86 Characterization and differentiation of binding modes of water-soluble porphyrins at complexation with DNA

The influence of water-soluble cationic meso-tetra-(4?N-oxyethylpyridyl)porphyrin (H₂TOEPyP4) and it’s metallocomplexes with Ni, Cu, Co, and Zn on hydrodynamic and spectral behavior of DNA solutions has been studied by UV/Vis absorption and viscosity measurement. It was shown that the presence of planar porphyrins such as H₂TOEPyP4, NiTOEPyP4, and СuTOEPyP4 leads to an increase in viscosity at relatively small concentrations, and then decrease to stable values. Such behavior is explained by intercalation of these porphyrins in DNA structure because the intercalation mode involves the insertion of a planar molecule between DNA base pairs which results in a decrease in the DNA helical twist and lengthening of the DNA. Further decrease of viscosity is explained by the saturation intercalation sites and occurs outside the binding mode. But, in the case of porphyrins with axial ligands such as CoTOEPyP4 and ZnTOEPyP4, the hydrodynamic parameters decrease, which is explained by self-stacking of these porphyrins in DNA surface. This data are proved by spectral measurements. The results obtained from titration experiments were used for calculation of binding parameters: the binding constant K _b and the number of binding sites per base pair n. Obtained data reveal that K _b varies between 3.4 and 5.4?×?10⁶?M^?1 for a planar porphyrins, a range typical for intercalation mode interactions, and 5.6?×?10⁵?M^?1 and 1.8?×?10⁶?M^?1 for axial porphyrins. In addition, the exclusion parameter n also testifies that at intercalation, (n～2) the adjacent base pairs are removed to place the planar molecules, and for outside binders to pack on the surface needs too few places (n～0.5–1). It is apparent that the binding is somewhat stronger at intercalation. The viscometric and spectrophotometric measurements are in good agreement.     

<Emphasis Type="Italic">Baekduia soli</Emphasis> gen. nov., sp. nov., a novel bacterium isolated from the soil of Baekdu Mountain and proposal of a novel family name, <Emphasis Type="Italic">Baekduiaceae</Emphasis> fam. nov.

A taxonomic study was conducted on BR7-21^T, a bacterial strain isolated from the soil of a ginseng field in Baekdu Mountain. Comparative studies of the 16S rRNA gene sequence showed that the isolate was most closely related to Conexibacter woesei DSM 14684^T, Solirubrobacter pauli ATCC BAA-492^T, Patulibacter minatonensis JCM 12834^T, with 93.8%, 92.4%, and 91.5% sequence similarity, respectively; each genus represented a family in the order Solirubrobacterales. Strain BR7-21^T was Gram-reaction positive, non-spore forming, aerobic, non-motile, and short rod-shaped. It grew well on half-strength R2A medium. The G + C content of the genomic DNA was 73.9%. It contained meso-diaminopimelic acid in the cell wall and the major menaquinones were MK-7(H₄) and MK-8(H₄). The major fatty acids were summarized as (C_16:1ω7c/iso-C_15:0 2-OH), iso-C_16:0, and C_17:0 cyclo. On the basis of polyphasic evidence, it was proposed that strain BR7-21^T should be placed in a new genus and species, for which the name Baekduia soli gen. nov., sp. nov. was proposed with the type strain BR7-21^T (= KCTC 22257T = LMG 24797^T). The family Baekduiaceae fam. nov. is proposed to encompass the genus Baekduia gen. nov.     

<Emphasis Type="Italic">Streptomyces carminius</Emphasis> sp. nov., a novel actinomycete isolated from <Emphasis Type="Italic">Sophora alopecuroides</Emphasis> in Xinjiang,China

A novel actinobacterial strain, designated TRM SA0054^T, was isolated from the roots of Sophora alopecuroides grown in Alar, Xinjiang, north-west China, and characterised by a polyphasic taxonomic approach. Phylogenetic analysis showed that strain TRM SA0054^T has 16S rRNA gene sequence similarity of 98.22% with Streptomyces barkulensis RC 1831^T. Whole cell hydrolysates of strain TRM SA0054^T were found to contain ll-diaminopimelic acid as the diagnostic diamino acid and ribose, glucose and xylose as the major whole cell sugars. The major fatty acids were identified as iso-C_16:0, iso-C_16:1 G, anteiso-C_17:0, anteiso-C_15:0, C_16:0, anteiso-C_17:1 ω9c. The main menaquinones were determined to be MK-8 (H₄), MK-9 (H₆) and MK-9 (H₈). The polar lipids were identified as diphosphatidylglycerol, phosphatidylmethylethanolamine, phosphatidylethanolamine, phosphatidylcholine, phosphatidylinositol, phosphatidylinositol mannoside and an unidentified lipid. The G?+?C content of the genomic DNA was determined to be 73.04%. Strain TRM SA0054^T has a relatively low DNA–DNA relatedness value with Streptomyces barkulensis RC 1831^T as determined by calculating the average nucleotide identity value (ANI?=?84.1%). Based on the phenotypic, chemotaxonomic and phylogenetic data, it is concluded that strain TRM SA0054^T should be designated as a novel species of the genus Streptomyces, for which the name Streptomyces carminius sp. nov. is proposed, with type strain TRM SA0054^T (=?CCTCC AA 2016041^T?=?KCTC39903^T).     

<Emphasis Type="Italic">Flavobacterium knui</Emphasis> sp. nov., a novel member of the family <Emphasis Type="Italic">Flavobacteriaceae</Emphasis>

A Gram-stain negative, non-motile, rod-shaped bacterial strain, designated 2-56^T, was isolated from water and characterized taxonomically using a polyphasic approach. Comparative 16S rRNA gene sequence analysis showed that strain 2-56^T belongs to the family Flavobacteriaceae in the phylum Bacteroidetes and is closely related to Flavobacterium paronense KNUS1^T (98.4%) and Flavobacterium collinsense 4-T-2^T (96.7%). The G?+?C content of the genomic DNA of strain 2-56^T was 33.4 mol%. The isolate contained MK-6 as the predominant respiratory quinone, and iso-C_15:1 G (15.9%), iso-C_15:0 (15.8%), iso-C_17:0 3-OH (10.7%), and iso-C_15:0 3-OH (9.6%) were the major fatty acids. The major polar lipids were phosphatidylethanolamine and an unidentified lipid. The phenotypic and chemotaxonomic data support the affiliation of strain 2-56^T with the genus Flavobacterium. However, the DNA–DNA relatedness between the isolate and F. paronense and F. collinsense were 35.7 and 21.5%, respectively, clearly showing that strain 2-56^T is not related to them at the species level. Strain 2-56^T could be clearly differentiated from its close neighbours on the basis of its phenotypic, genotypic and chemotaxonomic features. Therefore, strain 2-56^T represents a novel species of the genus Flavobacterium, for which the name Flavobacterium knui sp. nov. is proposed. The type strain is 2-56^T (=?KCTC 62061^T?=?JCM 32247^T).     

<Emphasis Type="Italic">Planomonospora algeriensis</Emphasis> sp. nov., an actinobacterium isolated from a Saharan soil of Algeria

A filamentous actinobacterium, designated strain PM3^T, was isolated from a Saharan soil sample collected from Béni-Abbès, Béchar (South-West Algeria). A polyphasic taxonomic study was carried out to establish the status of strain PM3^T. The isolate was found to have morphological and chemotaxonomical properties associated with members of the genus Planomonospora. The new isolated microorganism developed cylindrical sporangia arranged in double parallel rows on aerial mycelium, each one containing a motile single sporangiospore. The cell wall of the strain was found to contain meso-diaminopimelic acid. Whole-cell hydrolysates were found to contain madurose, glucose, mannose and ribose. The predominant menaquinone was identified as MK-9(H₂) (69.6%). The polar lipids detected were identified as diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, phosphatidylethanolamine, phosphatidylhydroxyethanolamine and glucosamine-containing lipids. The major fatty acids were found to be C_17:1ω9c (38.6%) and C_17:0 (24.2%). Results of 16S rRNA gene sequence comparison revealed that strain PM3^T shared a high degree of 16S rRNA gene sequence similarity with Planomonospora sphaerica DSM 44632^T (99.3%), Planomonospora parontospora subsp. parontospora DSM 43177^T (99.2%) and P. parontospora subsp. antibiotica DSM 43869^T (99.0%). DNA–DNA hybridization values between strain PM3^T and the type strains of the closely related species were between 58.4 and 70.1%. The combination of phylogenetic analysis, DNA–DNA relatedness data, phenotypic characteristics and chemotaxonomic data support the conclusion that strain PM3^T represents a novel species of the genus Planomonospora, for which the name Planomonospora algeriensis sp. nov. is proposed. The type strain is PM3^T (=DSM 46752^T = CECT 9047^T).     

<Emphasis Type="Italic">Flavobacterium zaozhuangense</Emphasis> sp. nov., a new member of the family <Emphasis Type="Italic">Flavobacteriaceae</Emphasis>, isolated from metolachlor-contaminated soil

Strain ZZ-8^T, a Gram-negative, aerobic, non-spore-forming, non-motile, yellow-pigmented, rod-shaped bacterium, was isolated from metolachlor-contaminated soil in China. The taxonomic position was investigated using a polyphasic approach. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain ZZ-8^T is a member of the genus Flavobacterium and shows high sequence similarity to Flavobacterium humicola UCM-46^T (97.2%) and Flavobacterium pedocola UCM-R36^T (97.1%), and lower (<?97%) sequence similarity to other known Flavobacterium species. Chemotaxonomic analysis revealed that strain ZZ-8^T possessed MK-6 as the major respiratory quinone; and iso-C_15:0 (28.5%), summed feature 9 (iso-C_17:1 w9c/C_16:0 10-methyl, 22.9%), iso-C_17:0 3-OH (17.0%), iso-C_15:0 3-OH (8.9%), iso-C_15:1 G (8.6%) and summed feature 3 (C_16:1 w7c/C_16:1 w6c, 5.7%) as the predominant fatty acids. The polar lipids of strain ZZ-8^T were determined to be lipids, a glycolipid, aminolipids and phosphatidylethanolamine. Strain ZZ-8^T showed low DNA–DNA relatedness with F. pedocola UCM-R36^T (43.23?±?4.1%) and F. humicola UCM-46^T (29.17?±?3.8%). The DNA G+C content was 43.3 mol%. Based on the phylogenetic and phenotypic characteristics, chemotaxonomic data and DNA–DNA hybridization, strain ZZ-8^T is considered a novel species of the genus Flavobacterium, for which the name Flavobacterium zaozhuangense sp. nov. (type strain ZZ-8^T?=?KCTC 62315 ^T?=?CCTCC AB 2017243^T) is proposed.     

<Emphasis Type="Italic">Roseovarius tibetensis</Emphasis> sp. nov., a halophilic bacterium isolated from Lake LongmuCo on Tibetan Plateau

Two Gram-stain negative halophilic strains, designated as LM2^T and LM4, were isolated from Lake LongmuCo on Tibetan Plateau. These two strains were aerobic, catalaseand oxidase-positive, nonmotile and rod-shaped organisms. Phylogenetic analysis based on 16S rRNA gene sequences indicated that LM2^T and LM4 belong to the genus Roseovarius, with Roseovarius tolerans EL-172^T (97.3% and 97.4% 16S rRNA gene sequence similarity, respectively) and Roseovarius azorensis SSW084^T (95.5% and 95.6% 16S rRNA gene sequence similarity, respectively) as their closest neighbors. Q-10 was the sole respiratory quinone of these two strains. The major fatty acids were C_18:1ω7c/C_18:1ω6c, C_16:0, C_19:0 cyclo ω8c, and 11-methyl C_18:1ω7c. The polar lipids included phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, phospholipid of unknown structure containing glucosamine, and unidentified aminolipid. The DNA G + C content was between 64.2 and 64.5 mol%. DNA-DNA hybridization showed 96.7% relatedness between LM2^T and LM4, 24.9% relatedness between LM2^T and R. tolerans EL-172^T, and 36.3% relatedness between LM4 and R. tolerans EL-172^T. Based on phylogenetic analysis, DNA-DNA hybridization, a range of physiological and biochemical characteristics, LM2^T and LM4 belong to the same species and were clearly distinguished from the type strains of the genus Roseovarius. It was evident that LM2^T and LM4 could be classified as a novel species of the genus Roseovarius, for which the name Roseovarius tibetensis sp. nov. is proposed. The type strain is LM2^T (= CGMCC 1.16230^T = KCTC 62028^T).     

<Emphasis Type="Italic">Streptomyces lutosisoli</Emphasis> sp. nov., a novel actinomycete isolated from muddy soil

A novel Gram-stain positive, spore-forming, aerobic actinomycete, designated strain NEAU-QTH3-11^T, was isolated from muddy soil collected from a stream in Qitaihe, Heilongjiang Province, northeast China and characterised using a polyphasic approach. The 16S rRNA gene sequence analysis showed that strain NEAU-QTH3-11^T belongs to the genus Streptomyces and is closely related to Streptomyces rhizosphaerihabitans NBRC 109807^T (99.38%) and Streptomyces mirabilis JCM 4791^T (99.03%). Phylogenetic analysis based on the 16S rRNA gene sequences indicated that the strain formed a cluster with S. rhizosphaerihabitans NBRC 109807^T and Streptomyces siamensis NBRC 108799^T (98.62%). The menaquinones were identified as MK-9(H₈), MK-9(H₆) and MK-9(H₄). The phospholipid profile was found to consist of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, an unidentified phospholipid and an unidentified lipid. The major fatty acids were identified as anteiso-C_15:0, iso-C_16:0, C_16:0 and C_15:0. However, multilocus sequence analysis based on five house-keeping genes (atpD, gyrB, rpoB, recA and trpB), low DNA-DNA hybridization results and some phenotypic, physiological and biochemical properties could differentiate the strain from its close relatives in the genus Streptomyces. Therefore, strain NEAU-QTH3-11^T is considered to represent a novel species of the genus Streptomyces, for which the name Streptomyces lutosisoli sp. nov. is proposed, with NEAU-QTH3-11^T (=DSM 42165^T=CGMCC 4.7198^T) as the type strain.     

<Emphasis Type="Italic">Vibrio injenensis</Emphasis> sp. nov., isolated from human clinical specimens

Vibrio species are well known as motile, mostly oxidase-positive, facultative anaerobic Gram-negative bacteria. They are abundant in aquatic environments and are a common cause of human infections including diarrhea, soft tissue diseases, and bacteremia. Here, two Gram-negative bacteria, designated M12-1144^T and M12-1181, were isolated from human clinical specimens and identified using a polyphasic taxonomic approach. Phylogenetic study based on 16S rRNA gene sequence analysis revealed that the isolates belong to the genus Vibrio, and are closely related to Vibrio metschnikovii KCTC 32284^T (98.3%) and Vibrio cincinnatiensis KCTC 2733^T (97.8%). The major fatty acids were summed feature 3 (C_16:1 ω7c/C_16:1 ω6c, 38.0%), C_16:0 (23.0%), and summed feature 8 (C_18:1 ω7c or C_18:1 ω6c, 19.3%) and major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, and phosphatidylethanolamine. The G + C content of the genomic DNA was determined to be 44.1 mol%. DNA–DNA relatedness between the two newly isolated strains and V. metschnikovii KCTC 32284^T and V. cincinnatiensis KCTC 2733^T was between 42.6 to 47.5%. The similarities of genome-to-genome distance between M12-1144^T and related species ranged from 18.4-54.8%. Based on these results, a new species of the genus Vibrio, Vibrio injenensis is proposed. The type strain is M12-1144 ^T(=KCTC 32233^T =JCM 30011^T).     

<Emphasis Type="Italic">Limnobacter humi</Emphasis> sp. nov., a thiosulfate-oxidizing,heterotrophic bacterium isolated from humus soil,and emended description of the genus <Emphasis Type="Italic">Limnobacter</Emphasis> Spring <Emphasis Type="Italic">et al.</Emphasis> 2001

Three Gram-negative, strictly aerobic, chemolithoheterotrophic bacterial strains, designated UCM-30, UCM-33, and UCM-39^T, were isolated in South Korea. Based on their 16S rRNA gene sequences, the three isolated strains were found to be similar to Limnobacter thiooxidans CS-K2^T (97.41–97.68%), Limnobacter litoralis KP1-19^T (95.55–95.76%), and various genera belonging to the class Betaproteobacteria (90.34–93.34%). DNA-DNA hybridization showed 79.3–83.9% similarity between the genomic DNA of UCM-39^T, UCM-30, and UCM-33, while the sequence similarity between UCM-39^T and L. thiooxidans KACC 13837T or L. litoralis LMG 24869T was 23.7% and 18.6%, respectively. The DNA G+C content of UCM 39T was 59.7 mol%, the major ubiquinone was Q-8, and the optimal oxidation rate was observed at 10 mM thiosulfate. The major fatty acids (≥ 10%) were summed features 3 (C_16:1 ω7c and/or C_16:1 ω6c) and 8 (C_18:1 ω7c and/or C_18:1 ω6c), and C_16:0. The major polar lipids (diphosphatidylglycerol, phosphatidylethanolamine, and phosphatidylglycerol) were found in all members of genus Limnobacter. Based on phenotypic, physiological, and phylogenetic analyses, the UCM-39T strain was found to be significantly distinct to represent a novel species affiliated to the genus Limnobacter. We propose to name it Limnobacter humi sp. nov. with the type strain UCM-39^T (=KACC 18574^T =NBRC 111650^T).     

<Emphasis Type="Italic">Paenibacillus bryophyllum</Emphasis> sp. nov., a nitrogen-fixing species isolated from <Emphasis Type="Italic">Bryophyllum pinnatum</Emphasis>

A nitrogen-fixing, endospore-forming bacterium, designated strain L201^T was isolated from the leaves of Bryophyllum pinnatum growing in South China Agricultural University. Phylogenetic analysis of the 16S rRNA gene sequence indicated that strain L201^T is affiliated with the genus Paenibacillus, and closely related to Paenibacillus albidus Q4-3^T (97.4%), Paenibacillus odorifer DSM 15391^T (97.3%) and Paenibacillus borealis DSM 13188^T (97.2%). The main fatty acids components was anteiso-C_15:0 (48.1%). The predominant isoprenoid quinone was MK-7. The major polar lipids were found to be diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylglycerol. The G+C content of strain L201^T was 43.9%. DNA–DNA relatedness between L201^T and the reference strain was 29.8%. Biological and biochemical tests, protein patterns, genomic DNA fingerprinting and comparison of cellular fatty acids distinguished strain L201^T from the closely related Paenibacillus species. Based on these data, the novel species Paenibacillus bryophyllum sp. nov. is proposed, with the type strain L201^T(=?KCTC 33951 ^T?=?GDMCC 1.1251 ^T).     

Description of <Emphasis Type="Italic">Hymenobacter daejeonensis</Emphasis> sp. nov., isolated from grass soil,based on multilocus sequence analysis of the 16S rRNA gene, <Emphasis Type="Italic">gyr</Emphasis>B and <Emphasis Type="Italic">tuf</Emphasis> genes

A polyphasic taxonomic study was carried out on strains PB105^T and PB108 isolated from a grass soil in Korea. The cells of the strains were Gram-stain negative, non-spore-forming, non-motile, and rod-shaped. Comparative 16S rRNA gene sequence studies showed a clear affiliation of these strains with Bacteroidetes, which showed high pairwise sequence similarities with Hymenobacter algoricola VUG-A23a^T (99.2%), Hymenobacter fastidiosus VUG-A124a^T (97.4%), and Hymenobacter daecheongensis Dae14^T (96.9%). The phylogenetic analysis based on 16S rRNA gene sequences showed that the strains formed a clear phylogenetic lineage with the genus Hymenobacter. The major fatty acids were identified as C_15:0 iso, C_15:0 anteiso, C_16:1 ω5c, C_15:0 iso 3-OH, C_17:0 iso 3-OH, summed feature 3 (C_16:1 ω6c and/or C_16:1 ω7c/t), and summed feature 4 (C_17:1 anteiso B and/or C_17:1 iso I). The major cellular polar lipids were identified as phosphatidylethanolamine, an unidentified aminolipid, and two unidentified lipids. The respiratory quinone was identified as MK-7 and the genomic DNA G+C content was determined to be 64.5 mol% for strain PB105^T and 64.1 mol% for strain PB108. DNA–DNA hybridization value of type strain PB105^T with H. algoricola VUG-A23a^T was 32.3% (reciprocal 39.2). Based on the combined genotypic and phenotypic data, we propose that strains PB105^T and PB108 represent a novel species of the genus Hymenobacter, for which the name Hymenobacter daejeonensis sp. nov. is proposed. The type strain is PB105^T (=?KCTC 52579^T?=?JCM 31885^T).     

<Emphasis Type="Italic">Pedobacter panacis</Emphasis> sp. nov., isolated from <Emphasis Type="Italic">Panax ginseng</Emphasis> soil

A novel strain, DCY108^T was isolated from soil of a Panax ginseng field, Yeoncheon province (38°04′N 126°57′E), Republic of Korea. Strain DCY108^T is Gram-negative, non-motile, non-flagellate, rod-shaped, and aerobic. The bacterium grows optimally at 25–30 °C, pH 6.5–7.0 and 1 % NaCl. Phylogenetically, strain DCY108^T is closely related to Pedobacter jejuensis JCM 18824^T, Pedobacter aquatilis JCM 13454^T, Pedobacter kyungheensis LMG 26577^T and the type strain of the genus Pedobacter heparinus DSM 2366^T. The DNA–DNA relatedness values between strain DCY108^T and its close phylogenetic neighbors were below 30.0 %. The DNA G+C content of strain DCY108^T was determined to be 45.1 mol%. The predominant quinone was menaquinone 7 (MK-7). The major polar lipids were identified as phosphatidylethanolamine and three unidentified aminolipids AL1, AL13 and AL17. Iso-C_15:00, iso-C_17:03OH and summed feature 3 (C_16:1 ω7c/C_16:1 ω6c) were identified as the major fatty acids present in strain DCY108^T. The results of physiological and biochemical tests allowed strain DCY108^T to be differentiated phenotypically from other recognized species belonging to the genus Pedobacter. Therefore, it is suggested that the newly isolated organism represents a novel species, for which the name Pedobacter panacis sp. nov is proposed with the type strain designated as DCY108^T (=CCTCCAB 2015196^T = KCTC 42748^T).     

<Emphasis Type="Italic">Nibribacter flagellatus</Emphasis> sp. nov., isolated from rhizosphere of <Emphasis Type="Italic">Hibiscus syriacus</Emphasis> and emended description of the genus <Emphasis Type="Italic">Nibribacter</Emphasis>

A Gram-stain negative, aerobic, motile by flagella, rod-shaped strain (THG-T16^T) 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-T16^T were identified as Nibribacter koreensis KACC 16450^T (98.6%), Rufibacter roseus KCTC 42217^T (94.7%), Rufibacter immobilis CCTCC AB 2013351^T (94.5%) and Rufibacter tibetensis CCTCC AB 208084^T (94.4%). The DNA G+C content of strain THG-T16^T was determined to be 46.7 mol%. DNA–DNA hybridization values between strain THG-T16^T and N. koreensis KACC 16450^T, R. roseus KCTC 42217^T, R. immobilis CCTCC AB 2013351^T, R.tibetensis CCTCC AB 208084^T 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 C_16:1 ω5c, C_17:1 ω6c, iso-C_15:0, summed feature 3 (C_16:1 ω7c and/or C_16:1 ω6c) and summed feature 4 (iso-C_17:1 I and/or anteiso-C_17:1 B). On the basis of the phylogenetic analysis, chemotaxonomic data, physiological characteristics, and DNA–DNA hybridization data, strain THG-T16^T represents a novel species of the genus Nibribacter, for which the name Nibribacter flagellatus sp. nov. is proposed. The type strain is THG-T16^T(=?KACC 19188^T?=?CCTCC AB 2016246^T).     

<Emphasis Type="Italic">Streptomyces caldifontis</Emphasis> sp. nov., isolated from a hot water spring of Tatta Pani,Kotli, Pakistan

A Gram-staining positive, non-motile, rod-shaped, catalase positive and oxidase negative bacterium, designated NCCP-1331^T, was isolated from a hot water spring soil collected from Tatta Pani, Kotli, Azad Jammu and Kashmir, Pakistan. The isolate grew at a temperature range of 18-40 °C (optimum 30 °C), pH 6.0–9.0 (optimum 7.0) and with 0–6 % NaCl (optimum 2 % NaCl (w/v)). The phylogenetic analysis based on 16S rRNA gene sequence revealed that strain NCCP-1331^T belonged to the genus Streptomyces and is closely related to Streptomyces brevispora BK160^T with 97.9 % nucleotide similarity, followed by Streptomyces drosdowiczii NRRL B-24297^T with 97.8 % nucleotide similarity. The DNA–DNA relatedness values of strain NCCP-1331^T with S. brevispora KACC 21093^T and S. drosdowiczii CBMAI 0498^T were 42.7 and 34.7 %, respectively. LL-DAP was detected as diagnostic amino acid along with alanine, glycine, leucine and glutamic acid. The isolate contained MK-9(H₈) as the predominant menaquinone. Major polar lipids detected in NCCP-1331^T were phosphatidylethanolamine, phosphatidylinositol and unidentified phospholipids. Major fatty acids were iso-C_{16: 0}, summed feature 8 (18:1 ω7c/18:1 ω6c), anteiso-C_15:0 and C_16:0. The genomic DNA G + C content was 69.8 mol %. On the basis of phylogenetic, phenotypic and chemotaxonomic analysis, it is concluded that strain NCCP-1331^T represents a novel species of the genus Streptomyces, for which the name Streptomyces caldifontis sp. nov. is proposed. The type strain is NCCP-1331^T (=KCTC 39537^T = CPCC 204147^T).     

Effect of low-energy microwaves on DNA stability in solution

DNA isolated from liver of healthy and tumor-bearing (sarcoma 45) rats was irradiated in water-salt solution with weak microwaves (64.5 GHz, 50 μW/cm²). The heat stability of DNA increased with irradiation time (a raise of 1.5°C in T _m for “tumor” DNA after 90 min, without changes in ΔT), which may be associated with dehydration of the surrounding Na⁺ ions.     

<Emphasis Type="Italic">Flavisolibacter carri</Emphasis> sp. nov., isolated from an automotive air-conditioning system

A Gram-stain negative, aerobic, non-motile, rod-shaped and yellow bacterium, designated TX0651^T, was isolated from an automotive air-conditioning system. Phylogenetically, the strain groups with the members of the genus Flavisolibacter and exhibits high 16S rRNA gene sequence similarities with Flavisolibacter ginsenosidimutans Gsoil 636^T (97.4%), Flavisolibacter ginsengiterrae Gsoil 492^T (96.3%) and Flavisolibacter ginsengisoli Gsoil 643^T (96.2%). DNA–DNA relatedness between TX0651^T and F. ginsenosidimutans KCTC 22818^T and F. ginsengiterrae KCTC 12656^T were determined to be less than 40%. The low levels of DNA–DNA relatedness identifies the strain TX0651^T as a novel species in the genus Flavisolibacter. The major cellular fatty acids were identified as iso-C_15:0, summed feature 3 (C_16:1 ω7c and/or C_16:1 ω6c), iso-C_15:1 G and iso-C_17:0 3-OH. The predominant respiratory quinone was identified as MK-7. The polar lipids were found to be comprised of phosphatidylethanolamine, unidentified amino-glycophospholipids, an unidentified aminophospholipid, an unidentified amino lipid and unidentified lipids. The DNA G+C content of the strain was determined to be 31.2 mol%. On the basis of the phenotypic, genotypic and chemotaxonomic characteristics, strain TX0651^T should be classified in a novel species in the genus Flavisolibacter, for which the name Flavisolibacter carri sp. nov. (=?KACC 19014^T?=?KCTC 52836^T?=?NBRC 111784^T) is proposed.     

A novel pigmented and heavy metal biosorptive bacterium, <Emphasis Type="Italic">Leucobacter epilobiisoli</Emphasis> sp. nov., isolated from rhizosphere soil of <Emphasis Type="Italic">Epilobium hirsutum</Emphasis> L.

A novel yellow pigmented, Gram-positive, aerobic and heavy metal biosorptive bacterium designated SYP-B2667^T was isolated from rhizosphere soil of Epilobium hirsutum L. in Tongren, Guizhou province, China. Based on 16S rRNA gene sequence analyses, it was shown that strain SYP-B2667^T represents a novel species in the genus Leucobacter, with Leucobacter chromiireducens subsp. solipictus JCM 15573^T as a close phylogenetic neighbour (sequence similarity of 98.2%). Chemotaxonomic characteristics also supported the affiliation to the genus Leucobacter. Strain SYP-B2667^T was determined to have a DNA G+C content of 66.6 mol%; 2,4-diaminobutyric acid in the cell wall peptidoglycan amino acids; MK-11 as predominant menaquinone; an abundance of anteiso-C_15:0 and anteiso-C_17:0 fatty acids; and polar lipids including diphosphatidylglycerol, phosphatidylglycerol, glycolipids and unidentified phospholipids. The DNA–DNA hybridization value between strain SYP-B2667^T and L. chromiireducens subsp. solipictus JCM 15573^T was 19.7?±?2.8%. Based on these phylogenetic and phenotypic results, it can be concluded that strain SYP-B2667^T represents a novel species, for which the name Leucobacter epilobiisoli sp. nov. is proposed. The type strain is SYP-B2667^T (=DSM 105145^T=CPCC 204976^T). This strain can tolerate and adsorb five heavy metals and so may have potential to facilitate heavy metal removal and bioremediation.     

<Emphasis Type="Italic">Siphonobacter intestinalis</Emphasis> sp. nov., a bacterium isolated from the feces of <Emphasis Type="Italic">Pseudorhynchus japonicus</Emphasis>

Strain 63MJ-2^T was isolated from the feces of broad-winged katydid (Pseudorhynchus japonicus) collected in Korea. The 16S rRNA gene sequence of this strain showed the highest sequence similarity with that of Siphonobacter aquaeclarae P2^T (96.1%) and had low similarities (below 86.3%) with those of other members of family ‘Flexibacteraceae’. The strain 63MJ-2^T is a strictly aerobic, Gram-stain-negative, non-motile, rod-shaped bacterium. The strain grew at 4–35°C (optimum, 25–30°C), pH of 5.0–9.0 (optimum, 6.0–7.0), and 0–2.0% (optimum, 1.0–2.0) (w/v) NaCl. The DNA G+C content of strain 63MJ-2^T was 43.5 mol%. The major fatty acids were C_16:1ω5c (42.5%), iso-C_17:0 3-OH (18.7%), and summed feature 3 (iso-C_15:0 2-OH and/or C_16:1ω7c, 18.0%). The major menaquinone was MK-7 and polar lipids were phosphatidylethanolamine, six unknown aminolipids, and five unknown lipids. Based on the evidence from our polyphasic taxonomic study, we conclude that strain 63MJ-2^T should be classified as a novel species of the genus Siphonobacter, and propose the name Siphonobacter intestinalis sp. nov. The type strain is 63MJ-2^T (=KACC 18663^T =NBRC 111883^T).     

<Emphasis Type="Italic">Actinoplanes deserti</Emphasis> sp. nov., isolated from a desert soil sample

A novel actinomycete, designated strain YIM CF22^T, was isolated from a desert soil sample collected from Turpan in Xinjiang Uyghur Autonomous Region, north-western China. The taxonomic position of the strain YIM CF22^T is described based on a polyphasic approach. Strain YIM CF22^T was found to form irregular sporangia on agar media. It contains meso-diaminopimelic acid in the cell wall peptidoglycan. The major menaquinone was identified as MK-9(H₄); the polar lipids were identified as diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, two unidentified phospholipids and two unidentified glycolipids. The whole cell sugars were found to be ribose, mannose, galactose, glucose and xylose. The major cellular fatty acids were found to be (>?5%) iso-C_16:0 (43.5%), anteiso-C_17:0 (10.2%), iso-C_15:0 (7.1%), C_17:1 ω8c (6.3%) and iso H-C_16:1 (5.9%). The G+C content was determined to be 70.8%. 16S rRNA gene sequence analysis of strain YIM CF22^T showed high similarity (97.0%) to Actinoplanes rishiriensis NBRC 108556^T. The strain also showed high 16S rRNA gene sequence similarities to Verrucosispora sediminis CGMCC 4.3550^T (96.9%) and Micromonospora tulbaghiae DSM 45142^T (96.8%). Phylogenetic analysis based on the 16S rRNA gene sequence indicated that strain YIM CF22^T clusters with A. rishiriensis NBRC 108556^T, Actinoplanes globisporus JCM 3186^T and Actinoplanes rhizophilus NEAU-A-2^T. Based on the differential phenotypic characteristics and the results of DNA–DNA relatedness and phylogenetic analysis, it is proposed that strain YIM CF22^T represents a novel species of the genus Actinoplanes, for which the name Actinoplanes deserti sp. nov. is proposed. The type strain is YIM CF22^T (=?KCTC 39543^T?=?CCTCC AB2018113^T).