Paenibacillus thermotolerans sp. nov., isolated from a hot spring in Yunnan Province, south-west China.

Two Gram-positive, rod-shaped, endospore-forming strains, YIM B05601 and YIM B05602T, were isolated from soil sampled at Hamazui hot spring, Tengchong City, Yunnan Province, PR China. Phylogenetic analysis based on 16S rRNA gene sequences suggested that the two strains fell within the genus Paenibacillus, appearing most closely related to Paenibacillus alkalitolerans YIM B00362T (96.9 % sequence similarity). Genome-based phylogenetic analysis confirmed that strains YIM B05601 and YIM B05602T formed a distinct phylogenetic cluster within the genus Paenibacillus. The average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values of strains YIM B05601 and YIM B05602T with the related species P. alkalitolerans YIM B00362T were within the ranges of 74.43-74.57 % and 12.1-18.5 %, respectively, which clearly indicated that strains YIM B05601, YIM B05602T represented a novel species. Strains YIM B05601 and YIM B05602T exhibited 99.6 % 16S rRNA gene sequence similarity. The ANI and dDDH values between the two strains were 99.8 and 100 %, respectively, suggesting that they belong to the same species. Optimum growth for both strains occurred at pH 7.0 and 45 °C. The diagnostic diamino acid in the cell-wall peptidoglycan of strains YIM B05601 and YIM B05602T was meso-diaminopimelic acid. MK-7 was the predominant menaquinone. The polar lipids of strain YIM B05602T were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylmonomethylethanolamine, four unidentified glycolipids, an unidentified polarlipid and phosphatidylinositol mannoside. The major fatty acids of the two stains were iso-C15 : 0 and anteiso-C15 : 0. Based on phylogenomic and phylogenetic analyses coupled with phenotypic and chemotaxonomic characterizations, strains YIM B05601 and YIM B05602T could be classified as a novel species of the genus Paenibacillus, for which the name Paenibacillus thermotolerans sp. nov. is proposed. The type strain is YIM B05602T (=CGMCC 1.60051T=KCTC 43460T=NBRC 115924T).

Parapedobacter tibetensis sp. nov., isolated from shore soil of saline lake in Tibet of China and its genome mining of secondary metabolite.

Five aerobic, Gram-stain-negative, non-motile, non-spore-forming, short rod bacteria strains, designated as C3-1-R+6T, C3-2-M9, B3-2-R-7, B3-2-R-21 and C3-2-M2, were isolated from shore soil of LungmuCo Lake in Tibet of China. The 16S rRNA gene sequence comparisons confirmed their affiliation to the genus Parapedobacter of the family Sphingobacteriaceae, and showed that they were most closely related to Parapedobacter lycopersici KACC 18788T with 94.26 % similarities. The average nucleotide identity (ANI), average amino acid identity (AAI) and digital DNA-DNA hybridization (dDDH) values between them and the validly published Parapedobacter species were all below the thresholds for delineating species, supporting that they were novel species of genus Parapedobacter. The ANI, AAI and dDDH values between strains C3-1-R+6T and Parapedobacter lycopersici KACC 18788T were 72, 75, and 18% respectively. Meanwhile, the ANI/AAI and dDDH values between these five isolates were higher than the threshold values, showing that they belonged to the same species of Parapedobacter. According to genome comparison, the novel isolates have some special biosynthetic gene clusters of secondary metabolites including bacteriton, aryl-polyene, lantipeptide and t1pks, which were absent from their most related phylogenetic neighbours P. lycopersici KACC 18788T and P. pyrenivorans CGMCC 1.12195T. The main polar lipids contained phosphatidylethanolamine, one unidentified phospholipid, one unidentified aminolipid, one unidentified glycolipid and five unidentified lipids. The predominant respiratory quinone was MK-7. The major cellular fatty acids were iso-C15 : 0, summed feature 3 (C16 : 1 ω7c and/or C16 : 1 ω6c) and iso-C17 : 0 3-OH. The genome size of strain C3-1-R+6T was 5 984 948 bp, and its genomic DNA G+C content was 46.21 mol%. To sum up, the five strains were identified as a novel species of the genus Parapedobacter, for which the name Parapedobacter tibetensis sp. nov. was proposed. The type strain was C3-1-R+6T (=CGMCC 1.19194T=KCTC 92150T).

Aquiflexum gelatinilyticum sp. nov., isolated from river water.

Two Gram-stain-negative, strictly aerobic, rod-shaped, non-motile and non-gliding bacteria, designated as XJ19-10T and XJ19-11, were isolated from river water in Xinjiang Uygur Autonomous Region, PR China. Cells of these strains were catalase-, oxidase- and gelatinase-positive and contained carotenoids but no flexirubins. Growth occurred at 10-30 °C, pH 7.0-9.0 and with 0-2.5% (w/v) NaCl. On the basis of the results of 16S rRNA gene sequence and genome analyses, the two isolates represented members of the genus Aquiflexum, and the closest relative was Aquiflexum aquatile Z0201T with 16S rRNA gene sequence pairwise similarities of 97.9-98.1%. Furthermore, the average nucleotide identities and digital DNA-DNA hybridization identities between the two isolates and other relatives were all less than 82.9 and 28.2 %, respectively, all below the species delineation thresholds. The results of pan-genomic analysis indicated that the type strain XJ19-10T shared 2813 core gene clusters with other three type strains of members of the genus Aquiflexum, as well as having 623 strain-specific clusters. The major polar lipids were phosphatidylethanolamine, phosphatidylcholine, an unidentified aminolipid and unidentified lipids. The predominant fatty acids (>10% of the total contents) were iso-C15 : 0, iso-C15 : 1G, iso-C17 : 0 3-OH and summed feature 9, and MK-7 was the respiratory quinone. On the basis of the results of phenotypic, physiological, chemotaxonomic and genotypic characterization, strains XJ19-10T and XJ19-11 are considered to represent a novel species, for which the name Aquiflexum gelatinilyticum sp. nov. is proposed. The type strain is XJ19-10T (=CGMCC 1.19385T =KCTC 92266T).

Devosia ureilytica sp. nov., isolated from Kuche River in China.

Two novel strains, designated XJ19-45T and XJ19-1, were isolated from water of Kuche River in Xinjiang Uygur Autonomous Region, China. Their cells were Gram-stain-negative, aerobic and motile rods. The phylogenetic analyses based on 16S rRNA genes and genomes showed that the two isolates belonged to the genus Devosia and the closest relative was Devosia subaequoris HST3-14T. The 16S rRNA genes sequences pairwise similarities, average nucleotide identities, digital DNA-DNA hybridizations and average amino acid identities between type strain XJ19-45T and other relatives were all less than 98.3, 80.3, 23.6 and 85.7 %, respectively, all below the species delineation thresholds. Pan-genomic analysis indicated that the novel isolate XJ19-45T shared 1594 core gene clusters with the 11 closely related type strains in Devosia, and the number of strain-specific clusters was 390. The major cellular fatty acids (>10 %) of the two isolates were summed feature 8, C18 : 1 ω7c 11-methyl and C16 : 0. Diphosphatidylglycerol, phosphatidylglycerol and glycolipids were the major polar lipids, and Q10 was the detected respiratory quinone. Based on the results of phenotypic, physiological, chemotaxonomic and genotypic characterizations, we propose that the isolates represent a novel species, for which the name Devosia ureilytica sp. nov. is proposed. The type strain is XJ19-45T (=CGMCC 1.19388T=KCTC 92263T).

Propioniciclava soli sp. nov., isolated from forest soil, Yunnan, China, and reclassification of the genus Brevilactibacter into the genus Propioniciclava, and Brevilactibacter sinopodophylli, Brevilactibacter flavus, and Brevilactibacter coleopterorum as Propioniciclava sinopodophylli comb. nov., Propioniciclava flava comb. nov., and Propioniciclava coleopterorum comb. nov., respectively.

A Gram-stain-positive, coccus-shaped, facultatively anaerobic, non-motile bacterial strain, designated YIM S02567T, was isolated from a forest soil sample collected from Gejiu City, Yunnan Province, southwest PR China. Growth was observed at 10-45 °C, at pH 6.0-9.5, in the presence of up to 4.0% (w/v) NaCl on R2A medium. The results of 16S rRNA gene sequence similarity analysis showed that strain YIM S02567T was most closely related to the type strain of Brevilactibacter sinopodophylli (95.4%) and Propioniciclava tarda (94.7%), and phylogenetic analysis based on genome data showed that strain YIM S02567T should be assigned to the genus Propioniciclava. The cell-wall diamino acid was meso-diaminopimelic acid. The major cellular fatty acids were identified as anteiso-C15:0 and C16:0, and the major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, and two unidentified glycolipids. The predominant menaquinone was MK-9(H4). The genomic DNA G + C content was 71.2 mol%. Based on the polyphasic taxonomic evidence, strain YIM S02567T is assigned to a novel member of the genus Propioniciclava, for which the name Propioniciclava soli sp. nov., (type strain YIM S02567T = CCTCC AB 2020128T = CGMCC 1.18504T = KCTC 49478T) is proposed. Furthermore, we propose the reclassification of Brevilactibacter as Propioniciclava gen. nov.

Undibacterium crateris sp. nov., isolated from water of crater lake.

A Gram-stain-negative, aerobic, flagellated, non-spore-forming, rod-shaped bacterium, named B2R-29T, was isolated from water collected from a crater lake on Da Hinggan mountain, PR China. Strain B2R-29T was oxidase- and catalase-positive. On the basis of the results of 16S rRNA gene sequence analyses, strain B2R-29T clearly belonged to the family Oxalobacteraceae, class Betaproteobacteria and showed the highest similarity to Undibacterium oligocarboniphilum EM1T (97.4 %) and to the other species of Undibacterium (less than 96.8 %). In the phylogenetic tree, strain B2R-29T formed a clade with U. oligocarboniphilum EM1T and Undibacterium squillarum CMJ-9T, indicating that is a member of the genus Undibacterium. Digital DNA-DNA hybridization and average nucleotide identity analyses were performed and the values between strain B2R-29T and its closely related Undibacterium species were less than 75.1 % and 16.9 %, respectively. The chemotaxonomic data of B2R-29T were as follows: major uniquinone, Q-8; predominant polar lipids, phosphatidylethanolamine, diphosphatidylglycerol and phosphatidylglycerol; major fatty acids, C16 : 0 and summed feature 3 (C16 : 1 ω7c / C16 : 1 ω6c); predominant polyamines, putrescine, 2-hydroxyputrescine and spermidine. The DNA G+C content was 51.7 mol% from the genomic sequencing data. In accordance with the phenotypic, physiological and chemotaxonomic properties mentioned above, strain B2R-29T represents a novel species of the genus Undibacterium for which the name Undibacterium crateris sp. nov. is proposed. The type strain is B2R-29T (=CGMCC 1.13792T=KCTC 72018T).

Alkalibacillus aidingensis sp. nov., an Bacterium Isolated from Aiding Lake in Xinjiang Province, North-West China.

A bacterial strain, Gram staining negative, aerobic, long rod, motile bacterium with flagellum, designated strain YIM 98829T, was isolated from the Aiding Lake in Xinjiang province, North-West China. The isolate produced oval subterminal endospores in swollen sporangia. The predominant menaquinone was MK-7. The cell wall peptidoglycan contained ornithine, serine, aspartic acid, glutamic acid, and alanine, while diaminopimelic acid could not be detected. The major whole-cell sugars contained xylose, glucose, galactose, and mannose. Diphosphatidylglycerol, phosphatidylglycerol, one unknown phospholipid, and two unidentified aminophospholipids were part of the polar lipid profile. Iso-C15:0 and anteiso-C15:0 were the major fatty acids. The DNA G + C content of the type strain was 38.0 mol%. Phylogenetic analysis indicated that the isolate belongs to the genus Alkalibacillus. However, it differed from its closest relatives, A. haloalkaliphilus DSM 5271T (97.04%), A. filiformis 4AGT (96.99%), and A. silvisoli BM2T (96.95%) in some physiological characteristics. DNA-DNA hybridization result indicated low levels of relatedness between strain YIM 98829T and A. haloalkaliphilus JCM 12303T (16.9%). On the basis of physiological, phenotypic, and chemotaxonomic data, strain YIM 98829T represents a novel species of genus Alkalibacillus, for which the name Alkalibacillus aidingensis sp. nov. is proposed. The type strain is YIM 98829T (= NBRC 114103T = CGMCC 1.17260T = DSM 112470T).

Phenotypic divergence of thermotolerance: Molecular basis and cold adaptive evolution related to intrinsic DNA flexibility of glacier-inhabiting Cryobacterium strains.

The link between guanine-cytosine (GC) content and thermal adaptation is controversial. Here, we compared maximum growth temperature (TMGT ) and genomics of 78 Cryobacterium strains to avoid unreliable conclusions resulting from distantly phylogenetic groups. Phylogenomic analysis revealed this taxon had much higher diversification than we knew. Interestingly, these strains showed thermotolerance divergence with phylogenetic cohesion. A significant difference was found between TMGT ≤ 20°C strains and TMGT > 20°C strains in genomic GC content which mainly caused by variation of GC3. TMGT ≤ 20°C strains tended to use synonymous codons ended with A/U, but TMGT > 20°C strains tended to use G/C. Lower GC content at synonymous sites (≈GC3) of TMGT ≤ 20°C strains could provide lower intrinsic DNA flexibility which strongly associated with optimal molecular dynamics, and then guarantee DNA function at lower growth temperatures. This analysis of codon bias revealed close relationships for thermal adaptation, GC content at synonymous sites (≈GC3), intrinsic DNA flexibility and optimal DNA dynamics. Natural selection was main force driving this codon bias; strains with lower TMGT endured stronger natural selection. Therefore, this study provided molecular basis for bacterial adaptive evolution from moderate temperature to low temperature.

Genomic characterization of closely related species in the Rumoiensis clade infers ecogenomic signatures to non-marine environments.

Members of the family Vibrionaceae are generally found in marine and brackish environments, playing important roles in nutrient cycling. The Rumoiensis clade is an unconventional group in the genus Vibrio, currently comprising six species from different origins including two species isolated from non-marine environments. In this study, we performed comparative genome analysis of all six species in the clade using their complete genome sequences. We found that two non-marine species, Vibrio casei and Vibrio gangliei, lacked the genes responsible for algal polysaccharide degradation, while a number of glycoside hydrolase genes were enriched in these two species. Expansion of insertion sequences was observed in V. casei and Vibrio rumoiensis, which suggests ongoing genomic changes associated with niche adaptations. The genes responsible for the metabolism of glucosylglycerate, a compound known to play a role as compatible solutes under nitrogen limitation, were conserved across the clade. These characteristics, along with genes encoding species-specific functions, may reflect the habit expansion which has led to the current distribution of Rumoiensis clade species. Genome analysis of all species in a single clade give us valuable insights into the genomic background of the Rumoiensis clade species and emphasize the genomic diversity and versatility of Vibrionaceae.

Craterilacuibacter sinensis gen. nov. sp. nov., isolated from a crater lake in China.

Two bacterial strains, designated B2N2-7T and B2N2-12, were isolated from Buteha crater lake in the Greater Khingan Mountain of China. The two strains were Gram-stain-negative, non-spore-forming, motile with a single polar flagellum, short rod-shaped bacteria. They were catalase- and oxidase-positive. Optimal growth occurred at 20-25 ℃, at pH 7.5-8.0 and with 0-1.0 % (w/v) NaCl. Based on phylogenomic analysis, strains B2N2-7T and B2N2-12 were assigned to the family Neisseriaceae, and their 16S rRNA gene sequences showed the highest similarities to that of Aquitalea denitrificans 5YN1-3T (<94.2 %). The predominant cellular fatty acids were C16 : 0 and summed feature 3 (comprising C16 : 1ω7c/C16 : 1 ω6c). The major respiratory quinone was ubiquinone 8 (Q-8). The polar lipids were phosphatidylglycerol (PG), diphosphatidylglycerol (DPG), phosphatidylethanolamine (PE), two unidentified aminophospholipids (APL) and some unidentified lipids (L). The genomic DNA G+C content of strain B2N2-7T was 59.4 mol% according to the genomic sequencing result. Based on the phylogenetic, genotypic and chemotaxonomic analyses, the two strains are proposed to represent a novel species of a new genus in the family Neisseriaceae, named Craterilacuibacter sinensis gen. nov., sp. nov. The type strain of Craterilacuibacter sinensis is B2N2-7T (=CGMCC 1.17189T=KCTC 73735T); B2N2-12 (=CGMCC 1.17190=KCTC 72734) is a second strain of the species.

Cryobacterium ruanii sp. nov. and Cryobacterium breve sp. nov., isolated from glaciers.

Strains Sr36T and TMT4-23T were isolated from No. 1 glacier in Xinjiang Uygur Autonomous Region and Toumingmengke glacier in Gansu Province, PR China, respectively. They were Gram-stain-positive and rod-shaped micro-organisms. The optimum growth temperature of the two strains was 10-14 °C. Phylogenetic analysis showed that the two strains were related to members of the genus Cryobacterium. The average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values between strain Sr36T and its close relatives Cryobacterium luteum Hh15T, Cryobacterium aureum Hh31T, Cryobacterium levicorallinum Hh34T and Cryobacterium flavum Hh8T were 81.16-87.24 and 28.0-32.5 %, respectively. The ANI and dDDH values between strain TMT4-23T and its close relative Cryobacterium psychrotolerans 0549T were 81.16 and 22.3 %. The polar lipids of strain Sr36T were diphosphatidylglycerol, phosphatidylglycerol, one unidentified glycolipid and three unidentified lipids. The polar lipids of strain TMT4-23T were diphosphatidylglycerol, phosphatidylglycerol, one unidentified glycolipid, one unidentified phospholipid and six unidentified lipids. The major fatty acids of strain Sr36T were anteiso-C15 : 0, iso-C16 : 0, anteiso-C17 : 0 and anteiso-C15 : 1. The major fatty acids of strain TMT4-23T were anteiso-C15 : 0, anteiso-C17 : 0, iso-C16 : 0, anteiso-C15 : 1 and iso-C15 : 1. Both strains contained 2,4-diaminobutyric acid and their predominant menaquinone was MK-10. On the basis of the phenotypic, phylogenetic and genotypic data, two novel species Cryobacterium ruanii sp. nov. (type strain = Sr36T=CGMCC 1.9275T=NBRC 113797T) and Cryobacterium breve sp. nov. (type strain =TMT4-23T=CGMCC 1.9556T=NBRC 113800T) are proposed.

Naganishia floricola sp. nov., a novel basidiomycetous yeast species isolated from flowers of Sorbaria sorbifolia.

Two yeast strains representing a novel species in the basidiomycetous yeast genus Naganishia were isolated from flowers of Sorbaria sorbifolia collected in Beijing Olympic Forest Park, PR China. Results of multi-gene phylogenetic analysis indicated that the two strains were closely related to the type strains of Naganishia bhutanensis (CBS 6294T) and Naganishia antarctica (CBS 7687T). However, the new isolates differed from N. bhutanensis CBS 6294T by 1.79 % sequence divergence in the D1/D2 domain (11 nt substitutions and three indels), and 2.42 % (15 nt differences and one indel) to N. antarctica CBS 7687T. In the ITS region, the new isolates showed 1.15 % divergence (7 nt substitutions and one indel) to N. bhutanensis CBS 6294T and 0.92 % divergence (5 nt substitutions and no indels) to N. antarctica CBS 7687T. A phylogenetic analysis employing the sequences of six genes (D1/D2 domain of large subunit rDNA, ITS, small subunit rDNA, two subunits of the RNA polymerase II and elongation factor-1α) indicated that the novel species belonged to the genus Naganishia and formed a well-supported clade with N. bhutanensis, N. antarctica and N. indica. Moreover, the two strains differed from their closest relatives by the ability to grow on distinct carbon and nitrogen sources and ability to grow at 30 °C. On the basis of these findings, we propose a novel species in the genus Naganishia (Filobasidiales), Naganishia floricola sp. nov. (holotype CGMCC 2.5856).

Flavobacterium psychrotolerans sp. nov., a psychrotolerant bacterium isolated from Renlongba glacier on the Tibetan Plateau.

One aerobic, Gram-stain-negative, rod-shaped bacterium, designated strain RB1R5T, was isolated from Renlongba glacier in Tibet Autonomous Region, China. Growth was observed at 4-25 °C and pH 7.0-8.0. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain RB1R5T belongs to the genus Flavobacterium, and its closest relatives are Flavobacterium luteum IMCC26026T and Flavobacterium psychroterrae CCM 8827T with 97.75 and 97.42% gene sequence similarities, respectively. The average nucleotide identity and DNA-DNA hybridization values between strain RB1R5T and its closest relative Flavobacterium luteum IMCC26026T were 78.02 and 21.8 %, respectively. The major cellular fatty acids were anteiso-C15 : 0 and summed feature 3 (comprising C16:1ω7c/C16:1ω6c). The sole menaquinone was MK-6. The major phospholipid was phosphatidylethanolamine. On the basis of phenotypic, phylogenetic and chemotaxonomic characterizations, strain RB1R5T represents a novel species of the genus Flavobacterium, for which the name Flavobacterium psychrotolerans sp. nov. is proposed. The type strain is RB1R5T (=CGMCC 1.23024T=NBRC 113060T).

Flavobacterium restrictum sp. nov., Flavobacterium rhamnosiphilum sp. nov., and Flavobacterium zepuense sp. nov. isolated from glaciers.

Three strains, designated as LB1R34T, LB3P52T and ZT4R6T, were isolated from glaciers located on the Tibetan Plateau, PR China. The strains were Gram-stain-negative, aerobic, rod-shaped, non-motile and yellow. Phylogenetic analysis based on the 16S rRNA gene and genomic sequences indicated that they were related to the members of the genus Flavobacterium. The 16S rRNA gene sequences similarities between the three strains were 92.31-96.93 %. The average nucleotide identity values and digital DNA-DNA hybridization values between these three strains and their closest relatives were 76.80-91.33 % and 21.3-44.2 %, respectively. LB1R34T, LB3P52T and ZT4R6T contained MK-6 as the major menaquinone, summed feature 3 (comprising C16 : 1ω7c and/or C16 : 1ω6c) and iso-C15 : 0 as the major fatty acids. Phosphatidylethanolamine was present in their polar lipids profiles. On the basis of the phenotypic characteristics, he results of phylogenetic analysis and genotypic data, three novel species, Flavobacterium restrictum sp. nov. (type strain=LB1 R34T=CGMCC 1.11493T=NBRC 113650T), Flavobacterium rhamnosiphilum sp. nov. (type strain=LB3 P52T=CGMCC 1.11446T=NBRC 113776T) and Flavobacterium zepuense sp. nov. (type strain=ZT4 R6T=CGMCC 1.11919T=NBRC 113653T) are proposed.

Polymorphobacter arshaanensis sp. nov., containing the photosynthetic gene pufML, isolated from a volcanic lake.

An aerobic, Gram-stain-negative, yellowish, rod-shaped bacterium, designated DJ1R-1T, was isolated from water sample from a volcanic lake, located on Da Hinggan Ling Mountain, PR China. Growth of DJ1R-1T optimally occurred at pH 7.0, at 22-25 °C and with 0-0.5 % (w/v) NaCl concentration. Phylogenetic analysis of 16S rRNA gene sequences indicated that DJ1R-1T was clustered into the genus Polymorphobacter, and showed 96.5 %, 95.9 % and 95.6 % similarities to Polymorphobacter fuscus D40PT, Polymorphobacter multimanifer 262-7T and Polymorphobacter glacialis B555-2T, respectively. The predominant polar lipids were phosphatidylethanolamine, phosphatidylglycerol, one unidentified aminophospholipid, three unidentified aminolipids and one unidentified phospholipid. The major fatty acids were summed feature 8 (C18 : 1ω7c / C18 : 1ω6c, 40.0 %), summed feature 3 (C16 : 1ω7c / C16 : 1ω6c, 25.6 %) and C16 : 0 (13.7 %). The respiratory quinone was ubiquinone-10. The DNA G+C content was 65.0 % according to the genomic sequencing results. On the basis of the results of the phylogenetic analysis, physiological and biochemical properties comparisons, DJ1R-1T was proposed to represent a novel species of the genus Polymorphobacter, with the name Polymorphobacter arshaanensis. The type strain is DJ1R-1T (=CGMCC 1.13788T=KCTC 72014T).

Pseudomonas Saliphila sp. nov., a Bacterium Isolated from Oil-Well Production Water in Qinghai Oilfield of China.

Strain 16W4-4-3 T was isolated from the oil-well production water in Qinghai Oilfield, China. Cells were Gram-stain-negative, rod-shaped, catalase- and oxidase-positive, facultatively anaerobic and motile by single polar flagellum. The 16S rRNA gene sequences of strain 16W4-4-3 T showed the highest similarities with Pseudomonas profundi M5T (98.8%), P. pelagia CL-AP6T (98.0%), P. salina XCD-X85T (97.7%), and P. sabulinigri J64T (97.5%). The phylogenetic trees based on multilocus sequence analyses with concatenating 16S rRNA, gyrB, rpoD and rpoB genes suggested that this strain should be affiliated to the genus Pseudomonas but remotely related from other species. In addition, whole genome analyses revealed that the digital DNA-DNA hybridization values and average nucleotide identities of strain 16W4-4-3 T against its close relatives were all below 28.8% and 86.5%, respectively. Furthermore, the isolate had totally different whole cell protein profile as compared to those of other species. Major fatty acids were summed feature 8 (C18:1ω7c and/or C18:1ω6c), C16:0, summed feature 3 (C16:1ω7c and/or C16:1ω6c) and C17:0cyclo. Major isoprenoid quinone was ubiquinone (Q-9), and major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, and phosphatidylethanolamine. The DNA G + C content was 58.5 mol%. Therefore, phenotypic, phylogenetic, genomic, chemotaxonomic, and proteomic traits showed that the isolate represented a novel species of the genus Pseudomonas, the name Pseudomonas saliphila sp. nov. is proposed. Type strain is 16W4-4-3 T (= CGMCC 1.13350 T = KCTC 72619 T).

Stenotrophobium rhamnosiphilum gen. nov., sp. nov., isolated from a glacier, proposal of Steroidobacteraceae fam. nov. in Nevskiales and emended description of the family Nevskiaceae.

A Gram-stain-negative strain, designated GT1R17T, was isolated from an ervoconite sample collected from Gawalong glacier in the Tibet Autonomous Region, PR China. Strain GT1R17T was catalase- and oxidase-positive, and grew optimally at 20-25°C and pH 7.0. The highest level of 16S rRNA gene sequence similarities were found to members of the genera Nevskia(92.27-93.15 %) and Hydrocarboniphaga(91.92-92.96 %). Phylogenetic analyses based on 16S rRNA gene sequences and genomic data revealed that the strain GT1R17T belonged to the family Nevskiaceae, but could not be assigned to any known genera. The genomic DNA G+C content was 54.4 mol%. The major fatty acids were summed feature 3 (C16 : 1ω6c and/or C16 : 1ω7c), summed feature 8 (C18 : 1 ω7c and/or C18 : 1ω6c), C16 : 0, iso-C16 : 0 and summed feature 2 (C14 : 0 3OH and/or iso-C16 : 1 I). The polar lipids were phosphatidylglycerol, phosphatidylethanolamine and one unidentified lipid. The ubiquinone was Q-8. On the basis of the phenotypic, chemotaxonomic, genotypic and phylogenetic data, a novel species of a new genus,Stenotrophobiumrhamnosiphilum gen. nov., sp. nov. within the family Nevskiaceae, is proposed, with GT1R17T (=CGMCC 1.16137T=NBRC 113346T) as the type strain. In addition, phylogenetic analyses revealed that Steroidobacter and Povalibacter formed an independent clade in the order Nevskiales and were away from the families Nevskiaceae, Algiphilaceae and Salinisphaeraceae. Therefore, we propose to remove Steroidobacter and Povalibacter from the family Nevskiaceae and propose a new family Steroidobacteraceae in the order Nevskiales.

Flavobacterium ranwuense sp. nov., isolated from glacier.

A Gram-stain-negative, aerobic, rod-shaped, devoid of flagella, orange-yellow-coloured bacterium (strain LB2P22T) was isolated from an ice sample collected from the Laigu glacier on the Tibetan Plateau, PR China. The results of 16S rRNA gene sequence analysis indicated that strain LB2P22T belongs to the genus Flavobacterium with highest similarity to Flavobacterium laiguense LB2P30T (98.14 %), Flavobacterium glaciei CGMCC 1.5380T (98.02 %), Flavobacterium psychroterrae CCM 8827T (97.94 %) and Flavobacterium granuli CGMCC 1.10125T (97.91 %). Strain LB2P22T had 77.99-84.99 % average nucleotide identity and 22.3-29.3 % digital DNA-DNA hybridization values with its closest relatives, indicating that it represents a novel species of the genus Flavobacterium. Strain LB2P22T grew at 0-25 °C, pH 7.0-8.0 and up to 1.0 % (w/v) NaCl. The main cellular fatty acids were iso-C15 : 0, summed feature 3 (comprising C16 : 1ω7c/C16 : 1ω6c) and anteiso-C15 : 0. The genomic DNA G+C content is 34.3 mol%. The predominant menaquinone was MK-6 and the polar lipids were phosphatidylethanolamine, one unidentified aminolipid and four unidentified lipids. Based on these data, a novel species, Flavobacteriumranwuense sp. nov., is proposed, with LB2P22T (=CGMCC 1.11361T=NBRC 113777T) as the type strain.

Cryobacterium melibiosiphilum sp. nov., a psychrophilic bacterium isolated from glacier ice.

A psychrophilic, Gram-stain-positive, rod-shaped bacterium, designated Hh39T, was isolated from Xinjiang No. 1 glacier in PR China. Strain Hh39T was catalase-positive, oxidase-negative and could grow at 0-18 °C, pH 6.0-11.0 and in the presence of 0-2.5 % (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain Hh39T belonged to the genus Cryobacterium. The highest level of 16S rRNA gene sequence similarities were found to the type strains of Cryobacterium levicorallinum (99.01 %), Cryobacterium luteum (98.90 %), Cryobacterium aureum (98.90 %) and Cryobacterium roopkundense (98.75 %). However, the low average nucleotide identity (80.65-81.89 %) and digital DNA-DNA hybridization values (22.1-23.8 %) between strain Hh39T and its four closest relatives indicated that it represents a novel species of the genus Cryobacterium. The predominant fatty acids were anteiso-C15:0, anteiso-C15:1, iso-C16:0 and anteiso-C17:0. The major menaquinone was MK-10. The polar lipids were diphosphatidylglycerol, phosphatidylglycerol, one unidentified lipid and one unidentified glycolipid. On the basis of results of phenotypic, genotypic and phylogenetic analyses, a novel species, Cryobacterium melibiosiphilum sp. nov., is proposed, with Hh39T (=NBRC 107884T=CGMCC 1.11212T) as the type strain.

Flavobacterium laiguense sp. nov., a psychrophilic bacterium isolated from Laigu glacier on the Tibetan Plateau.

One bacterial strain, denoted as LB2P30T, was isolated from Laigu glacier located on the Tibetan Plateau, PR China. Strain LB2P30T was an aerobic, non-motile, rod-shaped, yellow-pigmented and Gram-stain-negative bacterium. The temperature range for growth was 4-20 °C (optimum, 14 °C). The phylogenetic analysis based on partial 16S rRNA gene sequences showed that strain LB2P30T belonged to the genus Flavobacterium and was most similar to Flavobacterium fluvii (98.12 %) and Flavobacterium limicola (97.91 %). The average nucleotide identity values between strain LB2P30T and its closest relatives, F. fluvii DSM 19978T and F. limicola DSM 15094T, were 76.73 % and 77.40 %, respectively. Cells of strain LB2P30T contained summed feature 3 (comprising C16:1ω7c and/or C16:1ω6c) and iso-C15 : 0 as the predominant fatty acids and menaquinone-6 as the sole menaquinone. The genomic DNA G+C content was 34.74 mol%. Based on the phenotypic, chemotaxonomic, genotypic and phylogenetic characteristics, we propose strain LB2P30T as a novel species of the genus Flavobacterium with the nomenclature of Flavobacteriumlaiguense sp. nov. The type strain is LB2P30T (=CGMCC 1.11271T=NBRC 113059T).