Actinomycetospora aeridis sp. nov., Actinomycetospora flava sp. nov. and Actinomycetospora aurantiaca sp. nov., endophytic actinobacteria isolated from wild orchid (Aerides multiflora Roxb).

Three novel mycelium-forming actinobacteria, designated OC33-EN06T, OC33-EN07T, and OC33-EN08T, were isolated from wild orchid (Aerides multiflora Roxb), collected from a hill evergreen forest in Northern Thailand. Strains OC33-EN06T and OC33-EN07T showed the highest 16S rRNA gene similarity with Actinomycetospora lutea TT00-04T, 99.17 and 99.45%, respectively. Strain OC33-EN08T showed high similarity with four species, namely 'Actinomycetospora termitidis Odt1-22T' (99.37%), Actinomycetospora chiangmaiensis DSM 45062T (99.02%), Actinomycetospora corticicola 014-5T (99.02%), and Actinomycetospora soli SF1T (98.81%). Comparative genome analysis of OC33-EN06T, OC33-EN07T, and OC33-EN08T with the closely related type strains showed that average nucleotide identity (ANI) based on blast, ANI based on MUMmer, and average amino acid identity values were less than 95% and the digital DNA-DNA hybridization values were less than 70%, all below the thresholds for species demarcation. The digital G+C content of OC33-EN06T, OC33-EN07T, and OC33-EN08T were 74.5, 74, and 74 mol%, respectively. These three strains developed bud-like chains of non-motile cylindrical spores with a smooth surface. The cell-wall peptidoglycan contained meso-diaminopimelic acid. The whole-cell sugars contained ribose, arabinose, and galactose. The predominant menaquinone was MK-8(H4). The phospholipid profile included phosphatidylcholine, phosphatidylethanolamine, phosphatidylmethylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, and phosphatidylinositol. Based on comparative analysis of genotypic, phenotypic and chemotaxonomic data, strains OC33-EN06T (=TBRC 18349T=NBRC 116543T), OC33-EN07T (=TBRC 18350T=NBRC 116544T), and OC33-EN08T (=TBRC 18318T=NBRC 116542T) represent the type strains of three novel species of the genus Actinomycetospora for which the names Actinomycetospora aeridis sp. nov., Actinomycetospora flava sp. nov., and Actinomycetospora aurantiaca sp. nov., are proposed.

Glycomyces amatae sp. nov., isolated from a yellow-ringed grass moth (Amata sperbius).

A novel mycelium-forming actinomycete strain, designated A-F 0318T, was isolated from a yellow-ringed grass moth (Amata sperbius) collected from Phitsanulok Province, Thailand. Long chains of non-motile cylindrical spores with a smooth surface developed on aerial mycelia. The polyphasic taxonomic study suggested that strain A-F 0318T belonged to the genus Glycomyces. The 16S rRNA gene sequence analysis indicated that strain A-F 0318T was closely related to Glycomyces harbinensis LL-DO5139T with 97.94 % sequence similarity. The average nucleotide identity (ANI) based on blast, ANI based on the MUMmer algorithm and average amino acid identity values of strain A-F 0318T with G. harbinensis LL-DO5139T were 86.9, 89.1 and 84.24 %, respectively. The digital DNA-DNA hybridization value between A-F 0318T and its closest relative, G. harbinensis LL-DO5139T was 33.8 %. The digital G+C content of the genomic DNA was 71.7 mol%. The cell-wall peptidoglycan contained meso-diaminopimelic acid. The whole-cell sugars contained ribose, xylose, glucose and galactose. The predominant menaquinone was MK-10(H4). The predominant fatty acids were iso-C16 : 0, anteiso-C15 : 0, anteiso-C17 : 0 and iso-C16 : 1 G. The polar lipid profile consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, phosphatidylinositol mannoside, two unknown phosphoglycolipids and one unknown phospholipid. Based on comparative analysis of genotypic, phenotypic and chemotaxonomic data, the novel actinomycete strain A-F 0318T (=TBRC 13612T=NBRC 115417T) represents the type strain of a novel species, for which the name Glycomyces amatae sp. nov. is proposed.

Prauserella cavernicola sp. nov., isolated from cave rock.

A novel actinomycete, designated strain ASG 168T, was isolated from cave rock collected from Stegodon Sea Cave in Thailand. Long chains of non-motile spores that were oval or spherical in shape with a smooth surface developed on aerial mycelia. Substrate mycelia fragmented into irregular rod-shaped elements. A polyphasic taxonomic study showed that strain ASG 168T had typical characteristics of members of the genus Prauserella. 16S rRNA gene sequence analysis indicated that strain ASG 168T shared 97.5 % similarity with Prauserella marina MS498T and 96.7 % with Prauserella coralliicola SCSIO 11529T. Average nucleotide identity values with P. coralliicola SCSIO 11529T and P. marina MS498T were 82.98 and 76.08 %, respectively. The cell-wall peptidoglycan contained meso-diaminopimelic acid. The whole-cell sugars contained ribose, arabinose and galactose. The predominant menaquinone was MK-9(H4). The predominant fatty acids were iso-C16 : 0, C16 : 0 and summed feature 3 (C16 : 1 ω7c/C16 : 1 ω6c). The phospholipid profile consisted of phosphatidylethanolamine, phosphatidylmethylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, phosphatidylinositol mannosides and two unknown phospholipids. The G+C content of the genomic DNA was 70.6 mol%. Differentiation of strain ASG 168T from closely related species was evident from digital DNA-DNA hybridization values of 29.2 and 21.3 % with P. coralliicola and P. marina, respectively. Based on comparative analysis of phenotypic, chemotaxonomic and genotypic data, the novel actinomycete strain ASG 168T (=TBRC 13679T=NBRC 114887T) is proposed to be the type strain of a novel species, Prauserella cavernicola sp. nov.

Cryptosporangium phraense sp. nov., isolated from plant litter.

A novel zoosporic actinomycete, designated strain A-T 5661T, was isolated from plant litter collected in Thailand. Strain A-T 5661T developed large multilocular sporangia containing motile sporangiospores at the ends of sporangiophores on substrate mycelium. The results of our polyphasic taxonomic study demonstrated that the strain had characteristics typical of members of the genus Cryptosporangium. The 16S rRNA gene sequence and phylogenetic analyses indicated that strain A-T 5661T shared the sequence similarity ≤98.5 % with all members of the genus Cryptosporangium. The values of DNA-DNA relatedness that distinguished this novel strain from its closest related species were below 70 %. The cell-wall peptidoglycan contained meso-diaminopimelic acid. The whole-cell sugars were acofriose, arabinose, galactose, glucose, mannose, xylose and ribose. The predominant menaquinones were MK-9(H8) and MK-9(H6) with minor amounts of MK-9(H4) and MK-9(H2). The predominant fatty acids were iso-C16 : 0, C18 : 1ω9c and anteiso-C17 : 0. The polar lipid profile consisted of phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, phosphatidylinositol mannosides and unidentified polar lipids. The G+C content of the genomic DNA was 71.6 mol%. Based on comparative analysis of phenotypic, chemotaxonomic and genotypic data, the novel zoosporic actinomycete A-T 5661T (=TBRC 8127T=NBRC 113237T) is proposed to be the type strain of a novel species, Cryptosporangium phraense sp. nov.

Gordonia asplenii sp. nov., isolated from humic soil on bird's nest fern (Asplenium nidus L.).

A novel member of the actinobacteria, designated strain A-T 0013T, was isolated from humic soil on a bird's nest fern (Asplenium nidus L.) collected from Khao Yai National Park in Thailand. According the results of a polyphasic taxonomic study, A-T 0013T had characteristics typical of members of the genus Gordonia. The 16S rRNA gene sequence indicated that A-T 0013T shared ≤98 % sequence similarity with all members of the genus Gordonia. The most closely related species was Gordonia effusa IFM 10200T (97.92 % sequence similarity). The average nucleotide identity based on blast (ANIb) value with G. effusa IFM 10200T was 76.81 %. The cell-wall peptidoglycan contained meso-diaminopimelic acid. The whole-cell sugars contained ribose, arabinose and galactose. The predominant menaquinone was MK-9(H2). The predominant fatty acids were C16 : 0, C18 : 1ω9c, summed feature 3 (C16 : 1ω7c/C16 : 1ω6c), and C18 : 0 10-methyl. Mycolic acid was present. The polar lipid profile for this strain ncluded diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and phosphatidylinositol mannosides. The G+C content of the genomic DNA was 66.1 mol%. Differentiation of A-T 0013T from the most closely related species, Gordonia effusa IFM 10200T, was evident from digital DNA-DNA hybridization values of 21.8 %. On the basis of the results of comparative analysis of phenotypic, chemotaxonomic and genotypic data, strain A-T 0013T is concluded to represent a novel species of the genus Gordonia, for which the name Gordonia asplenii sp. nov. is proposed. The type strain is A-T 0013T (=TBRC 11910T=NBRC 114549T).

Verrucosispora endophytica sp. nov., isolated from the root of wild orchid (Grosourdya appendiculata (Blume) Rchb.f.).

A novel endophytic filamentous bacterium strain A-T 7972T, was isolated from wild orchid Grosourdya appendiculata (Blume) Rchb.f. collected in Thailand. The bacterium developed single non-motile spores with warty surface on substrate mycelia. The taxonomic position was described using a polyphasic approach. The 16S rRNA gene sequence and phylogenetic analysis indicated that strain A-T 7972T belonged to the genus Verrucosispora and shared the highest sequence similarity with V. lutea YIM 013T (98.71 %) and V. gifhornensis DSM 44337T (98.53 %). The values of DNA-DNA relatedness that distinguished this novel strain from its closest species were below 70 %. The cell-wall peptidoglycan contained meso-diaminopimelic acid. The whole-cell sugars were mannose, ribose, glucose and xylose. The predominant menaquinone was MK-9(H4). The predominant fatty acids were branched fatty acids iso-C15 : 0 and iso-C16 : 0. The diagnostic phospholipids profile consisted of phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylinositol and phosphatidylinositol mannosides. The G+C content of the genomic DNA was 70.5 mol%. Based on its phenotypic, chemotaxonomic and genotypic characteristics, the new isolate A-T 7972T (=BCC 50981T=TBRC 6031T=NBRC 112512T) is proposed to be the type strain of novel species, Verrucosispora endophytica sp. nov.