Subrata K. Das

Rhizobium pusense sp. nov., isolated from the rhizosphere of chickpea (Cicer arietinum L.).

A novel bacterial strain, designated NRCPB10(T), was isolated from rhizosphere soil of chickpea (Cicer arietinum L.) in Pusa, New Delhi, India. The 16S rRNA gene sequence of strain NRCPB10(T) showed highest similarity (98.9 %) to that of Rhizobium radiobacter NCPPB 2437(T), followed by Rhizobium larrymoorei AF3-10(T) (97.7 %) and Rhizobium rubi IFO 13261(T) (97.4 %). Phylogenetic analysis of strain NRCPB10(T) based on the housekeeping genes recA and atpD confirmed its position as distinct from recognized Rhizobium species. Levels of DNA-DNA relatedness between strain NRCPB10(T) and R. radiobacter ICMP 5785(T), R. larrymoorei LMG 21410(T) and R. rubi ICMP 6428(T) were 51.0, 32.6 and 27.3 %, respectively. Cellular fatty acids of strain NRCPB10(T) were C(18 : 1)ω7c (58.9 %), C(16 : 0) (15.5 %), C(19 : 0) cyclo ω8c (11.5 %), iso-C(16 : 1) (5.8 %), C(16 : 0) 3-OH (4.5 %), C(16 : 1)ω7c (2.1 %) and C(18 : 0) (1.3 %). The G+C content of the genomic DNA of strain NRCPB10(T) was 59.0 mol%. Strain NRCPB10(T) did not nodulate chickpea plants or induce tumours in tobacco plants. Phenotypic and physiological properties along with SDS-PAGE of whole-cell soluble proteins differentiated strain NRCPB10(T) from its closest phylogenetic neighbours. On the basis of data from the present polyphasic taxonomic study, strain NRCPB10(T) is considered to represent a novel species of the genus Rhizobium, for which the name Rhizobium pusense sp. nov. is proposed. The type strain is NRCPB10(T) ( = LMG 25623(T) = JCM 16209(T) = NCIMB 14639(T)).

Chelatococcus sambhunathii sp. nov., a moderately thermophilic alphaproteobacterium isolated from hot spring sediment

A moderately thermophilic bacterial strain (HT4(T)) isolated from a hot spring sediment was characterized phenotypically and phylogenetically. Cells were Gram-negative, aerobic, non-sporulating, rod-shaped and motile by means of a single polar flagellum. Both oxidase and catalase activities were positive. Heterotrophic growth was observed at pH 6.0-8.5 and at 20-50 degrees C; optimum growth occurred at pH 7.5-8.0 and 37-42 degrees C. The major cellular fatty acids were C( 14 : 0) 3-OH, C(18 : 0) 3-OH, C( 18 : 1) 2-OH, C(18 : 1)omega 7c and C(19 : 0) cyclo omega8 c. The DNA G+C content of strain HT4(T) was 67.8 mol%. 16S rRNA gene sequence analysis indicated that strain HT4(T) clustered within the radiation of the genus Chelatococcus and showed 99.0 % similarity with Chelatococcus daeguensis CCUG 54519(T) and 96 % similarity with Chelatococcus asaccharovorans DSM 6462(T). However, levels of DNA-DNA relatedness between strain HT4(T) and Chelatococcus daeguensis CCUG 54519(T) and Chelatococcus asaccharovorans DSM 6462(T) were 52 and 20%, respectively. On the basis of the phenotypic, physiological and chemotaxonomic data, 16S rRNA gene sequence analysis and DNA-DNA hybridization results, strain HT4(T) is considered to represent a novel species of the genus Chelatococcus, for which the name Chelatococcus sambhunathii sp. nov. is proposed. The type strain is HT4(T) (=DSM 18167(T)=JCM 14988(T)).

Marinomonas fungiae sp. nov., isolated from the coral Fungia echinata from the Andaman Sea.

A novel aerobic marine bacterium, strain AN44(T), was isolated from the coral Fungia echinata sampled from the Andaman Sea, India. Cells were Gram-negative, motile and rod-shaped. Oxidase and catalase tests were positive. Heterotrophic growth was observed at pH 5.5-10 and at 16-42 °C, with optimum growth at pH 7-8 and 28 °C. Strain AN44(T) grew in the presence of 0.5-11% (w/v) NaCl; the optimal NaCl concentration for growth was 3-5%. The DNA G+C content was 47.8 mol%. Predominant cellular fatty acids of strain AN44(T) were C(18 : 1)ω7c, C(16 : 1)ω7c/C(16 : 1)ω6c, C(16 : 0), C(10 : 0) 3-OH, C(12 : 0), C(10 : 0), C(14 : 0) and C(18 : 0). The sole isoprenoid ubiquinone was Q-8. The polar lipids were an unidentified phospholipid, an unidentified aminophospholipid and two unidentified glycolipids. 16S rRNA gene sequence comparisons revealed that strain AN44(T) clustered within the radiation of the genus Marinomonas and showed similarity of 97.9% with Marinomonas ostreistagni UST010306-043(T), 97.8% with Marinomonas aquimarina 11SM4(T), 97.1% with Marinomonas brasilensis R-40503(T) and 97.0% with Marinomonas communis 8(T). However, DNA-DNA relatedness between strain AN44(T) and closely related type strains was well below 70%. On the basis of the data from the present polyphasic taxonomic study, strain AN44(T) is considered to represent a novel species of the genus Marinomonas, for which the name Marinomonas fungiae sp. nov. is proposed. The type strain is AN44(T) (u200a=u200aJCM 18476(T)u200a=u200aLMG 27065(T)).

Taxonomic study of the genus Tepidiphilus: transfer of Petrobacter succinatimandens to the genus Tepidiphilus as Tepidiphilus succinatimandens comb. nov., emended description of the genus Tepidiphilus and description of Tepidiphilus thermophilus sp. nov., isolated from a terrestrial hot spring.

Comparative phenotypic, chemotaxonomic and genetic analysis revealed significant similarities among strains of the genera Tepidiphilus and Petrobacter. Analysis of 16S rRNA gene sequences and DNA-DNA relatedness of the type strains Tepidiphilus margaritifer N2-214(T) and Petrobacter succinatimandens 4BON(T) showed sequence similarity of 98.9u200a% and less than 40u200a% relatedness, indicating that these strains represent different species of same genus. Both strains had phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine and diphosphatidylglycerol as major polar lipids. Their fatty acid profiles were almost identical, with the predominant fatty acids C16u200a:u200a0, C17u200a:u200a0 cyclo and C19u200a:u200a0 cyclo ω8c. In view of this, we propose to transfer the member of the genus Petrobacter to the genus Tepidiphilus as Tepidiphilus succinatimandens comb. nov. and to emend the description of the genus Tepidiphilus. Further, a novel bacterium, strain JHK30(T), was isolated from a terrestrial hot spring located at Jharkhand, India, and was identified following a polyphasic approach. Cells were non-sporulating, aerobic, Gram-stain-negative rods and motile by a single polar flagellum. Optimum temperature for growth was 50-55 °C at pH 6.5-7.0. 16S rRNA gene sequence analysis revealed 99.71u200a% similarity with P. succinatimandens 4BON(T) (u200a=u200aDSM 15512(T)) and 98.71u200a% with T. margaritifer N2-214(T) (u200a=u200aDSM 15129(T)). However, DNA-DNA relatedness of strain JHK30(T) with these two type strains was well below 70u200a%. The DNA G+C base composition was 66.1 mol%. Strain JHK30(T) represents a novel species of the genus Tepidiphilus for which the name Tepidiphilus thermophilus sp. nov. is proposed. The type strain is JHK30(T) (u200a=u200aJCM 19170(T)u200a=u200aLMG 27587(T)= DSM 27220(T)).

Gulbenkiania indica sp. nov., isolated from a sulfur spring.

A novel bacterium, designated strain HT27(T), was isolated from a sulfur spring sample collected from Athamallik, Orissa, India, and was characterized by using a polyphasic approach. Cells were Gram-negative, strictly aerobic, rod-shaped and motile by means of a single polar flagellum. Strain HT27(T) was oxidase- and catalase-positive. Growth was observed at pH 5.0-11.0 and at 15-45 degrees C; the highest growth yield was observed at pH 7.5-8.0 and 30-37 degrees C. The G+C content of the genomic DNA of strain HT27(T) was 63 mol%. The major cellular fatty acids were C(16 : 1)omega7c (44.24 %), C(16 : 0) (27.65 %), C(18 : 1)omega7c (13.98 %), C(12 : 0) (2.60 %) and C(12 : 0) 3-OH (2.22 %). 16S rRNA gene sequence analysis indicated that strain HT27(T) clustered with the genus Gulbenkiania and showed 99.0 % similarity to Gulbenkiania mobilis E4FC31(T). However, the level of DNA-DNA relatedness between strain HT27(T) and G. mobilis E4FC31(T) was 30 %. On the basis of phenotypic and chemotaxonomic characteristics, 16S rRNA gene sequence analysis and DNA-DNA hybridization data, strain HT27(T) is considered to represent a novel species of the genus Gulbenkiania, for which the name Gulbenkiania indica sp. nov. is proposed. The type strain is HT27(T) (=DSM 17901(T) =JCM 15969(T)).

Comparative analysis of 16S rRNA signature sequences of the genus Idiomarina and Idiomarina woesei sp. nov., a novel marine bacterium isolated from the Andaman Sea

A Gram-negative, short rod, aerobic bacterium, designated W11(T), was isolated from seawater. Heterotrophic growth was observed at 10-45xa0°C and pH 6-10. Optimal growth was observed at 30-37xa0°C and pH 7-9. It can grow in the presence of 0.5-12% NaCl (w/v), and the optimal NaCl required for growth was 5-6%. 16S rRNA gene sequence similarity revealed that strain W11(T) clustered within the radiation of the genus Idiomarina and showed 99.24% similarity with Idiomarina donghaiensis JCM 15533(T), 97.64% with Idiomarina marina JCM 15083(T), 97.37% with Idiomarina tainanensis JCM 15084(T) and 97.16% with Idiomarina maritima JCM 15534(T). DNA-DNA similarities between strains W11(T) with other closely related strains were below 70%. Polar lipids included a phosphatidylgylycerol, a diphosphatidylglycerol, a phosphatidylethanolamine, an unidentified phosopholipid, two unidentified aminolipids and two unidentified lipids. DNA Gxa0+xa0C content was 41.2xa0±xa00.1xa0mol%. Major fatty acids were iso-C15:0, iso-C17:0, iso-C17:1ω9c, C16:0, iso-C11:0 3OH and C16:1ω7c/C16:1ω7c. The isoprenoid ubiquinone was Q8. On the basis of the present polyphasic taxonomic study, strain W11(T) is considered to represent a novel species of the genus Idiomarina, for which the name Idiomarina woesei sp. nov. is proposed. The type strain is W11(T) (=xa0DSM 27808(T)xa0=xa0JCM 19499(T)xa0=xa0LMG 27903(T)).

Microbacterium oryzae sp. nov., an actinobacterium isolated from rice field soil

A novel aerobic soil actinobacterium (strain MB10(T)) belonging to the genus Microbacterium was isolated from rice field soil samples collected from Jagatpur, Orissa, India. Cells were Gram-stain positive, short rod-shaped and motile. The strain was oxidase-negative and catalase-positive. Heterotrophic growth was observed at pH 5.0-11.0 and at 16-37 °C; optimum growth was observed at 28 °C and pH 7.0-9.0. The DNA G+C content was 71.6 mol%. Predominant cellular fatty acids of strain MB10(T) were iso-C14 : 0, anteiso-C15 : 0, C16 : 0, iso-C16 : 0 and anteiso-C17 : 0. Cell wall sugars were galactose, glucose and rhamnose. The major isoprenoid quinones were MK-9 (10 %), MK-10 (43 %) and MK-11 (36 %). The peptidoglycan represents the peptidoglycan type B2β. The polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phospholipid and unknown glycolipids. 16S rRNA gene sequence identity revealed the strain MB10(T) clustered within the radiation of the genus Microbacterium and showed 99.2 % similarity with Microbacterium barkeri DSM 20145(T). However, DNA-DNA similarity study was 37.0 % with Microbacterium barkeri DSM 20145(T), the nearest phylogenetic relative. On the basis of phenotypic and chemotaxonomic properties, 16S rRNA gene sequence analysis and DNA-DNA reassociation studies, it is proposed that strain MB10(T) represents a novel species of the genus Microbacterium, for which the name Microbacterium oryzae sp. nov. is proposed; the type strain is MB10(T) ( = JCM 16837(T) = DSM 23396(T)).

Vibrio panuliri sp. nov., a marine bacterium isolated from spiny lobster, Panulirus penicillatus and transfer of Vibrio ponticus from Scophthalmi clade to the newly proposed Ponticus clade

A novel marine bacterium, strain LBS2(T) was isolated from eggs carried on pleopods of the spiny lobster collected from Andaman Sea. Heterotrophic growth occurred at 1-7% NaCl. 16S rRNA gene sequence similarity revealed the strain LBS2(T) belonged to the genus Vibrio and showed above 97% similarity with eight type strains of the genus Vibrio. Multilocus analysis based on ftsZ, gapA, gyrB, mreB, pyrH recA, rpoA, and topA revealed LBS2(T) formed a separate cluster with Vibrio ponticus DSM 16217(T) with 89.8% multilocus gene sequence similarity. However, strain LBS2(T) is distantly related with other members of the Scophthalmi clade in terms of 16S rRNA signatures, phenotypic variations and multilocus gene sequence similarity, for which we propose LBS2(T) belongs to a new clade i.e. Ponticus clade with V. ponticus DSM 16217(T) as the representative type strain of the clade. DNA-DNA homologies between strain LBS2(T) and closely related strains were well below 70%. DNA Gxa0+xa0C content was 45.3xa0mol%. On the basis of our polyphasic study, strain LBS2(T) represents a novel species of the genus Vibrio, for which the name Vibrio panuliri sp. nov. is proposed. The type strain is LBS2(T) (= JCM 19500(T)xa0=xa0DSM 27724(T)xa0=xa0LMG 27902(T)).

Psychrobacter pocilloporae sp. nov., isolated from coral Pocillopora eydouxi from Andaman Sea, India.

Two closely related aerobic, Gram-stain-negative, rod-shaped bacteria (S6-60T and S6-67) were isolated from the mucus of the coral, Pocilloporaeydouxi, from the Andaman Sea, India. Heterotrophic growth on marine agar was observed at 4-37u2009°C and at pH 6.5-10.0; optimum growth occurred at 25-30u2009°C and at pH 7-9. 16S rRNA gene sequence analysis confirmed that the isolates belong to the genus Psychrobacter; the two isolates shared more than 99.5u2009% pairwise sequence similarity. Strain S6-60T showed a maximum 16S rRNA similarity of 98.92u2009% with Psychrobacter pacificensis DSM 23406T. DNA-DNA homology between the two isolates, S6-60T and S6-67, was above 90u2009%, whereas strain S6-60T showed less than 70u2009% homology with closely related type species. The DNA G+C content was 47.7 mol%. It contained phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine and phospholipid as the major polar lipids. C10u2009:u20090, C12u2009:u20090 3OH, C16u2009:u20090, C18u2009:u20091ω9c, C17u2009:u20091ω8c and C16u2009:u20091ω7c were found to be the predominant fatty acids. Based on a polyphasic analysis, the isolates (S6-60T and S6-67) represent a novel species of the genus Psychrobacter for which the name Psychrobacter pocilloporae sp. nov. is proposed with S6-60T(=JCM 31058T=LMG 29157T) as the type strain.

Sulfitobacter pontiacus subsp. fungiae subsp. nov., Isolated from Coral Fungia seychellensis from Andaman Sea, and Description of Sulfitobacter pontiacus subsp. pontiacus subsp. nov.

Two closely related aerobic, Gram reaction-negative rod-shaped bacteria (S7-75T and S7-80) were isolated from mucus of coral Fungia seychellensis from Andaman Sea, India. Heterotrophic growth on marine agar was observed at 4–35u2009°C and pH 6.5–10.5; optimum growth occurred at 25–30u2009°C and pH 7–8. 16xa0S rRNA sequence analysis confirmed the strains belonged to the genus Sulfitobacter and the two isolates shared more than 99.28% pairwise sequence similarity. DNA–DNA similarity between two isolates S7-75T and S7-80 was above 96%. Strain S7-75T showed maximum 16S rRNA similarity of 99.64% with Sulfitobacter pontiacus LMG 19752T. However, DNA–DNA relatedness between strain S7-75T and S. pontiacus LMG 19752T confirmed the placement of strain S7-75T as subspecies under the species S. pontiacus. Further, pulsed-field gel electrophoresis (PFGE), REP-PCR, ERIC-PCR fingerprint patterns and lipid profiles also differentiated strain S7-75T from the reference strain of S. pontiacus LMG 19752T. The DNA G+C content was 59.8xa0mol%. Q10 was the major respiratory quinone. Based on polyphasic analysis, the isolate S7-75T represents a subspecies of S. pontiacus for which the name S. pontiacus subsp. fungiae subsp. nov. is proposed with S7-75T (=JCM 31094Tu2009=u2009LMG 29158T) as type strain.