We isolated from compost an aerobic, thermophilic, Gram-stain-positive, spore-forming bacterium that formed branched vegetative and aerial mycelia. This strain, designated SK20-1T, grew at 31–58 °C, with optimum growth at 50 °C, while no growth was observed below 28 or above 60 °C. The pH range for growth was 5.4–8.7, with optimum growth at pH 7.0, while no growth was observed below pH 5.0 or above pH 9.1. Strain SK20-1T was able to hydrolyse polysaccharides such as cellulose, xylan and chitin. The DNA G+C content was 54.0 mol%. The major fatty acid was iso-C17 : 0 and the major menaquinone was MK-9(H2). The cell wall contained glutamic acid, serine, alanine and ornithine in a molar ratio of 1.00 : 1.07 : 2.64 : 0.83. The polar lipids consisted of phosphatidylinositol, phosphatidylinositol mannosides, phosphatidylglycerol, diphosphatidylglycerol and an unknown glycolipid. Cell-wall sugars were rhamnose and mannose. Detailed phylogenetic analysis based on 16S rRNA gene sequences indicated that strain SK20-1T belongs to the class Ktedonobacteria, and that the strain is most closely related to Ktedonobacter racemifer SOSP1-21T (88.5 %). On the basis of its phenotypic features and phylogenetic position, we propose that SK20-1T represents a novel genus and species, Thermosporothrix hazakensis gen. nov., sp. nov., within the new family Thermosporotrichaceae fam. nov. The type strain of Thermosporothrix hazakensis is strain SK20-1T (=JCM 16142T =ATCC BAA-1881T). In addition, we propose an emended description of the class Ktedonobacteria to classify the class in the phylum Chloroflexi.

The prevalence of antibiotic resistance and the decrease in novel antibiotic discovery in recent years necessitates the identification of potentially novel microbial resources to produce natural products. Ktedonobacteria , a class of deeply branched bacterial lineage in the ancient phylum Chloroflexi , are ubiquitous in terrestrial environments and characterized by their large genome size and complex life cycle. These characteristics indicate Ktedonobacteria as a potential active producer of bioactive compounds. In this study, we observed the existence of a putative “megaplasmid,” multiple copies of ribosomal RNA operons, and high ratio of hypothetical proteins with unknown functions in the class Ktedonobacteria . Furthermore, a total of 104 antiSMASH-predicted putative biosynthetic gene clusters (BGCs) for secondary metabolites with high novelty and diversity were identified in nine Ktedonobacteria genomes. Our investigation of domain composition and organization of the non-ribosomal peptide synthetase and polyketide synthase BGCs further supports the concept that class Ktedonobacteria may produce compounds structurally different from known natural products. Furthermore, screening of bioactive compounds from representative Ktedonobacteria strains resulted in the identification of broad antimicrobial activities against both Gram-positive and Gram-negative tested bacterial strains. Based on these findings, we propose the ancient, ubiquitous, and spore-forming Ktedonobacteria as a versatile and promising microbial resource for natural product discovery.

Two thermophilic, Gram-stain-positive, sporulating bacterial strains, which formed branched vegetative and aerial mycelia, were isolated from fallen leaves sampled from geothermal soils and designated ONI-1T and ONI-5T. Strain ONI-1T grew at 50–74 °C, with optimum growth at 60–65 °C, and strain ONI-5T grew at 45–74 °C, with optimum growth at 60–65 °C. The pH range for growth of the strains was pH 4.6–8.0, with optimum growth at pH 7.0. The DNA G+C contents of strains ONI-1T and ONI-5T were 60.2 and 58.1 mol%, respectively. The major fatty acid was iso-C17 : 0 and the major menaquinone was MK-9(H2). The cell walls of the strains contained glutamic acid, serine, glycine, histidine, alanine and ornithine. The polar lipids consisted of phosphatidylinositol, phosphatidylglycerol and a glycolipid. The cell-wall sugar was rhamnose. Detailed phylogenetic analysis based on 16S rRNA gene sequences indicated that the strains belong to the class Ktedonobacteria and that strains ONI-1T and ONI-5T are most closely related to Thermosporothrix hazakensis SK20-1T (85.3 and 84.5 % sequence similarity, respectively). 16S rRNA gene sequence similarity between the two strains was 96.6 %. Based on the phenotypic features and phylogenetic position, we propose that strains ONI-1T and ONI-5T constitute a novel genus containing two novel species, for which we propose the names Thermogemmatispora onikobensis gen. nov., sp. nov. (the type species; type strain ONI-1T  = JCM 16817T  = KCTC 19768T) and Thermogemmatispora foliorum sp. nov. (type strain ONI-5T  = JCM 16818T  = KCTC 19767T), within the new family Thermogemmatisporaceae fam. nov. and order Thermogemmatisporales ord. nov.

A mesophilic, Gram-stain-positive, spore-forming bacterium that formed branched mycelia was isolated from paddy soil in Gunung Salak (Mount Salak), West Java, Indonesia. This strain, designated S-27T, grew at temperatures between 20 and 37 °C; the optimum growth temperature was 25 to 30 °C, and no growth was observed at 15 or 45 °C. The pH range for growth was pH 3.5 to 8.6; the optimum pH was 6.0, and no growth was observed at pH 3.0 or 9.2. Strain S-27T was able to hydrolyse polysaccharides such as starch, cellulose and xylan. The G+C content of the DNA of strain S-27T was 55.7 mol%. The major fatty acids were iso-C17 : 0 and C16 : 1 2-OH, and the major menaquinone was MK-9 (H2). The cell wall of strain S-27T contained d-glutamic acid, glycine, l-alanine, d-alanine, l-ornithine and β-alanine in a molar ratio of 1.0 : 1.6 : 1.4 : 0.6 : 0.9 : 1.1. The polar lipids consisted of phosphatidylglycerol, phosphatidylinositol and two glycolipids. The major cell-wall sugar was arabinose. Detailed phylogenetic analysis based on 16S rRNA gene sequences indicated that strain S-27T belongs to the order Ktedonobacterales and is most closely related to Ktedonobacter racemifer SOSP1-21T (89.6 % sequence identity). On the basis of its chemotaxonomic and phenotypic features and phylogenetic position, we concluded that strain S-27T represents a novel genus and species, for which we propose the name Dictyobacter aurantiacus gen. nov., sp. nov. The type strain of Dictyobacter aurantiacus is strain S-27T (=NBRC 109595T=InaCC B312T). Emendation of the description of the genus Thermosporothrix is also provided.

Bacteria with an actinomycetes-like morphology have recently been discovered, and the class Ktedonobacteria was created for these bacteria in the phylum Chloroflexi. They may prove to be a valuable resource with the potential to produce unprecedented secondary metabolites. However, our understanding of their diversity, richness, habitat, and ecological significance is very limited. We herein developed a 16S rRNA gene-targeted, Ktedonobacteria-specific primer and analyzed ktedonobacterial amplicons. We investigated abundance, diversity, and community structure in forest and garden soils, sand, bark, geothermal sediment, and compost. Forest soils had the highest diversity among the samples tested (1181–2934 operational taxonomic units [OTUs]; Chao 1 estimate, 2503–5613; Shannon index, 4.21–6.42). A phylogenetic analysis of representative OTUs revealed at least eight groups within unclassified Ktedonobacterales, expanding the known diversity of this order. Ktedonobacterial communities markedly varied among our samples. The common mesic environments (soil, sand, and bark) were dominated by diverse phylotypes within the eight groups. In contrast, compost and geothermal sediment samples were dominated by known ktedonobacterial families (Thermosporotrichaceae and Thermogemmatisporaceae, respectively). The relative abundance of Ktedonobacteria in the communities, based on universal primers, was ≤0.8%, but was 12.9% in the geothermal sediment. These results suggest that unknown diverse Ktedonobacteria inhabit common environments including forests, gardens, and sand at low abundances, as well as extreme environments such as geothermal areas.

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A Gram-stain-positive thermophilic bacterium, designated strain Nis3T, was isolated from compost. The strain grew at 23–57 °C (optimum, 50 °C); no growth was observed below 15 or above 60 °C. The pH range for growth was 5.9–8.8 (optimum, 7.0); no growth was observed below pH 5.4 or above pH 9.3. The DNA G+C content of strain Nis3T was 63.4 mol%. The dominant quinone type was ubiquinone Q-10. The major fatty acids were C18 : 1ω7c, C19 : 0ω8c cyclo and C18 : 0. The polar lipids comprised phosphatidylcholine, phosphatidylglycerol, phosphatidylethanolamine, hydroxyphosphatidylethanolamine, phosphatidylinositol, phosphatidylmonomethylethanolamine, an unknown glycolipid and a ninhydrin-positive phospholipid. 16S rRNA gene sequence analysis assigned this bacterium to the family Phyllobacteriaceae in the Alphaproteobacteria but it shared less than 95.2 % sequence similarity with other members of the family. The chemotaxonomic and phenotypic characteristics of strain Nis3T differed in some respects from those of members of the family Phyllobacteriaceae . Therefore, strain Nis3T is considered to represent a novel species of a new genus in the family Phyllobacteriaceae , for which the name Thermovum composti gen. nov., sp. nov. is proposed. The type strain is Nis3T ( = JCM 17863T = KCTC 23707T).

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