Actinoallomurus soli sp. nov. and Actinoallomurus rhizosphaericola sp. nov., two novel actinobacteria isolated from rhizosphere soil of Oryza rufipogon Griff.

The taxonomic position of two novel Actinoallomurus strains isolated from rhizosphere soil of wild rice (Oryza rufipogon Griff.) was established using a polyphasic approach. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strains WRP6H-15T and WRP9H-5T were closely related to Actinoallomurus spadix JCM 3146T and Actinoallomurus purpureus TTN02-30T. Chemotaxonomic and morphological characteristics of both strains were consistent with members of the genus Actinoallomurus, while phenotypic properties, genome-based comparisons and phylogenomic analyses distinguished strains WRP6H-15T and WRP9H-5T from their closest phylogenetic relatives. The two strains showed nearly identical 16S rRNA gene sequences (99.9 %). Strain WRP6H-15T showed 68.7 % digital DNA-DNA hybridization, 95.9 % average nucleotide identity (ANI) based on blast and 96.4 % ANI based on MUMmer to strain WRP9H-5T. A phylogenomic tree based on draft genome sequences of the strains and representative of the genus Actinoallomurus confirmed the phylogenetic relationships. The genomes sizes of strains WRP6H-15T and WRP9H-5T were 9.42 Mb and 9.68 Mb, with DNA G+C contents of 71.5 and 71.3 mol%, respectively. In silico analysis predicted that the strains contain biosynthetic gene clusters encoding for specialized metabolites. Characterization based on chemotaxonomic, phylogenetic, phenotypic and genomic evidence demonstrated that strains WRP6H-15T and WRP9H-5T represent two novel species of the genus Actinoallomurus, for which the names Actinoallomurus soli sp. nov. (type strain WRP6H-15T=TBRC 15726T=NBRC 115556T) and Actinoallomurus rhizosphaericola sp. nov. (type strain WRP9H-5T=TBRC 15727T=NBRC 115557T) are proposed.

Saccharopolyspora oryzae sp. nov., isolated from rhizosphere soil of the wild rice species Oryza rufipogon.

A novel actinobacterium, designated as strain WRP15-2T, was isolated from rhizosphere soil of rice plant (Oryza rufipogon). The strain was Gram-stain-positive, aerobic, and non-motile. Phylogenetic analysis based on the 16S rRNA gene sequences showed that strain WRP15-2T fell into the genus Saccharopolyspora. The strain shared the highest 16S rRNA gene sequence similarity with the type strains Saccharopolyspora kobensis JCM 9109T (99.1%), Saccharopolyspora indica VRC122T (98.9%), and Saccharopolyspora antimicrobica DSM 45119T (98.7%). However, the digital DNA-DNA hybridization and average nucleotide identity values among these strains confirmed that the microorganism represented a novel member of the genus Saccharopolyspora. Chemotaxonomic data revealed that strain WRP15-2T possessed MK-9(H4) as the predominant menaquinone. It contained meso-diaminopimelic acid as the diagnostic diaminopimelic acid and arabinose, galactose, and ribose as predominant whole-cell sugars. The detected phospholipids were dominated by phosphatidylethanolamine, hydroxy-phosphatidylethanolamine, phosphatidylmethylethanolamine, hydroxy-phosphatidylmethylethanolamine, and phosphatidylcholine. The predominant cellular fatty acids were iso-C16:0, C16:0, and iso-C15:0. The G + C content of the genomic DNA was 69.5%. Based on these genotypic and phenotypic data, it is supported that strain WRP15-2T represents a novel species of the genus Saccharopolyspora, for which the name Saccharopolyspora oryzae sp. nov. is proposed. The type strain is WRP15-2T ( = TBRC 15728T = NBRC 115560T).

Streptomyces rhizoryzae sp. nov., isolated from paddy rhizosphere soil and formal proposal to reclassify Streptomyces albulus as a later heterotypic synonym of Streptomyces noursei.

The taxonomic position of a novel actinomycete, designated strain RS10V-4T, was determined using a polyphasic approach. Strain RS10V-4T was isolated from paddy rhizosphere soil of rice plant (Oryzae sativa L.). The morphological, physiological and chemotaxonomic properties were consistent with its classification in the genus Streptomyces. On the basis of 16S rRNA gene sequence analysis, strain RS10V-4T belongs to the genus Streptomyces and had the highest sequence similarity to Streptomyces noursei NBRC 15452T (98.3 %). The G+C content of the genomic DNA was 73.8 %. Digital DNA-DNA hybridization and average nucleotide identity values between the genome sequences of strain RS10V-4T and S.noursei ATCC 11455T were lower than the recommendation threshold values for the recognition of species within the same genus. The whole-cell hydrolysates of strain RS10V-4 T contained ll -diaminopimelic acid as the diagnostic diamino acid and the whole-cell sugars were glucose and ribose. The predominant menaquinones were MK-9(H6) and MK-9(H8). The predominant cellular fatty acids (>10 %) were iso-C16 : 0, anteiso-C15 : 0, iso-C14 : 0 and iso-C15 : 0. The polar lipids of strain RS10V-4T contained diphosphatidylglycerol, hydroxyphosphatidylethanolamine, phosphatidylethanolamine, phosphatidylinositol, an unidentified aminolipid, two unidentified lipids and an unidentified phospholipid. On the basis of these phenotypic and genotypic characteristics, it is supported that strain RS10V-4T represents a novel species of the genus Streptomyces, for which the name Streptomyces rhizoryzae sp. nov. is proposed. The type strain is RS10V-4T (=TBRC 15167T=NBRC 115345T). In addition, the comparison of the whole genome sequences and phenotypic features suggested that S. noursei and S. albulus belong to the same species. Therefore, it is proposed that S. albulus is reclassified as a later heterotypic synonym of S. noursei.

Comparative genomics-based probiotic relevance of Limosilactobacillus fermentum KUB-D18.

Limosilactobacillus fermentum KUB-D18 is a heterofermentative lactic acid bacterium that its potential probiotic relevance originally isolated from the chicken intestine. This study sequenced a whole-genome of L. fermentum KUB-D18 and annotated its genes and functions in relation to probiotic properties. As a result, the genome sequence of L. fermentum KUB-D18 approximately contained 2.02 Mbps with GC content of51.7%. After annotating the genome by integrated protein and pathway databases, 2,158 protein-encoding genes were majorly annotated for metabolisms of amino acids, carbohydrates and cofactors as well as vitamins which showed a versatile metabolic capability to gastrointestinal microhabitats. According to the comparative genome analysis of L. fermentum KUB-D18 and the other related strains, L. fermentum KUB-D18 showed common characteristics e.g., folate biosynthesis and bile salt hydrolase enzymes-related cholesterol lowering effect as well as a unique gene cluster involved in metabolism of l-ascorbic acid of L. fermentum KUB-D18. Taken together, L. fermentum KUB-D18 genome provides the genetic basis towards cellular capability for further elucidating the functional mechanisms of its probiotic properties. This study serves for designing desirable targets for the development of probiotic foods and feeds.

Pseudonocardia terrae sp. nov., an actinobacterium isolated from rice rhizosphere soil in Thailand.

A novel actinomycete, designated strain RS11V-5T, was isolated from rhizosphere soil of Oryza sativa L. collected from Roi Et Province, Thailand, and its taxonomic position was evaluated. Cells of strain RS11V-5T were Gram-stain-positive, aerobic, and non-motile. Whole-cell hydrolysates contained meso-diaminopimelic acid, arabinose, galactose, glucose and ribose. MK-8(H4) was detected as the predominant menaquinone of this strain. The polar lipids were diphosphatidylglycerol, phosphatidylmethylethanolamine, phosphatidylethanolamine, hydroxy-phosphatidylmethylethanolamine, hydroxy-phosphatidylethanolamine, an unidentified phospholipid, an unidentified aminolipid and an unidentified glycolipid. The major fatty acids were iso-C16 : 0, C16 : 0 and C16 : 1 ω7c/C16 : 1 ω6c. Phylogenetic analyses based on the 16S rRNA gene sequences showed that strain RS11V-5T belonged to the genus Pseudonocardia and had high 16S rRNA sequence similarity of 99.3 % to Pseudonocardia kujensis KCTC 29062T and less than 98.4 % to other members of the genus Pseudonocardia. The DNA G+C content of the strain RS11V-5T was 73.3 mol%. Strain RS11V-5T showed 46.5 % digital DNA-DNA hybridization, 92.2 % orthologous average nucleotide identity (OrthoANI), 90.2 % ANI based on blast and 92.7 % ANI based on MUMmer to P. kujensis KCTC 29062T. Based its phenotypic, genotypic, phylogenetic and chemotaxonomic characteristics, strain RS11V-5T represents a novel species of the genus Pseudonocardia, for which the name Pseudonocardia terrae sp. nov. is proposed. The type strain is RS11V-5T (=TBRC 15286T=NBRC 115296T).

Acetobacter garciniae sp. nov., an acetic acid bacterium isolated from fermented mangosteen peel in Thailand.

Two isolates, MS16-SU-2T and MS18-SU-3, obtained from fermented mangosteen peel in vinegar were suggested to constitute a new species assignable to the genus Acetobacter based on the results of 16S rRNA gene sequencing. The two isolates showed the highest sequence similarity (98.58%) to Acetobacter tropicalis NBRC 16470T and Acetobacter senegalensis LMG 23690T. However, the calculated similarity values were lower than the threshold for species demarcation. The phylogenetic analysis showed that the branches of the two isolates were separated from other Acetobacter species, and the two isolates constituted a new species in the genus Acetobacter. The genomic DNA of isolate MS16-SU-2T was sequenced. The assembled genome of the isolate was analysed, and the results showed that the highest average nucleotide identity value of 75.9 % was with Acetobacter papayae JCM 25143T and the highest digital DNA-DNA hybridization value of 25.1 % was with Acetobacter fallax LMG 1636T, which were lower than the cutoff values for species delineation. The phylogenetic tree based on the genome sequences showed that the lineage of isolate MS16-SU-2T was most closely related to A. papayae JCM 25143T and Acetobacter suratthaniensis TBRC 1719T, but separated from the branches of these two species. In addition, the two isolates could be distinguished from the type strains of closely related species by their phenotypic characteristics and MALDI-TOF profiles. Therefore, the two isolates, MS16-SU-2T (=TBRC 12339T=LMG 32243T) and MS18-SU-3 (=TBRC 12305), can be assigned to an independent species within the genus Acetobacter, and the name of Acetobacter garciniae sp. nov. is proposed for the two isolates.

Microbispora oryzae sp. nov., isolated from leaves of rice plant (Oryza sativa L.).

An endophytic actinomycete, designated strain RL4-1ST, was isolated from surface-sterilized leaves of rice plant (Oryza sativa L.) collected from Buri Rum province, Thailand. Its taxonomic status was determined using a polyphasic approach. Phylogenetic analysis based on the 16S rRNA gene sequences showed that the strain RL4-1ST belongs to the genus Microbispora and is most closely related to Microbispora rosea subsp. rosea ATCC 12950T (98.5%). The strain forms pairs of spores on short sporophores borne on the aerial mycelium. Polar lipid profile of strain RL4-1ST is diphosphatidylglycerol, hydroxy-phosphatidylethanolamine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylinositol mannosides, ninhydrin-positive glycophospholipid, two unidentified phospholipids, an unidentified aminolipid, and an unidentified glycolipid. MK-9(H4), MK-9(H2), and MK-9 are major menaquinones of this organism. The predominant cellular fatty acids are iso-C16:0, C17:0 and C16:0. Strain RL4-1ST contains meso-diaminopimelic acid, glucose, madurose and ribose in whole-cell hydrolysates. The draft genome of strain RL4-1ST consists of 7.46 Mbp and has a G + C content 71.2 mol%. Digital DNA-DNA hybridization and average nucleotide identity values between the genome sequence of strain RL4-1ST with Microbispora rosea subsp. rosea ATCC 12950T are 26.0% and 80.7%, respectively. Based on data of genotypic, phenotypic, phylogenetic and chemotaxonomic analysis, strain RL4-1ST represents a novel species of the genus Microbispora, for which the name Microbispora oryzae sp. nov. is proposed. The type strain is RL4-1ST (= TBRC 14817T = NBRC 115115T).

Gordonia oryzae sp. nov., isolated from rice plant stems (Oryza sativa L.).

A novel endophytic actinomycete, designated strain RS15-1ST, was isolated from surface-sterilized stems of Oryza sativa L. collected from Sisaket province, Thailand. The colony of strain was strong orange, catalase-positive and oxidase-negative. Growth occurred at a temperature range of 17-37 °C, at pH 4.0-9.0 and in the presence of 0-13 % (w/v) NaCl. Phylogenetic analyses based on the 16S rRNA sequences showed that strain RS15-1ST belonged to the genus Gordonia and was closely related to Gordonia polyisoprenivorans DSM 44302T (98.8 %) and Gordonia rhizosphera DSM 44383T (98.4 %). The major cellular fatty acids were C16 : 0, C18 : 0 10-methyl (tbsa), C16 : 1ω7c/C16 : 1ω6c and C18 : 1ω9c. The menaquinones were MK-9(H2) and MK-8(H2). Strain RS15-1ST contained meso-diaminopimelic acid, arabinose, galactose, mannose and ribose in whole-cell hydrolysates. The polar lipids of the strain were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol mannosides, an unidentified polar lipid and two unidentified phospholipids. The DNA G+C content was 66.3 mol%. In silico DNA-DNA hybridization of strain RS15-1ST showed 48.3 and 20.5 % relatedness to its closest neighbours, Gordonia polyisoprenivorans DSM 44302T and Gordonia rhizosphera DSM 44383T, respectively. Based on data of genotypic, phenotypic, phylogenetic and chemotaxonomic analysis, strain RS15-1ST represents a novel species of the genus Gordonia, for which the name Gordonia oryzae sp. nov. is proposed. The type strain is RS15-1ST (=TBRC 8485T=NBRC 113446T).

Quadrisphaera oryzae sp. nov., an endophytic actinomycete isolated from leaves of rice plant (Oryza sativa L.).

A novel endophytic actinobacterium, designated strain RL12-1ST, was isolated from surface-sterilized leaves of Oryza sativa L. collected from Roi Et province, Thailand. Taxonomic position of strain RL12-1ST was studied using the polyphasic approach. Phylogenetic analyses base on the 16S rRNA gene sequences showed that strain RL12-1ST belonged to the genus Quadrisphaera and was closely related to Quadrisphaera granulorum AG019T (99.0%). The colony of strain RL12-1ST was vivid orange. Growth occurred at a temperature between 17-37 °C and pH between 6.0-8.0. The strain contained meso-diaminopimelic acid and galactose, glucose, mannose, ribose and rhamnose in whole-cell hydrolysates. The predominant menaquinone was MK-8(H2). The major fatty acids were C16:1ω7c/C16:1ω6c, C16:0 and C18:1ω7c. The polar lipids of the strain were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol and phosphatidylinositol mannosides. The DNA G + C content of genomic DNA was 76.1 mol%. Based on comparative analysis of physiological, biochemical and chemotaxonomic data, including DNA-DNA hybridization, strain RL12-1ST represents a novel species of the genus Quadrisphaera, for which the name Quadrisphaera oryzae sp. nov. is proposed. The type strain is RL12-1ST ( = TBRC 8486T = NBRC 113447T).

Nocardiopsissediminis sp. nov., isolated from mangrove sediment.

A filamentous actinomycete, designated strain 1SS5-02T, was isolated from mangrove sediment collected from Ranong province, Thailand. The strain formed aerial and substrate mycelia composed of long, branched hyphae. Aerial mycelia differentiated into non-motile, rod-shaped spores. The organism contained meso-diaminopimelic acid and no diagnostic sugars in whole-cell hydrolysates. The predominant menaquinones were MK-11(H4), MK-11(H6) and MK-11(H8). Polar lipids comprised phosphatidylcholine, phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, two unidentified phospholipids and four unidentified lipids. The major fatty acids were iso-C16 : 0, C18 : 1ω9c, 10-methyl C18 : 0 and anteiso-C17 : 0. The G+C content of the genomic DNA was 73.5 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain 1SS5-02T belonged to the genus Nocardiopsis. The strain showed the highest degree of 16S rRNA gene sequence similarity with 'Nocardiopsis mangrovei' HA11166 (97.9 %) and Nocardiopsis trehalosi VKM Ac-942T (97.8 %). However, strain 1SS5-02T could be distinguished from its nearest phylogenetic relatives in the genus Nocardiopsis on the basis of DNA-DNA relatedness values and the combination of phenotypic properties. On the basis of polyphasic taxonomy, strain 1SS5-02T is considered to represent a novel species of the genus Nocardiopsis, for which the name Nocardiopsis sediminis sp. nov. is proposed. The type strain is 1SS5-02T (=BCC 75410T=NBRC 110934T).

Kineococcus mangrovi sp. nov., isolated from mangrove sediment.

An aerobic, Gram-stain-positive, motile, coccus-shaped actinobacterium, designated strain L2-1-L1T, was isolated from mangrove sediment in Thailand. The organism was deep orange, oxidase-negative and catalase-positive. Growth occurred at temperatures between 17 and 32 °C and with NaCl concentrations up to 10 %. Chemotaxonomic characteristics of strain L2-1-L1T were typical of the genus Kineococcus. The diagnostic diamino acid of the peptidoglycan was meso-diaminopimelic acid. Whole-cell hydrolysates contained arabinose, galactose, glucose, mannose and ribose. The menaquinone was MK-9(H2). Mycolic acids were not detected. Anteiso-C15 : 0 and iso-C14 : 0 were detected as the major cellular fatty acids. The major polar lipids comprised diphosphatidylglycerol, phosphatidylglycerol and an unidentified phosphoglycolipid. The G+C content of the DNA was 74.7 mol%. A phylogenetic tree, based on 16S rRNA gene sequences, revealed that strain L2-1-L1T represents a novel member of the genus Kineococcus. The most closely related species were Kineococcus endophytica KLBMP 1274T, Kineococcus aurantiacus NBRC 15268T and Kineococcus rhizosphaerae RP-B16T, with 98.9, 98.6 and 98.5 % 16S rRNA gene sequence similarity, respectively. On the basis of phylogenetic analysis, DNA-DNA relatedness data, phenotypic characteristics and chemotaxonomic data, a novel species of the genus Kineococcus is proposed: Kineococcus magrovi sp. nov. The type strain is strain L2-1-L1T ( = BCC 75409T = NBRC 110933T).

Melanogenic actinomycetes from rhizosphere soil-antagonistic activity against Xanthomonas oryzae and plant-growth-promoting traits.

A total of 210 melanogenic actinomycetes were isolated from 75 rhizospheric soils using ISP6 and ISP7 agar supplemented with antifungal and antibacterial agents. Their morphological characteristics and the presence of ll-diaminopimelic acid in whole-cell hydrolyzates revealed that all isolates belonged to the genus Streptomyces. Their ability to inhibit the growth of 2 pathogenic rice bacteria, Xanthomonas oryzae pv. oryzae and Xanthomonas oryzae pv. oryzicola, was observed using the agar overlay method. The results indicated that 61.9% of the isolates could inhibit at least one of the tested rice pathogens. Among these, isolate TY68-3 showed the highest antibacterial activity and siderophore production. The 16S rRNA gene sequence analysis of 46 representative isolates revealed that isolates with high similarity to Streptomyces bungoensis were frequently found. The present study indicated the potential of melanogenic actinomycetes for use as biocontrol agents against X. oryzae as well as their diversity in rhizospheric soils.