interrelationships. The group I methanotrophs were found to be made up of three broadly phenotypically and genotypically homologous clusters of species. The first group I methanotroph cluster included the carotenoid-containing species Methylomonas methanica, Methylomonas fodinarum, and Methy- lomonas aurantiucu. These species represent the true members of the genus Methylomonas. The second group I methanotroph cluster was made up of two subclusters of strains. One subcluster included species not capable of producing resting cells and consisted of the species "Methylomonas ugile," "Methylomonas alba," and Methylomonas pelagica. The other subcluster included species capable of forming desiccation-resistant cysts and included Methylococcus luteus, marine Methylomonas-like strains, and Methylococcus whittenburyi. Strains designated "Methylococcus ucrainicus" and Methylococcus vinelandii were found to be synonyms of Methylo- coccus whittenburyi, while Methylococcus bovis was a synonym of Methylococcus luteus. It is proposed that these subclusters represent a new genus, Methylobacter gen. nov. The species in the new genus are type species Methylobacter luteus comb. nov., Methylobacter agilis sp. nov., Methylobacter albus sp. nov., nom. rev., Methylobacter marinus sp. nov., Methylobacter pelagicus comb. nov., and Methylobacter whittenburyi comb. nov. The remaining group I methanotrophs included the moderately thermophilic species Methylococcus capsulatus and Methylococcus thermophilus and a group of unnamed strains closely related to Methylococcus capsulatus. It is proposed that these species represent the true members of the genus Methylococcus. The group I1 methanotrophs consisted of two closely related groups. The first group included budding, exospore- producing strains, while the second group included nonmotile, cyst-forming strains. These groups represent the genera Methylosinus and Methyocystis, which are revived here. The genus Methylosinus gen. nov., nom. rev. includes the species Methylosinus trichosporium sp. nov., nom. rev. and Methylosinus sporium sp. nov., nom. rev., while the genus Methylocystis gen. nov., nom. rev. includes the species Methylocystis parvus sp. nov., nom. rev. and Methylocystis echinoides sp. nov., nom. rev. The methanotrophs represent a subset of gram-negative, obligately methylotrophic bacteria that utilize methane as their primary source of carbon and energy. The difficulty in obtaining pure cultures, the lack of discriminating pheno- typic characteristics, and transgressions against the Interna- tional Code of Nomenclature of Bacteria (30, 56) have resulted in classification problems in methanotroph taxon- omy. The methanotrophs can be separated into two assem- blages, which are usually referred to as types or groups I and I1 (56). To avoid confusion with such concepts as type strain, type species, etc., in this study we used the terms group I and group 11. The group I and I1 methanotrophs differ in their intracytoplasmic membrane ultrastructures, carbon assimi-

international journal of systematic bacteriology

Revised Taxonomy of the Methanotrophs: Description of Methylobacter gen. nov., Emendation of Methylococcus, Validation of Methylosinus and Methylocystis Species, and a Proposal that the Family Methylococcaceae Includes Only the Group I Methanotrophs