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Anaerolinea thermolimosa sp. nov., Levilinea saccharolytica gen. nov., sp. nov. and Leptolinea tardivitalis gen. nov., sp. nov., novel filamentous anaerobes, and description of the new classes Anaerolineae classis nov. and Caldilineae classis nov. in the bacterial phylum Chloroflexi

¹ Institute for Biological Resources and Functions, National Institute of Advanced Industrial Science and Technology (AIST), Higashi 1-1-1, Tsukuba, Ibaraki 305-8566, Japan
² Department of Environmental Systems Engineering, Nagaoka University of Technology, Nagaoka, Niigata 940-2188, Japan

Correspondence
Yuji Sekiguchi
y.sekiguchi{at}aist.go.jp

One thermophilic (strain IMO-1^T) and two mesophilic (strains KIBI-1^T and YMTK-2^T) non-spore-forming, non-motile, Gram-negative, multicellular filamentous micro-organisms, which were previously isolated as members of the tentatively named class ‘Anaerolineae’ of the phylum Chloroflexi, were characterized. All isolates were strictly anaerobic micro-organisms. The length of the three filamentous isolates was greater than 100 µm and the width was 0.3–0.4 µm for strain IMO-1^T, 0.4–0.5 µm for strain KIBI-1^T and thinner than 0.2 µm for strain YMTK-2^T. Strain IMO-1^T could grow at pH 6.0–7.5 (optimum growth at pH 7.0). The optimal temperature for growth of strain IMO-1^T was around 50 °C (growth occurred between 42 and 55 °C). Growth of the mesophilic strains KIBI-1^T and YMTK-2^T occurred at pH 6.0–7.2 with optimal growth at pH 7.0. Both of the mesophilic strains were able to grow in a temperature range of 25–50 °C with optimal growth at around 37 °C. Yeast extract was required for growth of all three strains. All the strains could grow with a number of carbohydrates in the presence of yeast extract. The G+C contents of the DNA of strains IMO-1^T, KIBI-1^T and YMTK-2^T were respectively 53.3, 59.5 and 48.2 mol%. Major fatty acids for thermophilic strain IMO-1^T were anteiso-C_{17 : 0}, iso-C_{15 : 0}, C_{16 : 0} and anteiso-C_{15 : 0}, whereas those for mesophilic strains KIBI-1^T and YMTK-2^T were branched C_{14 : 0}, iso-C_{15 : 0}, C_{16 : 0} and branched C_{17 : 0}, and branched C_{17 : 0}, C_{16 : 0}, C_{14 : 0} and C_{17 : 0}, respectively. Detailed phylogenetic analyses based on their 16S rRNA gene sequences indicated that the isolates belong to the class-level taxon ‘Anaerolineae’ of the bacterial phylum Chloroflexi, which for a long time had been considered as a typical uncultured clone cluster. Their morphological, physiological, chemotaxonomic and genetic traits strongly support the conclusion that these strains should be described as three novel independent taxa in the phylum Chloroflexi. Here, Anaerolinea thermolimosa sp. nov. (type strain IMO-1^T=JCM 12577^T=DSM 16554^T), Levilinea saccharolytica gen. nov., sp. nov. (type strain KIBI-1^T=JCM 12578^T=DSM 16555^T) and Leptolinea tardivitalis gen. nov., sp. nov. (type strain YMTK-2^T=JCM 12579^T=DSM 16556^T) are proposed. In addition, we formally propose to subdivide the tentative class-level taxon ‘Anaerolineae’ into Anaerolineae classis nov. and Caldilineae classis nov. We also propose the subordinate taxa Anaerolineales ord. nov., Caldilineales ord. nov., Anaerolineaceae fam. nov. and Caldilineaceae fam. nov.

The GenBank/EMBL/DDBJ accession numbers for the 16S rRNA gene sequences of strains IMO-1^T, YMTK-2^T and KIBI-1^T are AB109437–AB109439.

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