- Volume 72, Issue 5, 2022

A Gram-stain-negative bacterial strain, designated as E165T, was isolated from a tidal flat sediment of the East China Sea. Strain E165T grew optimally at pH 6, at 32 °C and with 1–2 % (w/v) NaCl. The 16S rRNA gene sequence similarity results revealed that strain E165T was most closely related to Marinomonas rhizomae IVIA-Po-145T, Marinomonas polaris CK13T, Marinomonas foliarum IVIA-Po-155T, Marinomonas hwangdonensis HDW-15T, Marinomonas pontica 46-16T, Marinomonas mangrovi B20-1T and Marinomonas shanghaiensis DSL-35T with values of 97.0–98.5 %. The digital DNA–DNA hybridization and average nucleotide identity values between strain E165T and the reference strains were 21.9–34.3 % and 77.6–87.3 %, respectively. The DNA G+C content of the isolate was 42.9 mol%. Strain E165T contained Q-8 as the sole ubiquinone and C16 : 0, summed feature 8 (C18 : 1  ω7c and/or C18 : 1  ω6c) and summed feature 3 (C16 : 1  ω7c and/or C16 : 1  ω6c) as the major fatty acids. The major polar lipids of strain E165T were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, aminolipid and aminophospholipid. On the basis of phenotypic characteristics, phylogenetic analysis and DNA–DNA relatedness, a novel species, Marinomonas lutimaris sp. nov., is proposed with E165T (=MCCC 1K06241T=KCTC 82809T) as the type strain.

Strain GY_HT was isolated from an artificial wetland in Okcheon, Chungcheongbuk-do Province, Republic of Korea. Strain GY_HT was closely related to Pseudolabrys taiwanensis CC-BB4T based on 16S rRNA gene sequences (94.7 % similarity) and clustered within the family Nitrobacteraceae . Cells of the isolate were Gram-stain-negative, catalase-negative and oxidase-positive, and colonies were white or pale transparent. A flagellum was observed, and the isolate could respire both aerobically and anaerobically. Growth of GY_ HT was observed in the following conditions: 10–45 °C, pH 5–11 and 0–4 % NaCl. The optimal conditions for growth were 25 °C, pH 6.5–7.5 and 0.5–1.5 % NaCl. The major fatty acids were C19 : 0 cyclo ω8c (35.8 %) and summed feature 8 (C18 : 1  ω7c/C18 : 1  ω6c; 27.4 %). The major quinone was found to be ubiquinone-10. Diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol and phosphatidylcholine were the major polar lipids. The G+C content of the genome of GY_HT was 63.3 mol%. Based on its phylogenomic, physiological and biochemical attributes, strain GY_HT represents a novel species of a novel genus of the family Nitrobacteraceae . We propose the name as Undibacter mobilis gen. nov., sp. nov. The type strain is GY_HT (=KCTC 62792T=JCM 32856T).

A Gram-stain-negative, rod-shaped bacterial strain designated as 20VBR1T was isolated from a valley glacier (Vestrebroggerbreen) snout ice sample from Ny-Ålesund, Svalbard, Arctic. The colonies were smooth, circular and light creamish on half-strength R2A agar and grew at 10–35 °C (optimum, 20 °C), at pH 6.5–8.0 (optimum, 7.0) and with 0–2.5 % (w/v) NaCl (optimum, 0.5 %). 16S rRNA gene sequence analysis revealed that strain 20VBR1T belonged to the genus Phenylobacterium and was most closely affiliated to Phenylobacterium aquaticum W2-3-4T (97.65 % similarity), Phenylobacterium haematophilum LMG 11050T (97.57 %) and Phenylobacterium koreense Slu-01T (96.91 %). 20VBR1T has a genome size of 4.24 Mb, comprising 4185 predicted genes with a DNA G+C content of 67.86 mol%. DNA–DNA hybridization experiments indicated that the DNA–DNA relatedness between strain 20VBR1T and P. aquaticum KACC 18306T was 41.95±4.36 %, well below the threshold (<70 %) to delineate bacterial species. Genome relatedness indexes revealed that the average nucleotide identity and digital DNA–DNA hybridization values between 20VBR1T and its closest phylogenomic relative, P. aquaticum KACC 18306T, were 78.97 and 22.10 %, respectively. The predominant isoprenoid quinone was ubiquinone (Q-10) and the major polar lipids were phosphatidylglycerol, one unknown phospholipid, one unknown glycolipid and four unidentified polar lipids. The major fatty acids (>10 %) of strain 20VBR1T were summed feature 8 (comprising C18 : 1 ω7c and/or C18 : 1 ω6c), summed feature 3 (comprising C16 : 1 ω6c and/or C16 : 1 ω7c) and C16 : 0. Based on the physiological, biochemical, chemotaxonomic, phylogenetic and phylogenomic analyses, isolate 20VBR1T is considered to represent a novel species of the genus Phenylobacterium , for which the name Phenylobacterium glaciei sp. nov. is proposed. The type strain is 20VBR1T (=JCM 33227 T=DSM 111428 T=MCC 4220 T).

A Gram-stain-negative, facultatively anaerobic, oxidase-negative and catalase-positive predatory bacillus, designated strain V1718T, was isolated from Xiaoshi Island, PR China. Strain V1718T was found to be closely related to Lujinxingia sediminis SEH01T, with 89.8 % similarity in the 16S rRNA gene sequence, followed by Bradymonas sediminis FA350T with a similarity of 88.4 %. Strain V1718T had the ability to prey on other bacteria, and selective predation on members of Algoriphagus , Nocardioides and Bacillus occurred with the strain. Growth was observed within the range of 20–45 °C (optimal at 37 °C), pH 6.5–9.0 (optimal at pH 8.0) and 1–10 % NaCl (optimal at 3–4 %, w/v). The predominant cellular fatty acids in strain V1718T were iso-C15 : 0 (53.0 %) and C16 : 0 (19.1 %). The major polar lipids present in the strain were phosphatidylglycerol and phosphatidylethanolamine, and the respiratory quinone was menaquinone MK-7. The complete genome sequence of strain V1718T was 5 847 748 bp with a G+C content of 55.2 mol%. The topology of the phylogenomic tree indicated that strain V1718T forms a separate branch in the same clade with the genus Lujinxingia and the family Bradymonadaceae . The average nucleotide identity and average amino acid identity values were 66.4 and 48.6 %, respectively, with Bradymonas sediminis FA350T (type species of Bradymonas ) and 66.8 % and 48.9 % with Lujinxingia litoralis B210T (type species of Lujinxingia ). The genes related to biosynthesis pathways of several important chemical compounds could not be found in the genome of strain V1718T, which was predicted to be the intrinsic reason for predation in this group. The physiological, biochemical and phylogenetic properties of strain V1718T suggest that it belongs to a novel family distinct from other culturable bradymonabacteria. The name Microvenator marinus gen. nov., sp. nov. is proposed, with strain V1718T (=KCTC 72082T=MCCC 1H00380T) as type strain; the name Microvenatoraceae fam. nov. is also proposed. Meanwhile, the genus Lujinxingia can also be taxonomic classified as Lujinxingiaceae fam. nov. Thus, two novel families and a novel genus of the order Bradymonadales are proposed in this paper.

Seven bacterial strains isolated from bovine endocarditis in six animals from different geographic regions were investigated in a polyphasic taxonomic approach. Phylogenetic analysis based on 16S rRNA gene sequences placed all seven isolates on a distinct, monophyletic cluster in the family Neisseriaceae with closest similarity to type strains of Alysiella filiformis (97.06 %) and Kingella kingae (96.34 %). Whole genome sequence analysis of isolates confirmed their species status, with an average nucleotide identity >96 % between isolates and <80 % to other type species of genera of Neisseriaceae while digital DNA–DNA hybridization values were >80 % and<18 %, respectively. The DNA G+C content was 42.5–43.0 mol%. Whole genome sequence based phylogeny showed the isolates being monophyletic and separated from established genera, thereby forming a new genus within the family Neisseriaceae . Similarly, analysis of MALDI-TOF MS reference spectra clustered the isolates close together and clearly separated from other genera, making this the method of choice for identification. Biochemical markers based on classical as well as commercial identification schemes allowed separation from closely related Neisseriaceae genera, even though the new taxon is biochemically not very active. Major fatty acids are C12 : 0, C14 : 0 and C16 : 0. The major quinone is ubiquinone Q-8. In the polar lipid profile, diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and phospholipid were predominant. We propose the novel genus Wielerella with the type species Wielerella bovis gen. nov., sp. nov. The type strain is CCUG 44465T (=DSM 113289T=JF 2483T) isolated post mortem from a cow with endocarditis in Switzerland.

Two strains designated as c1T and c7T, were isolated from the landfill leachate of a domestic waste treatment plant in Huizhou City, Guangdong Province, PR China. The cells of both strains were aerobic, rod-shaped, non-motile and formed yellow colonies on Reasoner’s 2A agar plates. Strain c1T grew at 10–42 °C (optimum, 30 °C), pH 4.5–10.5 (optimum, pH 7.0) and 0–2.0 % (w/v) NaCl (optimum, 0–0.5 %). Strain c7T grew at 10–42 °C (optimum, 30 °C), pH 4.5–10.5 (optimum, pH 6.0) and 0–2.0 % (w/v) NaCl (optimum, 0–0.5 %). Phylogenetic analyses revealed that strains c1T and c7T belong to the genus Novosphingobium . The 16S rRNA gene sequence similarities of strains c1T and c7T to the type strains of Novosphingobium species were 94.5–98.2 % and 94.3–99.1 %, respectively. The calculated pairwise average nucleotide identity values among strains c1T, c7T and the reference strains were in the range of 75.2–85.9 % and the calculated pairwise average amino acid identity values among strains c1T, c7T and reference strains were in the range of 72.0–88.3 %. Their major respiratory quinone was Q-10, and the major cellular fatty acids were C18 : 1  ω7c, C18 : 0, C16 : 1  ω7c, C16 : 0 and C14 : 0 2OH. The major polar lipids of strains c1T and c7T were phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, sphingoglycolipid, unidentified lipids and unidentified phospholipid. Based on phenotypic, chemotaxonomic, phylogenetic and genomic results from this study, strains c1T and c7T should represent two independent novel species of Novosphingobium , for which the names Novosphingobium percolationis sp. nov. (type strain c1T=GDMCC 1.2555T=KCTC 82826T) and Novosphingobium huizhouense sp. nov. (type strain c7T=GDMCC 1.2556T=KCTC 82827T) are proposed. The gene function annotation results of strains c1T and c7T suggest that they could play an important role in the degradation of organic pollutants.

A novel rod-shaped, Gram-stain-negative, aerobic bacterial strain, designated Cra20T, was isolated from the root surface of Leontopodium leontopodioides collected in the Tianshan Mountains, Xinjiang, PR China. Phylogenetic analysis based on 16S rRNA gene sequences, indicated that strain Cra20T was affiliated with the genus Sphingomonas , and was most closely related to Sphingomonas gei ZFGT-11T (99.0 %), Sphingomonas naasensis KIS18-15T (97.8%) and Sphingomonas kyeonggiensis THG-DT81T (97.2 %). The average nucleotide identity values between strain Cra20T, S. gei ZFGT-11T, S. naasensis KIS18-15T and S. kyeonggiensis THG-DT81T were 86.2, 84.2 and 78.2 %, respectively. The genomic DNA G+C content of strain Cra20T was 65.6 mol% (whole genome sequence), and Q-10 was the predominant ubiquinone. The major cellular fatty acids of strain Cra20T were summed feature 8 (comprising C18 : 1  ω6c and/or C18 : 1  ω7c, 67.3 %) and C14 : 0 2-OH (6.4 %). On the basis of genotypic, phenotypic and biochemical data, strain Cra20T is considered to represent a novel species of the genus Sphingomonas , for which the name Sphingomonas psychrotolerans sp. nov. is proposed. The type strain is Cra20T (=CGMCC 1.15510T=NBRC 112697T).

A Gram-stain-negative, aerobic, motile, rod-shaped novel bacterial strain, designated as MA21411-1T, was isolated from the Korean coast. The colonies were white-yellow-coloured, smooth, convex and entire, spherical and 1.0–1.8 mm in diameter. Phylogenetic analysis based on the 16S rRNA gene sequence showed that strain MA21411-1T is closely related to species of the genus Pseudophaeobacter . The 16S rRNA gene sequence similarities between strain MA21411-1T and Pseudophaeobacter arcticus DSM 23566T, Phaeobacter porticola DSM 103148T and Pseudophaeobacter leonis DSM 25627T were 98.31, 97.80 and 97.28 %, respectively. Strain MA21411-1T has a draft genome size of 4 294 042 bp, annotated with 4125 protein-coding genes, and 53 tRNA, three rRNA and one tmRNA genes. The genomic DNA G+C content was 59.2 mol%. Comparative genome analysis revealed that the average nucleotide identity, digital DNA–DNA hybridization and average amino acid identity values among strain MA21411-1T and other related species were below the cut-off levels of 95, 70 and 95.5 %, respectively. The growth temperature range for growth was 15–28 °C (optimum, 25 °C), pH range was 6.0–9.0 (optimum, pH 6.0), and salt tolerance range was 0.5–4 % (optimum 0.5 %). Ubiquinone-10 was the sole quinone present in MA21411-1T and all three closely related strains. The major cellular fatty acid (>10 %) of the strain was summed feature 8 (C18 : 1  ω7c and/or C18 : 1  ω6c). The polar lipid profile contained phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine and four unidentified polar lipids. Based on the phylogenetic tree, as well as phenotypic, chemotaxonomic and genomic features, strain MA21411-1T represents a novel species of the genus Pseudophaeobacter , for which the name Pseudophaeobacter flagellatus sp. nov. is proposed. The type strain is MA21411-1T (=KCTC 92095T=GDMCC 1.2988T).

A Gram-stain-negative, non-spore-forming, motile, aerobic bacterium (strain C21T) was isolated from coral and identified using polyphasic identification approach. Global alignment of 16S rRNA gene sequences indicated that strain C21T shares 95.7 % sequence identity to its closest neighbour, Marinibactrum halimedae NBRC 110095T, followed by other type strains with identities of lower than 95 %. The average nucleotide identity and average amino acid identity values between strain C21T and M. halimedae NBRC 110095T were 69.6 and 64.8 %, respectively, indicating that strain C21T may represent a new species in a new genus. Phylogenetic analysis based on 16S rRNA gene and phylogenomic results indicated that strain C21T forms a distinct branch in the family Cellvibrionaceae . Cellular fatty acids and polar lipids could also readily distinguish strain C21T from closely related type strains. Therefore, strain C21T is suggested to represent a new species in a new genus, for which the name Sessilibacter corallicola gen. nov., sp. nov. is proposed. The type strain is C21T (=MCCC 1K03260T=KCTC 62317T).

Two strains, TMPB967T and TTPB476, were isolated from two different locations in the Mariana Trench. Cells of strains TMPB967T and TTPB476 were Gram-negative, curved rod-shaped (0.35–0.6 µm×2–4 µm) with flagella. Both strains were catalase- and oxidase-positive. Strains TMPB967T and TTPB476 could grow at 4–37 °C (optimum, 37 °C), at pH 6–9 (optimum, pH 6–7) and in the presence of 0–8 % (w/v) NaCl (optimum, 5 %). Both strains could grow with tetradecane or hexadecane as the sole carbon source. The predominant isoprenoid quinone was ubiquinone 9. The major cellular fatty acids of strains TMPB967T and TTPB476 were C18 : 1  ω9c, C16 : 0 and summed feature 3 (C16 : 1  ω7c or ω6c). The polar lipid profile included phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and an unknown aminolipid. The DNA G+C contents of strains TMPB967T and TTPB476 were 53.1 and 53.0 mol%, respectively. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the most closely related validly published species were Thalassolituus marinus IMCC1826T (97.1 % similarity) and Thalassolituus oleivorans MIL-1T (95.9 % similarity). Digital DNA–DNA hybridization results of strain TMPB967T with TTPB476, T. marinus IMCC1826T and T. oleivorans MIL-1T were 99.9, 20.9 and 20.2 %, respectively. Average nucleotide identity results of strain TMPB967T with TTPB476, T. marinus IMCC1826T and T. oleivorans MIL-1T were 100, 75.8 and 72.0 %, respectively. On the basis of the phenotypic, chemotaxonomic and molecular features, strains TMPB967T and TTPB476 belong to a novel species within the genus Thalassolituus , for which the name Thalassolituus alkanivorans sp. nov. is proposed. The type strain is TMPB967T (=KCTC 82621T=MCCC 1K05476T).

Two strains representing a novel yeast species were isolated from plant leaves collected in the Baotianman Nature Reserve in Henan Province, central China. Phylogenetic analysis based on the concatenated sequences of the internal transcribed spacer (ITS) region (ITS1-5.8S–ITS2) and the D1/D2 domain of the large subunit rRNA gene revealed that the novel species belonged to the genus Hyphopichia, although the formation of ascospores was not observed. The novel species was related most closely to Hyphopichia paragotoi CBS 13913T but they differed by 0.9 % sequence divergence (five substitutions) in the D1/D2 domain and by 3.7 % sequence divergence (seven substitutions and eight gaps) in the ITS region. Furthermore, the novel species can also be differentiated from the closely related species in some biochemical and physiological characteristics. The species name of Hyphopichia xiaguanensis f.a., sp. nov. (Holotype CBS 16668, Mycobank MB 842425) is proposed to accommodate strains NYNU 20899T and NYNU 20914.

The taxonomic assignment of Brevibacterium frigoritolerans together with the in-house environmental isolate EB93 was reassessed in this study using phylogenetic and phylogenomic approaches, and the detection of multiple molecular synapomorphies. Results from the reconstructed phylogenetic trees based on the 16S rRNA gene sequences, the concatenated protein sequences of GyrA-GyrB-RpoB-RpoC, and the whole-genome sequences revealed the consistent exclusion of B. frigoritolerans and the environmental isolate EB93 from the cluster formed by the type strains of the genus Brevibacterium . In addition, B. frigoritolerans and the environmental isolate EB93 were both observed to form a clade together with the type strains of the genus Peribacillus . The results from the analysis of the digital DNA–DNA hybridization, average nucleotide identity, average amino acid identity and the difference in the G+C content also corroborated with the phylogenetic inference, and that B. frigoritolerans and the environmental isolate EB93 were of the same species. Furthermore, the presence of the molecular synapomorphies in the protein sequences noted in the description of the genus Peribacillus were also observed in B. frigoritolerans , further strengthening its taxonomic affiliation in the genus. Based on the evidence from the multiple lines of analyses, we propose the reclassification of Brevibacterium frigoritolerans as a member of the genus Peribacillus and assume the name Peribacillus frigoritolerans comb. nov. (type strain DSM 8801 T=ATCC 25097T=CCUG 43489T=CIP 67.20T=JCM 11681T).

The species Blastobotrys navarrensis Sesma and Ramirez was delineated based on the description of the single strain CBS 139.77T. Based on its phenotypic similarities to Blastobotrys proliferans, B. navarrensis CBS 139.77T was later considered a synonym of B. proliferans. In the present study, we isolated the yeast strain IST 508 (=PYCC 8784=CBS 16671) from the soil surrounding an olive tree in Ferreira do Alentejo, Portugal. The phylogenetic analysis of D1/D2 domain and ITS sequences from strain IST 508 indicates that is closely related to B. navarrensis and B. proliferans. Although strain IST 508 differs from B. navarrensis CBS 139.77T by 14 substitutions and 20 indels (6.6 % divergence) in the ITS sequence, no divergence was detected at the level of D1/D2 domain, mitochondrial small subunit rDNA, and cytochrome oxidase II sequences. On the other hand, strains IST 508 and CBS 139.77 differ from B. proliferans NRRL Y-17577T by eight substitutions (1.4 % divergence) in the D1/D2 domain sequence, by 16 substitutions (2.7 % divergence) in the cytochrome oxidase II sequence, and by 16 substitutions (3.7 % divergence) in the mitochondrial small subunit rDNA sequence. Due to the high number of variable phenotypic tests in B. proliferans and B. navarrensis, strains from the two species are difficult to distinguish. Contrasting with what is described for other Blastobotrys species, no differences were detected at the level of micromorphology between the two species. Nevertheless, based on the molecular differences between the two strains, CBS 139.77 and IST 508, and B. proliferans NRRL Y-17577T and their phylogenetic analysis, strains CBS 139.77 and IST 508 are from B. navarrensis and this species should be considered as an independent species and not a later synonym of B. proliferans. We propose an emended description of B. navarrensis.