Pairwise 16S rRNA Gene Sequence Research Articles

Psychromicrobium Xiongbiense sp. nov., an Actinobacterium Isolated from Forest Soil.

A novel Gram-stain-positive, oval-shaped, and non-flagellated bacterial strain YIM S02556T was isolated from forest soil in Xiongbi Town, Shizong County, Qujing City, Yunnan Province, southwestern China. The strain exhibited high pairwise 16S rRNA gene sequence similarity with Psychromicrobium lacuslunae (97.3%) and Psychromicrobium silvestre (96.3%). Strain YIM S02556T exhibited an average nucleotide identity (ANI) of 72.5% with P. lacuslunae IHBB 11,108T and 72.8% ANI with P. silvestre AK 20-18T. The digital DNA-DNA hybridization (dDDH) value between strain YIM S02556T and P. lacuslunae IHBB 11,108T was 20.2%, while with P. silvestre AK 20-18T, the dDDH value was 20.8%. Strain YIM S02556T exhibited optimal growth at 28°C, pH 7.0, without NaCl. Growth occurred within 10-37 ℃, pH 5.0-8.0, and in the presence of up to 5% w/v NaCl concentration. The genome size was 3.1 Mbp with 64.2% G + C content. The predominant menaquinone was MK-8(H4). The major cellular fatty acid was anteiso-C15:0. Based on the polyphasic analysis, strain YIM S02556T (= KCTC 49,805T = CCTCC AB2020166T) represents a novel Psychromicrobium species in which the name Psychromicrobium xiongbiense sp.nov. was proposed.

Microbulbifer bruguierae sp. nov., isolated from sediment of mangrove plant Bruguiera sexangula, and comparative genomic analyses of the genus Microbulbifer.

A Gram-negative, non-motile, strictly aerobic, rod-shaped bacterium, designated as H12T, was isolated from the sediments of mangrove plant Bruguiera sexangula taken from Dapeng district, Shenzhen, PR China. The pairwise 16S rRNA gene sequence analysis showed that strain H12T shared high identity levels with species of the genus Microbulbifer, with the highest similarity level of 98.5 % to M. pacificus SPO729T, followed by 98.1 % to M. donghaiensis CN85T. Phylogenetic analysis using core-genome sequences showed that strain H12T formed a cluster with type species of M. pacificus SPO729T and M. harenosus HB161719T. The complete genome of strain H12T was 4 481 396 bp in size and its DNA G+C content was 56.7 mol%. The average nucleotide identity and digital DNA-DNA hybridization values among strain H12T and type species of genus Microbulbifer were below the cut-off levels of 95-96 and 70 %, respectively. The predominant cellular fatty acids of strain H12T were iso-C15 : 0 (22.5 %) and C18 : 1 ω7c (13.9 %). Ubiquinone-8 was detected as the major respiratory quinone. The polar lipids of strain H12T comprised one phosphatidylglycerol, one phosphatidylethanolamine, one unidentified aminoglycophospholipid, one unidentified glycophospholipid, three unidentified glycolipids, two unidentified aminolipids, and one unidentified lipid. Based on polyphasic evidence, strain H12T represents a novel species of the genus Microbulbifer, for which the name Microbulbifer bruguierae sp. nov. is proposed. The type strain is H12T (=KCTC 92859T=MCCC 1K08451T). Comparative genomic analyses of strain H12T with strains of the genus Microbulbifer reveal its potential in degradation of pectin.

Genomic insight into Myroides oncorhynchi sp. nov., a new member of the Myroides genus, isolated from the internal organ of rainbow trout (Oncorhynchus mykiss).

The strain M-43T was isolated from the Oncorhynchus mykiss from a fish farm in Mugla, Turkey. Pairwise 16S rRNA gene sequence analysis was used to identify strain M-43T. The strain was a member of the genus Myroides sharing the highest 16S rRNA gene sequence identity levels of 98.7%, 98.3%, and 98.3% with the type strains of M. profundi D25T, M. odoratimimus subsp. odoratimimus CCUG 39352T and M. odoratimimus subsp. xuanwuensis DSM27251T, respectively. A polyphasic taxonomic approach including whole genome-based analyses was employed to confirm the taxonomic provenance of strain M-43T within the genus Myroides. The overall genome relatedness indices (OGRI) for strain M-43T compared with its most closely related type strains M. odoratimimus subsp. xuanwuensis DSM 27251T, M. profundi D25T, and M. odoratimimus subsp. odoratimimus ATCC BAA-634T, were calculated as 25.3%, 25.1%, and 25% for digital DNA-DNA hybridization (dDDH), 83.3%, 83.6%, and 83.4% for average nucleotide identity (ANI) analyses, respectively. The OGRI values between strain M-43T and its close neighbors confirmed that the strain represents a novel species in the genus Myroides. The DNA G + C content of the strain is 33.7%. The major fatty acids are iso-C15:0 and summed feature 9 (iso-C17:1 ω9c and/or 10-methyl C16:0). The predominant polar lipids are phosphatidylethanolamine, an amino-lipid and five unidentified lipids. The major respiratory quinone is MK-6. Chemotaxonomic and phylogenomic analyses of this isolate confirmed that the strain represents a novel species for which the name Myroides oncorhynchi sp. nov. is proposed, with M-43T as the type strain (JCM 34205T = KCTC 82265T).

Genomic insight into Chryseobacterium turcicum sp. nov. and Chryseobacterium muglaense sp. nov. isolated from farmed rainbow trout in Turkey

Four strains, designated as C-2, C-17T, C-39T and Ch-15, were isolated from farmed rainbow trout samples showing clinical signs during an investigation for a fish-health screening study. The pairwise 16S rRNA gene sequence analysis showed that strain C-17T shared the highest identity level of 98.1 % with the type strain of Chryseobacterium piscium LMG 23089T while strains C-2, C-39T and Ch-15 were closely related to Chryseobacterium balustinum DSM 16775T with an identity level of 99.3 %. A polyphasic approach involving phenotypic, chemotaxonomic and genome-based analyses was employed to determine the taxonomic provenance of the strains. The overall genome relatedness indices including dDDH and ANI analyses confirmed that strains C-2, C-17T, C-39T and Ch-15 formed two novel species within the genus Chryseobacterium. Chemotaxonomic analyses showed that strains C-17T and C-39T have typical characteristics of the genus Chryseobacterium by having phosphatidylethanolamine in their polar lipid profile, MK-6 as only isoprenoid quinone and the presence of iso-C15:0 as major fatty acid. The genome size and G + C content of the strains ranged between 4.4 and 5.0 Mb and 33.5 – 33.6 %, respectively. Comprehensive genome analyses revealed that the strains have antimicrobial resistance genes, prophages and horizontally acquired genes in addition to secondary metabolite-coding gene clusters. In conclusion, based on the polyphasic analyses conducted on the present study, strains C-17T and C-39T are representatives of two novel species within the genus Chryseobacterium, for which the names Chryseobacterium turcicum sp. nov. and Chryseobacterium muglaense sp. nov. with the type strains C-17T (=JCM 34190T = KCTC 82250T) and C-39T (=JCM 34191T = KCTC 822251T), respectively, are proposed.

Resolution of housekeeping gene sequences used in MLSA for the genus Streptomyces and reclassification of Streptomyces anthocyanicus and Streptomyces tricolor as heterotypic synonyms of Streptomyces violaceoruber.

Although 16S rRNA gene sequences are conventionally analysed in bacterial systematics, their resolution is not sufficient for species identification. Multilocus sequence analysis (MLSA) is a powerful method for species identification as well as the elucidation of phylogenetic relationships in the genus Streptomyces. Gene sequences of atpD, gyrB, recA, rpoB and trpB are generally used in MLSA for Streptomyces species. This study aims to evaluate the sequence analysis of one gene instead of all five genes to be employed for species identification. The resolution of atpD gene sequences was not necessarily able to distinguish closely related species. In contrast, trpB gene sequence similarities correlated to the MLSA-based evolutionary distances, especially among closely related strains. A pairwise similarity of 97.9 % in trpB gene sequences was proposed as the threshold for species delineation based on the feasibility examined using strain pairs that shared >99.93 % pairwise 16S rRNA gene sequence similarities. Resequencing the five housekeeping genes followed by MLSA suggested that Streptomyces anthocyanicus and Streptomyces tricolor are synonyms of Streptomyces violaceoruber.

Pusillimonas faecipullorum sp. nov., isolated from the poultry manure.

A polyphasic taxonomic approach was used to characterize a novel bacterium, designated strain CC-YST705T, isolated from poultry manure sampled in Taiwan. Cells of strain CC-YST705T were aerobic, Gram-stain-negative, non-motile, non-spore-forming rods, displaying positive reactions for catalase, oxidase and β-glucosidase. Optimal growth occurred at 30°C and pH 8. Strain CC-YST705T shared the highest (> 96.0%) pair-wise 16S rRNA gene sequence similarity with Pusillimonas noertemannii (96.7%), followed by Pusillimonas caeni (96.6%), Eoetvoesia caeni (96.1%) and Paracandidimonas soli (96.0%), and formed a distinct phyletic lineage associate with the clade that accommodated Pusillimonas species. The draft genome (3.1Mb) having 57.4mol % G + C content contained genes involved in the catabolism of aromatic hydrocarbons. The orthologous average nucleotide identity (OrthoANI) values were 62.8-73.1% (n = 8), 72.6 (n = 1), 71.5% (n = 1) compared within the type strains of the genera Pusillimonas, Eoetvoesia and Paracandidimonas, respectively. The polar lipid profile consisted of phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, one unidentified aminolipid, two unidentified phospholipids and five unidentified lipids. The major polyamine was spermidine. The dominating cellular fatty acids (> 5%) included C12:0, C16:0, C17:0 cyclo, C19:0 cyclo ω8c and 2 C14:0 3OH/iso-C16:1 I. Based on its distinct phylogenetic, phenotypic and chemotaxonomic traits together with results of comparative 16S rRNA gene sequence, OrthoANI, digital DDH, and the phylogenomic placement, strain CC-YST705T is considered to represent a novel species of the genus Pusillimonas, for which the name Pusillimonas faecipullorum sp. nov. is proposed. The type strain is CC-YST705T (= BCRC 81285T = JCM 34168T).

Halomonas populi sp. nov. isolated from Populus euphratica.

Three yellow-pigmented, Gram-stain-negative, aerobic, motile by flagella and rod-shaped strains, designated as MCT, PC and RC, were isolated from stems of Populus euphratica. Growth of those three strains occurs at 4-40°C, pH 6.0-10.0 and with 0.5-18.0% (w/v) NaCl. Respiratory quinones contained ubiquinone-9 and ubiquione-8 as major and minor components, respectively. Major fatty acids (> 10%) were summed feature 8 (C18:1ω6c and/or C18:1ω7c), summed feature 3 (C16:1ω6c and/or C16:1ω7c) and C16:0. Polar lipids included diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, two unidentified phospholipids, one unidentified aminolipid, one unidentified glycolipid and four unidentified lipids. Strains MCT, PC and RC shared pairwise 16S rRNA gene sequence similarities of 99.9-100.0%, and showed higher similarities of 98.4-98.5% with Halomonas songnenensis NEAU-ST10-39T and 98.3-98.4% with Halomonas nanhaiensis YIM M 13059T than to other Halomonas type strains. Genomic comparisons revealed that those three strains had the pan-genome consisting of 4446 orthologous clusters, among which 676 orthologous clusters were absent in other Halomonas type strains. Phylogenomic tree indicated that strains MCT, PC and RC formed an independently stable clade with Halomonas nanhaiensis YIM M 13059T and Halomonas songnenensis NEAU-ST10-39T. The average nucleotide identity and digital DNA-DNA hybridization values between those three strains and other Halomonas type strains were < 89.9% and < 39.3%, respectively. Based upon phenotypic, chemotaxonomic, phylogenetic and genomic results, strains MCT, PC and RC represent a novel species in the genus Halomonas, for which the name Halomonas populi sp. nov. is proposed. The type strain is MCT (= JCM 33545T = MCCC 1K03942T).

Characterization of the Aerobic Anoxygenic Phototrophic Bacterium Sphingomonas sp. AAP5.

An aerobic, yellow-pigmented, bacteriochlorophyll a-producing strain, designated AAP5 (=DSM 111157=CCUG 74776), was isolated from the alpine lake Gossenköllesee located in the Tyrolean Alps, Austria. Here, we report its description and polyphasic characterization. Phylogenetic analysis of the 16S rRNA gene showed that strain AAP5 belongs to the bacterial genus Sphingomonas and has the highest pairwise 16S rRNA gene sequence similarity with Sphingomonas glacialis (98.3%), Sphingomonas psychrolutea (96.8%), and Sphingomonas melonis (96.5%). Its genomic DNA G + C content is 65.9%. Further, in silico DNA-DNA hybridization and calculation of the average nucleotide identity speaks for the close phylogenetic relationship of AAP5 and Sphingomonas glacialis. The high percentage (76.2%) of shared orthologous gene clusters between strain AAP5 and Sphingomonas paucimobilis NCTC 11030T, the type species of the genus, supports the classification of the two strains into the same genus. Strain AAP5 was found to contain C18:1ω7c (64.6%) as a predominant fatty acid (>10%) and the polar lipid profile contained phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine, sphingoglycolipid, six unidentified glycolipids, one unidentified phospholipid, and two unidentified lipids. The main respiratory quinone was ubiquinone-10. Strain AAP5 is a facultative photoheterotroph containing type-2 photosynthetic reaction centers and, in addition, contains a xathorhodopsin gene. No CO2-fixation pathways were found.

Frankia soli sp. nov., an actinobacterium isolated from soil beneath Ceanothus jepsonii.

Actinobacterial strain CjT was directly isolated from soil beneath Ceanothus jepsonii growing in the USA. The strain formed cell structures typical of the genus Frankia including extensive hyphae, vesicles and sporangia, and it effectively nodulated members of the actinorhizal Colletieae, Elaeagnaceae and Myricaceae. The whole-cell hydrolysate of strain CjT was rich in meso-diaminopimelic acid and galactose, glucose, mannose, xylose, ribose and a trace of rhamnose. Tbe polar lipid profile contained phosphatidylinositol, phosphatidylglycerol, diphosphatidylglycerol and glycophospholipid. The menaquinone was predominantly MK-9(H4). The fatty acid profile predominantly consisted of C17 : 1ω8c, iso-C16 : 0, C15:0, C16 : 0 and C17 : 0. A multilocus sequence analysis phylogeny based on atp1, ftsZ, dnaK, gyrA and secA gene sequences positioned the strain within Elaeagnaceae- and Colletieae-nodulating species together with Frankia elaeagni DSM 46783T, Frankia discariae DSM 46785T and Frankia irregularis DSM 45899T. Pairwise 16S rRNA gene sequence similarities showed that strain CjT was most closely related to F. discariae DSM 46785T (99.78 %) while their digital DNA-DNA hybridization value was 41.1 %. Based on the overall analyses, strain CjT (=DSM 100623T=CECT 9041T) warrants classification as the type strain of a novel species, for which the name Frankia soli sp. nov. is proposed.

Micromonospora radicis sp. nov., isolated from roots of Azadirachta indica var. siamensis Valenton, and reclassification of Jishengella zingiberis as Micromonospora zingiberis comb. nov.

A novel endophytic actinomycete strain AZ1-13T was isolated from roots of Azadirachta indica, and its taxonomic position was investigated using a polyphasic approach. Pairwise 16S rRNA gene sequence similarities of strain AZ1-13T and its closest species, Jishegella zingiberis PLAI1-1T and Micromonospora endophytica 202201T, were 99.7 and 99.2 %, respectively. Phylogenetic analyses of the family Micromonosporaceae based on 16S rRNA gene sequences indicated strains AZ1-13T and J. zingiberis PLAI1-1Tare located within the genus Micromonospora. The approximate genome size of the strain was 5.96 Mb with 71.9 mol% of G+C content. The strain AZ1-13T exhibited ANIb values of 87.4 % with J. zingiberis PLAI1-1T and 85.1 % with M. endophytica 202201T. Chemotaxonomic characteristics of strain AZ1-13T were consistent within the genus Micromonospora: cell-wall peptidoglycan of the strain contained meso-diaminopimelic acid; glucose, mannose, ribose and xylose are presented as the whole-cell sugars; the predominant menaquinones were MK-9(H4) and MK-9(H6); major cellular fatty acids were iso-C15 : 0, 10-methyl C17 : 0, C17 : 0, anteiso-C17 : 0 and iso-C17 : 1ω8c; diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylinositol were detected as distinguished phospholipids. Based on phenotypic properties, phylogeny and genomic data, the strain AZ1-13T could be distinguished from its closest neighbours, representing a novel species of the genus Micromonospora, for which the name Micromonospora radicis sp. nov. is proposed. The type strain is AZ1-13T (=KCTC 39786T=NBRC 112324T=JCM 32147T = TISTR 2404T). This study also proposed that J. zingiberisis transferred to the genus Micromonospora as Micromonospora zingiberis comb. nov. (type strain PLAI1-1T=TBRC 7644T=NBRC 113144T=JCM 32592T).

Mesorhizobium composti sp. nov., isolated from compost

A polyphasic taxonomic approach was used to characterize a presumptively novel diazotrophic bacterium, designated strain CC-YTH430T, isolated from a compost sample in Taiwan. Cells of strain CC-YTH430T were found to be Gram-stain negative, facultative anaerobic rods that formed yellow-colored colonies on nutrient agar. Cell growth occurred at 15-40°C, pH 5.0-9.0 and in the presence of 0-2% NaCl. Strain CC-YTH430T resembled Mesorhizobium species while sharing high pair-wise 16S rRNA gene sequence similarities with Mesorhizobium silamurunense, Mesorhizobium thiogangeticum, Mesorhizobium plurifarium, Mesorhizobium tamadayense, Mesorhizobium amorphae (96.9% each), Mesorhizobium sediminum (96.8%), and Mesorhizobium soli (96.5%) and < 96.5% similarity to other species. Strain CC-YTH430T showed 78.8-79.7% average nucleotide identity compared to the type strains of M. amorphae, M. plurifarium, M. soli, M. tamadayense and M. wenxiniae. The N2-fixing activity of strain CC-YTH430T was 0.2nmol ethyleneh-1 at 30°C. The respiratory system was ubiquinone 10 (Q-10) and the DNA G+C content was 62.0 ± 0.2mol%. The major fatty acids (> 5%) were C16:0, C17:0 cyclo, C19:0 cyclo ω8c, C14:0 3OH/C16:1 iso I and C18:1ω7c/C18:1ω6c. The polar lipid profile contained diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine, phosphatidylmonomethylethanolamine and an unidentified aminolipid in major amounts. In addition, phosphatidylethanolamine, an unidentified lipid and several unidentified polar lipids were also found in moderate-to-trace amounts. Based on the phylogenetic, phenotypic and chemotaxonomic features, strain CC-YTH430T is proposed to represent a novel Mesorhizobium species, for which the name Mesorhizobium composti sp. nov. (type strain CC-YTH430T = BCRC 81024T = JCM 31762T) is proposed.

Halioglobus sediminis sp. nov., isolated from coastal sediment.

A novel aerobic, Gram-stain-negative, non-motile and rod-shaped bacterium, designated strain U0301T, was isolated from a marine sediment sample in Weihai, China. This strain grew optimally at 33 °C, pH 7.5 and in the presence of 2.0-3.0 % (w/v) NaCl. Strain U0301T shared pairwise 16S rRNA gene sequence similarities of 95.5, 95.4, 95.2 and 95.0 % to Parahaliea aestuarii S2-26T, Halioglobus pacificus KCTC 23430T, Halioglobus lutimaris HF004T and Halioglobus japonicus KCTC 23429T, respectively. Phylogenetic analysis based on the 16S rRNA gene sequences demonstrated that U0301T formed a tight phylogenetic lineage with type strains of H. pacificus and H. japonicus. The percentage of conserved protein values of strain U0301T as compared with H. japonicus KCTC 23429Tand Parahalieamediterranea 7SM29T were 62.5 and 58.0 %, respectively. The ANI values of strain U0301T with H. japonicus KCTC 23429T and P. mediterranea 7SM29T were 78.2 and 75.3 %, respectively. Both metrics of genome comparison suggested that strain U0301T showed higher homology with the genus Halioglobus than the genus Parahaliea. The strain contained ubiquinone 8 as the sole respiratory quinone. The major fatty acids were summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c), summed feature 3 (C16 : 1ω6c and/or C16 : 1ω7c) and C17 : 1ω8c. The major polar lipids were phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol. The DNA G+C content was 61.7 mol%. Considering the phenotypic characteristics, chemotaxonomic data and phylogenetic analysis comprehensively, strain U0301T should represent a novel species of the genus Halioglobus, for which the name of Halioglobus sediminis sp. nov. is proposed. The type strain is U0301T (=KCTC 62082T=MCCC 1H00234T).

Flavobacterium petrolei sp. nov., a novel psychrophilic, diesel-degrading bacterium isolated from oil-contaminated Arctic soil

This study presents taxonomic description of two novel diesel-degrading, psychrophilic strains: Kopri-42T and Kopri-43, isolated during screening of oil-degrading psychrotrophs from oil-contaminated Arctic soil. A preliminary 16S rRNA gene sequence and phylogenetic tree analysis indicated that these Arctic strains belonged to the genus Flavobacterium, with the nearest relative being Flavobacterium psychrolimnae LMG 22018T (98.9% sequence similarity). The pairwise 16S rRNA gene sequence identity between strains Kopri-42T and Kopri-43 was 99.7%. The DNA-DNA hybridization value between strain Kopri-42T and Kopri-43 was 88.6 ± 2.1% indicating that Kopri-42T and Kopri-43 represents two strains of the same genomospecies. The average nucleotide identity and in silico DNA-DNA hybridization values between strain Kopri-42T and nearest relative F. psychrolimnae LMG 22018T were 92.4% and 47.9%, respectively. These values support the authenticity of the novel species and confirmed the strain Kopri-42T belonged to the genus Flavobacterium as a new member. The morphological, physiological, biochemical and chemotaxonomic data also distinguished strain Kopri-42T from its closest phylogenetic neighbors. Based on the polyphasic data, strains Kopri-42T and Kopri-43 represents a single novel species of the genus Flavobacterium, for which the name Flavobacterium petrolei sp. nov. is proposed. The type strain is Kopri-42T (=KEMB 9005-710T = KACC 19625T = NBRC 113374T).

Streptacidiphilus bronchialis sp. nov., a ciprofloxacin-resistant bacterium from a human clinical specimen; reclassification of Streptomyces griseoplanus as Streptacidiphilus griseoplanus comb. nov. and emended description of the genus Streptacidiphilus.

The taxonomic position of strain 15-057AT, an acidophilic actinobacterium isolated from the bronchial lavage of an 80-year-old male, was determined using a polyphasic approach incorporating morphological, phenotypic, chemotaxonomic and genomic analyses. Pairwise 16S rRNA gene sequence similarities calculated using the GGDC web server between strain 15-057AT and its closest phylogenetic neighbours, Streptomyces griseoplanus NBRC 12779T and Streptacidiphilus oryzae TH49T, were 99.7 and 97.6 %, respectively. The G+C content of isolate 15-057AT was determined to be 72.6 mol%. DNA-DNA relatedness and average nucleotide identity between isolate 15-057AT and Streptomyces griseoplanus DSM 40009T were 29.2±2.5 % and 85.97 %, respectively. Chemotaxonomic features of isolate 15-057AT were consistent with its assignment within the genus Streptacidiphilus: the whole-cell hydrolysate contained ll-diaminopimelic acid as the diagnostic diamino acid and glucose, mannose and ribose as cell-wall sugars; the major menaquinone was MK9(H8); the polar lipid profile consisted of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, glycophospholipid, aminoglycophospholipid and an unknown lipid; the major fatty acids were anteiso-C15 : 0 and iso-C16 : 0. Phenotypic and morphological traits distinguished isolate 15-057AT from its closest phylogenetic neighbours. The results of our taxonomic analyses showed that strain 15-057AT represents a novel species within the evolutionary radiation of the genus Streptacidiphilus, for which the name Streptacidiphilus bronchialis sp. nov. is proposed. The type strain is 15-057AT (=DSM 106435T=ATCC BAA-2934T).

Nocardioides phosphati sp. nov., an actinomycete isolated from a phosphate mine.

A Gram-staining-positive, non-spore-forming and non-motile strain, designated WYH11-7T, was isolated from a phosphate mine in Yunnan Province, PR China. The taxonomic position of WYH11-7T was investigated by polyphasic approaches. Phylogenetic analyses based on 16S rRNA gene sequences indicated that WYH11-7T represents a member of the genus Nocardioides. WYH11-7T was closely related to Nocardioidesjensenii DSM 20641T, Nocardioidesdubius DSM 19084T and Marmoricolaterrae DSM 27141T, and had pairwise 16S rRNA gene sequence similarities of 97.4, 97.2 and 97.0 %, respectively. DNA-DNA relatedness values between WYH11-7T and related type strains N. jensenii DSM 20641T and N. dubius DSM 19084T were found to be 17.6±4.9 and 14.6±3.1 %, respectively. The respiratory menaquinone of WYH11-7T was MK-8 (H4) while the major fatty acids were C18 : 1ω9c, C16 : 0, C17 : 0, C17 : 1ω8c, C18 : 1 10-methyl and summed feature 3 (C16 : 1ω6c and/or C16 : 1ω7c). The polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol and two unidentified phospholipids. Whole-cell hydrolysates contained mannose, ribose, glucose and galactose along with ll-diaminopimelic acid as the diagnostic diamino acid in the peptidoglycan. The DNA G+C content was 71.2 mol%. Phenotypic, phylogenetic and chemotaxonomic data indicated that strain WYH11-7T represents a novel species of the genus Nocardioides, for which the name Nocardioidesphosphatisp. nov. is proposed. The type strain is WYH11-7T (=CGMCC 4.7371T=DSM 104026T).

Ruficoccus amylovorans gen. nov., sp. nov., an amylolytic and nitrate-reducing diazotroph of the family Puniceicoccaceae.

A novel amylolytic, nitrate-reducing and diazotrophic bacterium, designated strain CC-MHH0563T, isolated from a fermenter was assessed for its taxonomic status using a polyphasic approach. Cells of strain CC-MHH0563T were Gram-staining-negative, catalase- and oxidase-positive, mesophilic and aerobic cocci, which produced reddish nondiffusible pigments. Growth was observed at 15-37 °C (optimal 25 °C), pH 6.0-8.0 (optimal pH 7.0) and salinity of 0-3 % (w/v). Strain CC-MHH0563T showed highest pairwise 16S rRNA gene sequence similarity to members of the genera Cerasicoccus(89.3-89.5 %), Coraliomargarita (87.8 %), Pelagicoccus(85.8-86.4 %) and Puniceicoccus (87.9 %), and established a discrete taxonomic lineage during phylogenetic analysis. The major fatty acids found in strain CC-MHH0563T were C14 : 0, anteiso-C15 : 0, C16 : 0, C17 : 0, C18 : 0 and C18 : 1ω9c. The polar lipid profile consisted of phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, two unknown aminolipids and two unknown aminophospholipids. The polyamine pattern showed a predominance of spermidine and a minor amount of cadaverine. The DNA G+C content was 57.4 mol% and the predominant quinone system was menaquinone-7. The low 16S rRNA gene sequence similarity values (<90.0 %) and a distinct phylogenetic clustering clearly distinguished strain CC-MHH0563T from other representatives of the family Puniceicoccaceae. Based on the discrete phylogenetic, phenotypic and chemotaxonomic traits together with the results of comparative 16S rRNA gene sequence analysis, strain CC-MHH0563T is considered to represent a novel genus and species of the family Puniceicoccaceae, for which the name Ruficoccus amylovorans gen. nov., sp. nov. is proposed. The type strain of the type species is CC-MHH0563T (=BCRC 80918T=JCM 31066T).

Sphingobacterium cibi sp. nov., isolated from the food-waste compost and emended descriptions of Sphingobacterium spiritivorum (Holmes et al. 1982) Yabuuchi et al. 1983 and Sphingobacteriumthermophilum Yabe et al. 2013.

A novel, pale yellow-coloured, Gram-staining-negative, aerobic, rod-shaped and non-motile bacterium, designated as strain CC-YY411T, was isolated from the food-waste compost and subjected to polyphasic taxonomy. Strain CC-YY411T exhibited the highest pairwise 16S rRNA gene sequence similarity to Sphingobacterium thermophilum JCM 17858T (94.6 %) while sharing 94.1 to 89.7 % similarities with other Sphingobacterium species. Novel strain established a discrete phylogenetic lineage within the clade that accommodated validly established members of the genus Sphingobacterium. The polar lipid profile of strain CC-YY411T contained major amounts of phosphatidylethanolamine, one unidentified lipid and two unidentified aminolipids besides accommodating trace amounts of a sphingolipid, two phospholipids, an unidentified aminophospholipid and an unidentified glycolipid. The DNA G+C content of strain CC-YY411T was 34.5 mol%. The major and minor respiratory quinones were MK-7 (89.1 %) and MK-6 (10.9 %), respectively. The predominant fatty acids were iso-C15 : 0 (24.0 %), iso-C17 : 0 3-OH (11.9 %), iso-C15 : 0 3-OH (6.9 %), C15 : 1ω5c (5.5 %) and summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c) (32.0 %). Based on the phylogenetic, phenotypic and chemotaxonomic distinctiveness, strain CC-YY411T represents a novel species of Sphingobacterium, for which the name Sphingobacteriumcibi sp. nov. is proposed. The type strain is CC-YY411T (=BCRC 80430T=JCM 18407T). Amended species descriptions of Sphingobacterium spiritivorum(Holmes et al. 1982) Yabuuchi et al. 1983 and Sphingobacterium thermophilum Yabe et al. 2013 are also proposed based on new data obtained in this study.

Nocardioides gilvus sp. nov., isolated from Namtso Lake.

A Gram-stain positive, facultatively anaerobic, non-motile and rod to coccoid-shaped bacterium, designated XZ17(T), was isolated from Namtso Lake of Tibet, China. Strain XZ17(T) grew optimally at pH 8.0-9.0, at 30-33°C and in the presence of 0-1.0% (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences demonstrated that strain XZ17(T) belongs to the genus Nocardioides in the family Nocardioidaceae. Strain XZ17(T) shared pairwise 16S rRNA gene sequence similarities of 97.2, 96.8, 96.5, 96.4 and <96.0% to Nocardioides solisilvae KCTC 39528(T), Nocardioides daejeonensis JCM 16922(T), Nocardioides jensenii NCIB 9770(T), Nocardioides dubius KCTC 9992(T) and other Nocardioides species, respectively. It contained MK-8 (H4) as the predominant menaquinone and C17:1 ω8c, C15:0, C17:0 and C18:1 ω9c as the major fatty acid. The strain had cell wall peptidoglycan based on LL-2,6-diaminopimelic acid. The polar lipids of strain XZ17(T) comprised diphosphatidylglycerol, phosphatidylglycerol, three unidentified phospholipids, three unidentified glycolipids and two unidentified lipids. The DNA G+C content of strain XZ17(T) was 68.9mol%. Based on distinctive phenotypic characteristics, phylogenetic analysis and chemotaxonomic data, it can be concluded that strain XZ17(T) represents a novel species within the genus Nocardioides, for which the name Nocardioides gilvus sp. nov. is proposed. The type strain is strain XZ17(T) (=KCTC 39561(T)=MCCC 1H00114(T)).

Nocardioides intraradicalis sp. nov., isolated from the roots of Psammosilene tunicoides W. C. Wu et C. Y. Wu.

A Gram-stain-positive, non-spore-forming and non-motile strain, designated YIM DR1091T, was isolated from the roots of Psammosilene tunicoides W. C. Wu et C. Y. Wu collected from Gejiu, Yunnan, China. The taxonomic position of strain YIM DR1091T was investigated by a polyphasic approach. Phylogenetic analyses based on 16S rRNA gene sequences indicated that strain YIM DR1091T is a member of the genus Nocardioides. Strain YIM DR1091T was closely related to Nocardioides pyridinolyticus OS4T, Nocardioides caricicola YC6903T, Nocardioides hankookensis DS-30T and Nocardioides aquiterrae GW-9T, with which it shared pairwise 16S rRNA gene sequence similarities of 97.6, 97.5, 97.2 and 97.2 %, respectively. Mean DNA-DNA relatedness values between strain YIM DR1091T and related type strains N. pyridinolyticus JCM 10369T, N. caricicola JCM 17686T, N. hankookensis JCM 15302T and N. aquiterrae JCM 11813T were 44.9±1.7, 50.2±1.3, 46.8±0.9 and 43.0±0.2 %, respectively. The respiratory menaquinone for strain YIM DR1091T was MK-8(H4) while the major fatty acids (>5 %) were iso-C16 : 0, C17 : 1ω8c, C17 : 0, iso-C15 : 0 and iso-C14 : 0. The polar lipids were diphosphatidylglycerol, phosphatidylglycerol and three unidentified phospholipids. Whole-cell hydrolysates contained mannose, ribose, glucose and galactose, along with ll-diaminopimelic acid as the diagnostic diamino acid in the peptidoglycan. The DNA G+C content was 74.6 mol%. Phenotypic, phylogenetic and chemotaxonomic data indicated that strain YIM DR1091T represents a novel species of the genus Nocardioides, for which the name Nocardioides intraradicalis sp. nov. is proposed. The type strain is YIM DR1091T (=JCM 30632T=CGMCC4.7251T).

Streptomyces zhihengii sp. nov., isolated from rhizospheric soil of Psammosilene tunicoides.

An actinomycete strain, designated YIM T102(T), was isolated from the rhizospheric soil of Psammosilene tunicoides W. C. Wu et C. Y. Wu collected from Lijiang, YunnanProvince, China. The taxonomic position of the new isolate was investigated by a polyphasic approach. Phylogenetic analyses based on 16S rRNA gene sequences indicated that strain YIM T102(T) belongs to the genus Streptomyces. Strain YIM T102(T) was most closely related to Streptomyces eurocidicus NRRL B-1676(T) with a pairwise 16S rRNA gene sequence similarity of 98.9%. However, DNA-DNA relatedness value between strain YIM T102(T) and S. eurocidicus NBRC 13491(T) was found to be 37.8±1.8%. The menaquinone composition detected for strain YIM T102(T) was MK-9 (H6) and MK-9 (H8), while the major fatty acids were summed feature 4 (38.0%), anteiso-C15:0 (13.1%), iso-C16:0 (10.1%), summed feature 3 (9.8%) and C16:0 (9.0%) and iso-C15:0 (5.2%). The whole-cell hydrolysates contained galactose, glucose, ribose and mannose, along with LL-diaminopimelic acid as the diagnostic diamino acid in the peptidoglycan. The DNA G+C content was 70.7mol%. Strain YIM T102(T) also exhibited antagonistic activity against Alternaria alternata, Alternaria brassicae and Colletotrichum nicotianae Averna, based on the findings from the comparative analyses of phenotypic and genotypic characteristics; it is proposed that strain YIM T102 represents a novel species of the genus Streptomyces, for which the name Streptomyces zhihengii sp. nov. is proposed. The type strain is YIM T102(T) (=KCTC 39115(T)=DSM 42176(T)=CGMCC 4.7248(T)).