DNA-DNA Hybridization Data Research Articles

Aurantiacibacter flavus sp. nov. and Aurantiacibacter gilvus sp. nov., isolated from the mudflat of Suaeda japonica colonies.

Two novel strains were isolated from the mudflat of Suaeda japonica colonies in Incheon, Republic of Korea. Designated as DGU5T and DGU6T, these strains were Gram-stain-negative, facultatively anaerobic and rod-shaped and had yellowish colonies. Both strains were determined to belong to the genus Aurantiacibacter through phylogenetic analysis of their 16S rRNA sequences and draft genomes. The cells of strain DGU5T were non-motile and grew at temperatures ranging between 7-45°C (optimum, 25-30°C), pH 6.0-10.0 (optimum, 7.0-8.0) and in the presence of 0-11.0% NaCl (optimum, 2.0%). The cells of strain DGU6T were non-motile and grew in temperatures ranging from 10-45 °C (optimum, 30-35°C), pH 3.0-10.0 (optimum, 7.0-8.0) and in the presence of 0-11.0% NaCl (optimum, 2.0%). Overall genome relatedness index calculations revealed average nucleotide identity values (72.3-88.6%) and digital DNA-DNA hybridization values (18.8-35.9%) aligning with those of the genus Aurantiacibacter. The major fatty acids in both strains were C17:1 ω6c and summed feature 8 (C18:1 ω6c/C18:1 ω7c), while the predominant polar lipids were sphingoglycolipid, phosphatidylglycerol, and diphosphatidylglycerol. Phylogenetic, average nucleotide identity, digital DNA-DNA hybridization, physiological, and biochemical data collectively demonstrated the distinctiveness of the novel strains from other members within the family Erythrobacteraceae. We propose the names A. flavus sp. nov. (type strain DGU5T = KACC 23720T = TBRC 19015T) and A. gilvus sp. nov. (type strain DGU6T = KACC 23721T = TBRC 19016T) for the two strains.

Cellulomonas alba sp. nov. and Cellulomonas edaphi sp. nov., isolated from wetland soils.

Two novel strains were isolated from wetland soils in Goyang, Republic of Korea. The two Gram-stain-positive, facultatively anaerobic, rod-shaped bacterial-type strains were designated MW4T and MW9T. Phylogenomic analysis based on whole-genome sequences suggested that both strains belonged to the genus Cellulomonas. The cells of strain MW4T were non-motile and grew at 20-40 °C (optimum, 35 °C), at pH 6.0-10.0 (optimum, pH 8.0) and in the presence of 0-1.0% NaCl (optimum, 0 %). The cells of strain MW9T were non-motile and grew at 20-40 °C (optimum, 35 °C), at pH 5.0-9.0 (optimum, pH 8.0) and in the presence of 0-1.0% NaCl (optimum, 0 %). The average nucleotide identity (77.1-88.1 %) and digital DNA-DNA hybridization values (21.0-34.8 %) between the two novel strains and with their closely related strains fell within the range for the genus Cellulomonas. The novel strains MW4T and MW9T and reference strains possessed alkane synthesis gene clusters (oleA, oleB, oleC and oleD). Phylogenomic, phylogenetic, average nucleotide identity, digital DNA-DNA hybridization, physiological and biochemical data indicated that the novel strains were distinct from other members of the family Cellulomonadaceae. We propose the names Cellulomonas alba sp. nov. (type strain MW4T=KACC 23260T=TBRC 17645T) and Cellulomons edaphi sp. nov. (type strain MW9T=KACC 23261T=TBRC 17646T) for the two strains.

Sphingobium agri sp. nov., isolated from rhizospheric soil of eggplant.

A Gram-stain-negative, aerobic, short rod-shaped and motile bacterial strain, designated MAH-33T, was isolated from rhizospheric soil of eggplant. The colonies were observed to be yellow-coloured, smooth, spherical and 0.1-0.3 mm in diameter when grown on TSA agar medium for 2 days. Strain MAH-33T was found to be able to grow at 10-40 °C, at pH 5.0-10.0 and at 0-3.0 % NaCl (w/v). The strain was found to be positive for both oxidase and catalase tests. The strain was positive for hydrolysis of tyrosine and aesculin. According to the 16S rRNA gene sequence comparisons, the isolate was identified as a member of the genus Sphingobium and to be closely related to Sphingobium quisquiliarum P25T (98.4 % similarity), Sphingobium mellinum WI4T (97.8 %), Sphingobium fuliginis TKPT (97.3 %) and Sphingobium herbicidovorans NBRC 16415T (96.9 %). The novel strain MAH-33T has a draft genome size of 3 908 768 bp (28 contigs), annotated with 3689 protein-coding genes, 45 tRNA and three rRNA genes. The average nucleotide identity and digital DNA-DNA hybridization values between strain MAH-33T and closely related type strains were in the range of 79.8-81.6 % and 23.2-24.5 %, respectively. The genomic DNA G+C content was determined to be 62.2 %. The predominant isoprenoid quinone was ubiquinone 10. The major fatty acids were identified as C16 : 0 and summed feature 8 (C18 : 1 ω7c and/or C18 : 1 ω6c). The polar lipids identified in strain MAH-33T were phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, sphingoglycolipid, phosphatidylcholine; one unknown phospholipid and one unknown lipid. On the basis of digital DNA-DNA hybridization, ANI value, genotypic analysis, chemotaxonomic and physiological data, strain MAH-33T represents a novel species within the genus Sphingobium, for which the name Sphingobium agri sp. nov. is proposed, with MAH-33T (=KACC 19973T = CGMCC 1.16609T) as the type strain.

Genome insights into the pharmaceutical and plant growth promoting features of the novel species Nocardia alni sp. nov

BackgroundRecent studies highlighted the biosynthetic potential of nocardiae to produce diverse novel natural products comparable to that of Streptomyces, thereby making them an attractive source of new drug leads. Many of the 119 Nocardia validly named species were isolated from natural habitats but little is known about the diversity and the potential of the endophytic nocardiae of root nodule of actinorhizal plants.ResultsThe taxonomic status of an actinobacterium strain, designated ncl2T, was established in a genome-based polyphasic study. The strain was Gram-stain-positive, produced substrate and aerial hyphae that fragmented into coccoid and rod-like elements and showed chemotaxonomic properties that were also typical of the genus Nocardia. It formed a distinct branch in the Nocardia 16S rRNA gene tree and was most closely related to the type strains of Nocardia nova (98.6%), Nocardia jiangxiensis (98.4%), Nocardia miyuensis (97.8%) and Nocardia vaccinii (97.7%). A comparison of the draft genome sequence generated for the isolate with the whole genome sequences of its closest phylogenetic neighbours showed that it was most closely related to the N. jiangxiensis, N. miyuensis and N. vaccinii strains, a result underpinned by average nucleotide identity and digital DNA-DNA hybridization data. Corresponding taxogenomic data, including those from a pan-genome sequence analysis showed that strain ncl2T was most closely related to N. vaccinii DSM 43285T. A combination of genomic, genotypic and phenotypic data distinguished these strains from one another. Consequently, it is proposed that strain ncl2T (= DSM 110931T = CECT 30122T) represents a new species within the genus Nocardia, namely Nocardia alni sp. nov. The genomes of the N. alni and N. vaccinii strains contained 36 and 29 natural product-biosynthetic gene clusters, respectively, many of which were predicted to encode for a broad range of novel specialised products, notably antibiotics. Genome mining of the N. alni strain and the type strains of its closest phylogenetic neighbours revealed the presence of genes associated with direct and indirect mechanisms that promote plant growth. The core genomes of these strains mainly consisted of genes involved in amino acid transport and metabolism, energy production and conversion and transcription.ConclusionsOur genome-based taxonomic study showed that isolate ncl2T formed a new centre of evolutionary variation within the genus Nocardia. This novel endophytic strain contained natural product biosynthetic gene clusters predicted to synthesize novel specialised products, notably antibiotics and genes associated with the expression of plant growth promoting compounds.

Comparative Genome Analysis of the Genus Thiothrix Involving Three Novel Species, Thiothrix subterranea sp. nov. Ku-5, Thiothrix litoralis sp. nov. AS and "Candidatus Thiothrix anitrata" sp. nov. A52, Revealed the Conservation of the Pathways of Dissimilatory Sulfur Metabolism and Variations in the Genetic Inventory for Nitrogen Metabolism and Autotrophic Carbon Fixation.

Two strains of filamentous, colorless sulfur bacteria were isolated from bacterial fouling in the outflow of hydrogen sulfide-containing waters from a coal mine (Thiothrix sp. Ku-5) and on the seashore of the White Sea (Thiothrix sp. AS). Metagenome-assembled genome (MAG) A52 was obtained from a sulfidic spring in the Volgograd region, Russia. Phylogenetic analysis based on the 16S rRNA gene sequences showed that all genomes represented the genus Thiothrix. Based on their average nucleotide identity and digital DNA-DNA hybridization data these new isolates and the MAG represent three species within the genus Thiothrix with the proposed names Thiothrix subterranea sp. nov. Ku-5T, Thiothrix litoralis sp. nov. AST, and “Candidatus Thiothrix anitrata” sp. nov. A52. The complete genome sequences of Thiothrix fructosivorans QT and Thiothrix unzii A1T were determined. Complete genomes of seven Thiothrix isolates, as well as two MAGs, were used for pangenome analysis. The Thiothrix core genome consisted of 1,355 genes, including ones for the glycolysis, the tricarboxylic acid cycle, the aerobic respiratory chain, and the Calvin cycle of carbon fixation. Genes for dissimilatory oxidation of reduced sulfur compounds, namely the branched SOX system (SoxAXBYZ), direct (soeABC) and indirect (aprAB, sat) pathways of sulfite oxidation, sulfur oxidation complex Dsr (dsrABEFHCEMKLJONR), sulfide oxidation systems SQR (sqrA, sqrF), and FCSD (fccAB) were found in the core genome. Genomes differ in the set of genes for dissimilatory reduction of nitrogen compounds, nitrogen fixation, and the presence of various types of RuBisCO.

Micromonospora veneta sp. nov., an endophytic actinobacterium with potential for nitrogen fixation and for bioremediation

Strain CAP181T, an endophytic actinobacterium, was isolated from a surface sterilized root sample of a native pine tree, Flinders University, Adelaide, South Australia. Chemotaxonomic data including cell wall components, major fatty acids, and major menaquinones confirmed the affiliation of strain CAP181T to the genus Micromonospora. This strain was Gram stain positive with well-developed substrate mycelia to forma single spore with hairy surface. The phylogenetic tree showed that M. coerulea NBRC 13504T is the closest phylogenetic neighbour, sharing 99.2% 16S rRNA gene similarity and the next closest neighbor is M. chaiyaphumensis DSM 45246T (98.7%). Genome mining of this strain revealed genes encoding to enzymes relating to nitrogen fixation and bioremediation. Based on genotypic and phenotypic studies including DNA-DNA hybridization data, strain CAP181T was different from any of the closely related species with valid names. The name proposed for the new species is Micromonospora veneta sp. nov. The type strain is CAP181T (= DSM 109713T = NRRL B-65535T).

Fusobacterium watanabei sp. nov. As additional species within the genus Fusobacerium, isolated from human clinical specimens

Eight spindle-shaped bacteria were isolated from clinical samples in Japan and investigated for their taxonomic position. Phylogenetic trees (based on 16S rRNA, rpoB, zinc protease, and gyrB gene sequence comparisons) showed distinct clustering of eight strains with the type strain of Fusobacterium nucleatum and its closely related species. In silico whole genome comparison analysis based on average nucleotide index based on BLAST (ANIb) and digital DNA-DNA hybridization (dDDH) data between our clinical isolates (PAGU 1795, PAGU 1796T, and PAGU 1797) and the type strain of the closely related species showed values of less than 92.4% and 49.5%, respectively. On the basis of its phylogenetic and genomic distinctiveness together with differential phenotypic properties and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS) characteristic signal patterns, we propose Fusobacterium watanabei sp. nov., with the type strain PAGU 1796T (= GTC 21791T = CCUG 74246T).

Taxogenomics and Systematics of the Genus Pantoea.

Members of the genus Pantoea are Gram-negative bacteria isolated from various environments. Taxonomic affiliation based on multilocus sequence analysis (MLSA) is used routinely for inferring accurate phylogeny and identification of bacterial species and genera. Partial sequences of five housekeeping genes (fusA, gyrB, leuS, rpoB, and pyrG) were extracted from 206 draft or complete genomes of Pantoea strains publicly available in databases and analyzed together with the representative sequences of the 25 validly published Pantoea type strains to verify and assess their phylogenetic assignations. Of a total of 159 strains assigned to species level, 11.3% of the non-type strains were incorrectly assigned within suitable Pantoea species. The highest proportion of misidentified strains was recorded in Pantoea vagans, 8 out of 15 (53.3%) inaccurate assignations at the species level. One probable reason for this incorrect classification could be the method previously used for strain identification. Forty-seven (22.8%) genome sequences were from strains identified at the genus level only (Pantoea sp.). A combination of MLSA, average nucleotide identities [ANI and MuMmer-based ANI (ANIm)], tetranucleotide usage pattern (TETRA), and genome-based DNA-DNA hybridization (gDDH) data was used to accurately assign 25 of the 47 strains to validly published Pantoea species, while 17 strains could be assigned as putative novel species within the genus Pantoea. Four genomes designed as Pantoea sp. were identified as Mixta calida. Positive and significant correlation coefficients were computed between MLSA and all the indices derived from whole-genome sequences being proposed for species delimitation. gDDH exhibited the best correlation with MLSA while TETRA was the worst. Accurate species-level identification is key to a better understanding of bacterial diversity and evolution. The MLSA scheme used here could be instrumental to determine the correct taxonomic status of new whole-genome sequenced Pantoea strains, especially non-type strains, before depositing into public databases.

Thermoflavimicrobium daqui sp. nov., a thermophilic microbe isolated from Moutai-flavour Daqu.

Moutai-flavour Daqu is an important starter to support growth of microorganisms in the fermented process of Moutai-flavour liquor. A novel thermophilic microorganism, designated strain FBKL4.011T, was isolated from Moutai-flavour Daqu samples collected from Guotai distillery in Renhuai, Guizhou province, south-west China. The strain could grow at 45-65 °C (optimum 45 °C). Based on polyphasic analysis, strain FBKL4.011T was affiliated to the genus Thermoflavimicrobium. It formed abundant pale-yellow aerial and substrate mycelium, bearing single endospores (7.0-10.0 µm diameter) on branched long sporophores (5.0 µm diameter). The cell-wall peptidoglycan contained meso-diaminopimelic acid; ribose, glucose and mannose were the primary whole-cell sugars. The major fatty acids were iso-C14 : 0, iso-C15 : 0, anteiso-C15 : 0 and iso-C16 : 0. The predominant menaquinone were MK-8 and MK-9. The polar phospholipids contained diphosphatidyl glycerol, phosphatidyl ethanolamine, one unidentified phospholipid and one unidentified lipid. The G+C content of the genome was 43.1 mol%. According to the 16S rRNA gene sequence analysis, strain FBKL4.011T was closely related to Thermoflavimicrobium dichotomicum JCM 9688T (95.3 % sequence similarity), and other members within the family Thermoactinomycetaceae (less than 93.0 % sequence similarity). The DNA-DNA hybridisation data showed low relatedness between strain FBKL4.011T and T. dichotomicum JCM 9688T, Laceyella sacchari KCTC 9790T, Laceyella tengchongensis YIM 10002T, Laceyella sediminis RHA1T(36.7±1.1 %, 30.0±1.2 %, 21.3±2.1 % % and 37.6±0.9 %, respectively). Based on data from the polyphasic analysis, strain FBKL4.011T is considered to represent a novel species of the genus Thermoflavimicrobium, for which the name Thermoflavimicrobium daqui sp. nov. is proposed. The type strain is FBKL4.011T(=KCTC 43036T=CICC 24504T).

Erythrobacter suaedae sp. nov., isolated from a rhizosphere mudflat of a halophyte (Suaeda japonica).

The taxonomic status of a Gram-reaction-negative, aerobic, motile, rod-shaped bacterium, designated strain GH3-15T, was examined by a polyphasic approach. The strain, which was isolated from the rhizosphere mudflat of a halophyte at the seashore of Gangwha Island, Republic of Korea, was found to belong to the family Erythrobacteraceae based on 16S rRNA gene sequences. The closest phylogenetic neighbour was Erythrobacter xanthus SM1501T (98.3 % sequence similarity). Levels of 16S rRNA gene sequence similarity of strain GH3-15T to other members of the family Erythrobacteracae were <97.1 %. The respiratory quinone was Q-10. The major fatty acids were C17 : 1ω6c, C18 : 1ω7c, C15 : 0 2-OH, 11-methyl C18 : 1ω7c and C17 : 0. The polar lipids were phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol and sphingoglycolipid. The novel isolate exhibited growth at 20-40 °C, at pH 5-9, and in the presence of 1-7 % (w/v) NaCl. DNA relatedness between strain GH3-15T and its closet relative was 32.9±8.8 %. On the basis of phenotypic, chemotaxonomic and DNA-DNA hybridization data, in addition to a distinct phylogenetic position, strain GH3-15T (=KCTC 62380T=JCM 32445T) represents a novel species of the genus Erythrobacter, for which the name Erythrobactersuaedae sp. nov. is proposed.

Genetic analysis of Ximenia americana based on the cross-species DNA-DNA hybridization technique

Abstract. Ekandjo A, Naomand E, Kahaka G. 2018. Genetic analysis of Ximenia americana based on DNA-DNA based on the cross-species hybridization technique. Cell Biol Dev 2: 1-7. Genetic analysis of species without the availability of complete genome arrays can be studied using the cross-species hybridization technique. The preliminary microarray studies have created DNA-DNA hybridization of X. americana to the GeneChip of Arabidopsis thaliana (ATH1). Based on the perfect-match (PM) probe signal, the probe pairs of A. thaliana that hybridized with the genomic DNA of X. americana were selected and analyzed using a cel file parser script. This selection was to generate new high-density probe mask files. The X. americana DNA-DNA hybridization data were effectively represented by this file. In addition, this study carried out gene ontology analysis of hybridization data of X. americana/A. thaliana. This gene ontology analysis showed that genes for abiotic stress response were over-expressed in respective comparison to model species of A. thaliana under natural conditions. This analysis was confirmed independently with PCR amplification of the orthologous genes using genomic DNA of X. americana, such as AT4G15910.1, SAD2, HXK1, ACC, and ERF/AP2. Due to the lack of genomic sequence data in X. americana, the primers for genomic amplification were designed using a genomic sequence of A. thaliana. The primers were designed to produce a genomic PCR product of 100 bp. Each selected gene was successfully amplified. Therefore, it gave evidence of homology within primer binding sites. However, the genomic amplification of these crucial abiotic factors in X. americana confirms the response type, which supports the adaptation of X. americana under natural conditions with stress associated (with heat and drought stress). Although transcript levels of these unique abiotic response factors could not be observed in absolute or relative terms, the study showed the inherent existence of such genes at the genomic level of X. americana. Further studies are needed to confirm that some modification of the factors of these abiotic responses or other abiotic responses within the genome of X. americana gives the key basis for its adaptation to the comparatively dry and hot climatic conditions.

Lysobacter tabacisoli sp. nov., isolated from rhizosphere soil of Nicotiana tabacum L.

A Gram-stain-negative, aerobic, rod-shaped bacterium, designated strain C8-1T, was isolated from the rhizosphere soil of Nicotiana tabacum L. collected from Kunming, south-west China. The cells showed oxidase-positive and catalase-positive reactions. Growth was observed at 10-40 °C, at pH 6.0-8.0 and in the presence of up to 1 % (w/v) NaCl, with optimal growth at 30 °C and pH 7.0. The predominant isoprenoid quinone was Q-8. The major fatty acids were identified as iso-C15 : 0, iso-C17 : 0 and iso-C17 : 1ω9c. The cellular polar lipids contained diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylmethylethanolamine, five unidentified phospholipids and two unidentified aminophospholipids. The genomic DNA G+C content was 70.7 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain C8-1T should be assigned to the genus Lysobacter. 16S rRNA gene sequence similarity analysis showed that strain C8-1T was closely related to Lysobacter cavernae YIM C01544T (98.6 %), Lysobacter soli DCY21T (97.6 %), Lysobacter panacisoli CJ29T (97.3 %), Lysobacter firmicutimachus PB-6250T (97.3 %), Lysobacter niastensis GH41-7T (97.3 %) and Lysobacter gummosus KCTC 12132T (97.1 %). DNA-DNA hybridization data indicated that the isolate may represent a novel genomic species belonging to the genus Lysobacter. Polyphasic taxonomic characteristics indicated that strain C8-1T represents a novel species of the genus Lysobacter, for which the name Lysobacter tabacisoli sp. nov. is proposed. The type strain is C8-1T (=KCTC 62034T=CGMCC 1.16271T) .

Bhargavaea changchunensis sp. nov. isolated from soil in China.

A Gram-staining-positive, aerobic, rod-shaped (201802YP6T) bacteria was isolated from soil, Northeast of China. Growth occurred at 10-40°C (optimum 25-30°C), at pH 6.0-8.0 (optimum 7.0) and at 0-2% NaCl. Based on 16S rRNA gene sequence analysis, the nearest phylogenetic neighbors of strain 201802YP6T were identified as Bhargavaea cecembensis DSE10T (99.52%), Bhargavaea beijingensis ge10T (99.45%), Bhargavaea indica KJW98T (99.45%), Bhargavaea ullalensis ZMA19T (98.81%), and Bhargavaea ginsengi ge14T (98.76%). Levels of similarity among strain 201802YP6T and other Bhargavaea species were lower than 98.76%. GyrB amino acid sequence-based analysis supported the phylogenetic position and also distinguished strain 201802YP6T from the other species of the genus Bhargavaea. DNA-DNA hybridization values between strain 201802YP6T and B. cecembensis, B. beijingensis, B. indica, B. ullalensis, B. ginsengi were 43.5%, 43%, 32.5%, 30.5% and 20.4%, respectively. The DNA G + C content of strain 201802YP6T was 51.23mol%. The average nucleotide identity (ANI) of the draft genome was 87.04% to B. cecembensis DSE10T. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, glycolipids, phosphatidylethanolamine, and phosphatidyllipid. The predominant menaquinone was MK-8. The major fatty acids were iso-C15:0 (39.91%), anteiso-C15:0 (28.86%), anteiso-C17:0 (6.30%) and C16:0 (6.13%). On the basis of the phylogenetic analysis, chemotaxonomic data, physiological characteristics and DNA-DNA hybridization data, strain 201802YP6T represents a novel species of the genus Bhargavaea, for which the name Bhargavaea changchunensis sp. nov. is proposed. The type strain is 201802YP6T (= CGMCC 1.13508T = KCTC 33975T).

Aestuariibaculum marinum sp. nov., a marine bacterium isolated from seawater in South Korea.

A Gram-negative, non-motile, aerobic bacterium, designated strain IP7T, was isolated from seawater at the shore of the Incheon Eulwang-ri beach, South Korea. Cells of strain IP7T are straight or slightly rod-shaped and colonies are round, convex and orange-yellow. Strain IP7T is flexirubin-negative, mild halophile, catalase-and oxidase-positive, and produces a yellow-orange carotenoid pigment. Growth is optimal at 30°C, pH 7-9, and 2.0-4.0% NaCl (w/v). On the basis of 16S rRNA gene sequence similarity, strain IP7T is affiliated with genus Aestuariibaculum in the family Flavobacteriaceae, the closest relative being Aestuariibaculum suncheonense SC17T (98.3% sequence similarity). The DNA G + C content of the novel strain is 37.4 mol%. The only quinone is MK-6 menaquinone. Iso-branched C15:0, iso-branched C15:1 G, and iso-branched C17:0 3-OH are major fatty acids. The major polar lipids are phosphatidylethanolamine, an unidentified aminoglycolipid and two unidentified glycolipids. The DNA-DNA hybridization value of strain IP7T with Aestuariibaculum suncheonense SC17T is 28.87%. Based on the collective DNA-DNA hybridization, biochemical, phylogenetic and physiological data, we report a novel species of the genus Aestuariibaculum for which the name Aestuariibaculum marinum sp. nov. is proposed. The type strain is IP7T (= KCTC 52521T = JCM 31725T).

Halomonas tibetensis sp. nov., isolated from saline lakes on Tibetan Plateau.

Strains pyc13T and ZGT13 were isolated from Lake Pengyan and Lake Zigetang on Tibetan Plateau, respectively. Both strains were Gram-negative, catalase- and oxidase-positive, aerobic, rod-shaped, nonmotile, and nonflagellated bacteria. Phylogenetic analysis based on 16S rRNA gene sequences showed that strains pyc13T and ZGT13 belong to the genus Halomonas, with Halomonas alkalicola 56-L4-10aEnT as their closest neighbor, showing 97.4% 16S rRNA gene sequence similarity. The predominant respiratory quinone of both strains was Q-9, with Q-8 as a minor component. The major fatty acids of both strains were C18:1ω6c/C18:1ω7c, C16:1ω6c/C16:1ω7c, C16:0, and C12:0 3OH. The polar lipids of both strains consisted of phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, glycolipid, phospholipids of unknown structure containing glucosamine, and unidentified phospholipids. The DNA G + C content of pyc13T and ZGT13 were 62.6 and 63.4 mol%, respectively. The DNA-DNA hybridization values of strain pyc13T were 34, 41, 61, 35, and 35% with the reference strains H. alkalicola 56-L4-10aEnT, H. sediminicola CPS11T, H. mongoliensis Z-7009T, H. ventosae Al12T, and H. fontilapidosi 5CRT, respectively. Phenotypic, biochemical, genotypic, and DNA-DNA hybridization data showed that strains pyc13T and ZGT13 represent a new species within the genus Halomonas, for which the name H. tibetensis sp. nov. is proposed. The type strain is pyc13T (= CGMCC 1.15949T = KCTC 52660T).

Scopulibacillus cellulosilyticus sp. nov., a cellulose-degrading bacterium isolated from tea.

A Gram-stain positive, aerobic, non-motile, endospore-forming and rod-shaped strain (THG-NT9T) was isolated from a green tea sample. Growth occurred at 20-45°C (optimum 28-35°C), at pH 6.0-8.0 (optimum 7.0) and at 0-2.0% NaCl (optimum 0%). Based on 16S rRNA gene sequence analysis, the near phylogenetic neighbours of strain THG-NT9T were identified as Scopulibacillus daqui DSM 28236T (98.6%), Scopulibacillus darangshiensis DSM 19377T (97.4%), Pullulanibacillus pueri CGMCC 1.12777T (96.7%) and Pullulanibacillus camelliae CGMCC 1.15371T (96.3%). The DNA G + C content of strain THG-NT9T was determined to be 47.5mol%. DNA-DNA hybridization values between strain THG-NT9T and S. daqui DSM 28236T, S. darangshiensis DSM 19377T, P. pueri CGMCC 1.12777T, P. camelliae CGMCC 1.15371T and Pullulanibacillus naganoensis DSM 10191T were 41.3 ± 0.1 (39.4 ± 0.4% reciprocal analysis), 39.1 ± 0.1 (37.3 ± 0.1%), 21.4 ± 0.7 (20.1 ± 0.3%), 20.7 ± 0.1 (20.1 ± 0.4%) and 12.1 ± 0.2% (8.3 ± 0.2%). The polar lipids were identified as diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and three unidentified lipids. The quinone was identified as MK-7. The major fatty acids were C18:3 ω7c, iso-C15:0, iso-C16:0, iso-C17:0 and anteiso-C17:0. The cell wall type was determined to be A1γ peptidoglycan with meso-diaminopimelic acid as the diagnostic diamino acid plus alanine and glutamic acid and glucose as the cell wall sugar. On the basis of the phylogenetic analysis, chemotaxonomic data, physiological characteristics, and DNA-DNA hybridization data, strain THG-NT9T represents a novel species of the genus Scopulibacillus, for which the name Scopulibacillus cellulosilyticus sp. nov. is proposed. The type strain is THG-NT9T (= KCTC 33918T = CGMCC 1.16305T).

Nibribacter flagellatus sp. nov., isolated from rhizosphere of Hibiscus syriacus and emended description of the genus Nibribacter.

A Gram-stain negative, aerobic, motile by flagella, rod-shaped strain (THG-T16T) was isolated from rhizosphere of Hibiscus syriacus. Growth occurred at 10-40°C (optimum 28-30°C), at pH 6.0-8.0 (optimum 7.0) and at 0-1.0% NaCl (optimum 0%). Based on 16S rRNA gene sequence analysis, the near phylogenetic neighbours of strain THG-T16T were identified as Nibribacter koreensis KACC 16450T (98.6%), Rufibacter roseus KCTC 42217T (94.7%), Rufibacter immobilis CCTCC AB 2013351T (94.5%) and Rufibacter tibetensis CCTCC AB 208084T (94.4%). The DNA G+C content of strain THG-T16T was determined to be 46.7mol%. DNA-DNA hybridization values between strain THG-T16T and N. koreensis KACC 16450T, R. roseus KCTC 42217T, R. immobilis CCTCC AB 2013351T, R.tibetensis CCTCC AB 208084T were 33.5 ± 0.5% (31.7 ± 0.7% reciprocal analysis), 28.1 ± 0.2% (25.2 ± 0.2%), 17.1 ± 0.9% (10.2 ± 0.6%) and 8.1 ± 0.3% (5.2 ± 0.1%). The polar lipids were identified as phosphatidylethanolamine, two unidentified aminophospholipids, an unidentified aminolipid and three unidentified lipids. The quinone was identified as MK-7 and the polyamine as sym-homospermidine. The major fatty acids were identified as C16:1 ω5c, C17:1 ω6c, iso-C15:0, summed feature 3 (C16:1 ω7c and/or C16:1 ω6c) and summed feature 4 (iso-C17:1 I and/or anteiso-C17:1 B). On the basis of the phylogenetic analysis, chemotaxonomic data, physiological characteristics, and DNA-DNA hybridization data, strain THG-T16T represents a novel species of the genus Nibribacter, for which the name Nibribacter flagellatus sp. nov. is proposed. The type strain is THG-T16T(= KACC 19188T = CCTCC AB 2016246T).

Paracoccus pueri sp. nov., isolated from Pu'er tea.

A Gram-stain negative, aerobic, short rod-shaped, motile by flagella bacterial strain (THG-N2.35T), was isolated from Pu'er tea. Growth occurred at 10-40°C (optimum 28°C), at pH 4-7 (optimum 7) and at 0-5% NaCl (optimum 1%). Based on 16S rRNA gene sequence analysis, the near phylogenetic neighbours of strain THG-N2.35T were identified as Paracoccus hibisci KACC 18632T (99.0%), Paracoccus tibetensis CGMCC 1.8925T (98.7%), Paracoccus beibuensis CGMCC 1.7295T (98.2%), Paracoccus aestuarii KCTC 22049T (98.2%), Paracoccus rhizosphaerae LMG 26205T (98.1%), Paracoccus zeaxanthinifaciens ATCC 21588T (97.1%), Paracoccus marcusii DSM 11574T (97.0%). Levels of similarity between strain THG-N2.35T and other Paracoccus species were lower than 97.0%. DNA-DNA hybridization values between strain THG-N2.35T and P. hibisci KACC 18632T, P. tibetensis CGMCC 1.8925T, P. beibuensis CGMCC 1.7295T, P. aestuarii KCTC 22049T, P. rhizosphaerae LMG 26205T, P. zeaxanthinifaciens ATCC 21588T, P.marcusii DSM 11574T were 47.5% (42.3%, reciprocal analysis), 36.1% (32.3%), 24.7% (22.1%), 19.2% (16.3%), 11.3% (8.8%), 11.1% (10.8%), 6.1% (5.8%), respectively. The DNA G+C content of strain THG-N2.35T was 62.3mol%. The polar lipids were diphosphatidylglycerol, phosphatidyl-N-methylethanolamine, phosphatidylethanolamine, phosphatidylglycerol and phosphatidylcholine. The quinone was ubiquinone-10 (Q-10). The major fatty acids were C10:0 3OH, C16:0, C18:0 and C18:1 ω7ϲ. On the basis of the phylogenetic analysis, chemotaxonomic data, physiological characteristics and DNA-DNA hybridization data, strain THG-N2.35T represent a novel species of the genus Paracoccus, for which the name Paracoccus pueri sp. nov. is proposed. The type strain is THG-N2.35T (=KACC 18934T=CCTCC AB 2016177T).

Actinotalea solisilvae sp. nov., isolated from forest soil and emended description of the genus Actinotalea.

A Gram-stain-positive, aerobic, non-motile and short-rod-shaped actinobacterium, designated THG-T121T, was isolated from forest soil. Growth occurred at 10-40 °C (optimum 28-30 °C), at pH 6-8 (optimum 7) and at 0-4 % NaCl (optimum 1 %). Based on 16S rRNA gene sequence analysis, the nearest phylogenetic neighbours of strain THG-T121T were identified as Actinotalea ferrariae KCTC 29134T (97.9 %), Actinotalea fermentans KCTC 3251T (97.3 %), Cellulomonas carbonis KCTC 19824T (97.2 %). 16S rRNA gene sequence similarities among strain THG-T121T and other recognized species were lower than 97.0 %. The polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, two phosphatidylinositol mannosides, one unidentified phospholipid, three unidentified glycolipids and one unidentified lipid. The isoprenoid quinone was menaquinone (MK-10(H4)). The major fatty acids were anteiso-C15 : 0, anteiso-C15 : 1 A, C16 : 0, iso-C16 : 0, anteiso-C17 : 0 and iso-C17 : 0. The whole-cell sugars of strain THG-T121T were rhamnose, ribose, mannose and glucose. The peptidoglycan type of strain THG-T121T is A4β, containing l-Orn-D-Ser-L-Asp. The DNA G+C content of strain THG-T121T was 72.4 mol%. DNA-DNA hybridization values between strain THG-T121T and A. ferrariae KCTC 29134T, A. fermentans KCTC 3251T and C. carbonis KCTC 19824T were 30.2 % (27.3 %, reciprocal analysis), 28.4 %, (17.3 %) and 16.9 %, (9.3 %), respectively. On the basis of the phylogenetic analysis, chemotaxonomic data, physiological characteristics and DNA-DNA hybridization data, strain THG-T121T represents a novel species of the genus Actinotalea, for which the name Actinotaleasolisilvae sp. nov. is proposed. The type strain is THG-T121T (=KACC 19191T=CGMCC 4.7389T).

Nonomuraea lycopersici sp. nov., isolated from the root of tomato plants (Solanum lycopersicum L.).

Two novel bacterial strains were isolated from the roots of tomato plants (Solanum lycopersicum L.) from Dengfeng, Henan Province, China. Phylogenetic analysis of 16S rRNA gene sequences showed that the isolates NEAU-DE8(1)T and NEAU-HE1(2) are closely related to one another (99.6% similarity) and are closely relate to Nonomuraea zeae NEAU-ND5T (98.3, 98.3%) and Nonomuraea gerenzanensis DSM 100948T (98.3, 98.0%). Phylogenetic trees constructed based on 16S rRNA gene sequences revealed that both strains grouped with N. zeae NEAU-ND5T within the clade comprising the members of the genus Nonomuraea. Moreover, key chemotaxonomic properties including major menaquinones, fatty acid composition and phospholipid profiles also confirmed the affiliation of the two strains to the genus Nonomuraea. However, DNA-DNA hybridization data, together with physiological and morphological properties, showed that two isolates could be readily distinguished from their close phylogenetic relatives. Thus, on the basis of these phenotypic and genotypic data, strains NEAU-DE8(1)T and NEAU-HE1(2) are concluded to represent a novel species of the genus Nonomuraea, for which the name Nonomuraea lycopersici sp. nov. is proposed. The type strain is NEAU-DE8(1)T (=CGMCC 4.7412T=DSM 104642T).