V. Venkata Ramana

Rhizobium subbaraonis sp. nov., an endolithic bacterium isolated from beach sand.

Two strains (JC85(T) and JC108) of Gram-stain-negative, motile bacteria were isolated from endolithic beach sand samples on an oligotrophic medium. Based on the 16S rRNA gene sequence analysis, both strains were identified as belonging to the genus Rhizobium. Strain JC108 had 16S rRNA gene sequence similarity of 100 % with Rhizobium pusense NRCPB10(T) and formed a cluster with this strain. Strain JC85(T) had 96.9 % 16S rRNA gene sequence similarity and was 18 % related (based on DNA-DNA hybridization) to Rhizobium borbori DN316(T). With other strains of the genus Rhizobium, the 16S rRNA gene sequence similarity was less than 96.3 %. Strain JC85(T) could tolerate up to 3 % salinity, fix N(2), was resistant to ampicillin (10 µg) and was positive for catalase and oxidase. The major fatty acid was C(18 : 1)ω7c (69 %) with minor amounts of C(19 : 0) cyclo ω8c (8.9 %), C(16 : 0) (6.9 %), C(12 : 0) (5.7 %) and C(19 : 1)ω7c/C(19 : 1)ω6c (2.2 %). Polar lipids of strain JC85(T) include two unidentified aminophospholipids (APL1,2), two unidentified phospholipids (PL1,2), phosphatidylcholine and four unidentified lipids (L1-4). Q-10 is the major quinone of strain JC85(T). Based on polyphasic taxonomic analysis, strain JC85(T) represents a novel species for which, the name Rhizobium subbaraonis JC85(T) is proposed. The type strain is JC85(T) ( = DSM 24765(T) = KCTC 23614(T)).

Descriptions of Rhodopseudomonas parapalustris sp. nov., Rhodopseudomonas harwoodiae sp. nov. and Rhodopseudomonas pseudopalustris sp. nov., and emended description of Rhodopseudomonas palustris

Four strains (JA310(T), JA531(T), JA447 and JA490) of red to reddish brown pigmented, rod-shaped, motile and budding phototrophic bacteria were isolated from soil and freshwater sediment samples from different geographical regions of India. All strains contained bacteriochlorophyll a and carotenoids of the spirilloxanthin series. The major cellular fatty acid of strains JA310(T) and JA531(T) was C(18:1)ω7c, the quinone was Q-10 and polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, an aminohopanoid and an unidentified aminolipid. Phylogenetic analysis based on 16S rRNA gene sequences showed that all strains clustered with species of the genus Rhodopseudomonas in the class Alphaproteobacteria. Strains JA531(T), JA447 and JA490 were genotypically (>80% related based on DNA-DNA hybridization) and phenotypically closely related to each other and the three strains were distinct from strain JA310(T) (33% related). Furthermore, all four strains had less than 48% relatedness (DNA-DNA hybridization) with type strains of members of the genus Rhodopseudomonas, i.e. Rhodopseudomonas palustris ATCC 17001(T), Rhodopseudomonas faecalis JCM 11668(T) and Rhodopseudomonas rhenobacensis DSM 12706(T). The genomic DNA G+C contents of strains JA310(T) and JA531(T) were 63.8 and 62.4 mol%, respectively. On the basis of phenotypic, chemotaxonomic and molecular genetic evidence, it is proposed that strains JA310(T) ( = NBRC 106083(T) = KCTC 5839(T)) and JA531(T) ( = NBRC 107575(T) = KCTC 5841(T)) be classified as the type strains of two novel species of the genus Rhodopseudomonas, Rhodopseudomonas parapalustris sp. nov. and Rhodopseudomonas harwoodiae sp. nov., respectively. In addition, we propose that strain DSM 123(T) ( = NBRC 100419(T)) represents a novel species, Rhodopseudomonas pseudopalustris sp. nov., since this strain differs genotypically and phenotypically from R. palustris ATCC 17001(T) and other members of the genus Rhodopseudomonas. An emended description of R. palustris is also provided.

Roseomonas aestuarii sp. nov., a bacteriochlorophyll-a containing alphaproteobacterium isolated from an estuarine habitat of India.

Two strains (JC17(T) and JC19a) of orange pigmented bacteria were isolated from an estuarine sample. Cells of both the strains were Gram-negative coccobacilli, non-motile, non-spore forming and strictly aerobic. Chemo-organoheterotrophy was the growth mode for both strains and was possible on a wide range of organic compounds. Strains were non-hemolytic and contained low levels of BChl-a and carotenoids. The fatty acids (>1.0%) comprised C(18:1)omega7c, C(16:1)omega7c/iso-C(15:0)2OH, C(16:0), C(16:0) 3-OH, C(18:1)2OH, C(16:1)omega5c, and C(19:0) cycloomega8c. The genomic DNA G+C content of strain JC17(T) was 66.2mol%. A phylogenetic tree based on 16S rRNA gene sequence analysis showed that strains JC17(T) and JC19a had the highest similarity to members of the genus Roseomonas and were closely related to Roseomonas cervicalis CIP104027(T) (96.4%) and Roseomonas ludipueritiae CIP107418(T) (96.3%) of the family Acetobacteraceae within the class Alphaproteobacteria. Strains JC17(T) and JC19a shared 100% 16S rRNA gene sequence similarity, were phenotypically (morphological, physiological, biochemical characters) identical and had closely related genomes (85% DDH). Based on polyphasic taxonomic data, strain JC17(T) is classified as a novel species of the genus Roseomonas for which the name Roseomonas aestuarii sp. nov. is proposed. The type strain is JC17(T) (=CCUG 57456(T) =KCTC 22692(T) =NBRC105654(T)).

Rhodobacter aestuarii sp. nov., a phototrophic alphaproteobacterium isolated from an estuarine environment

An ovoid to rod-shaped, phototrophic, purple non-sulfur bacterium was isolated from a brown-coloured microbial mat from the brackish water of Bhitarkanika mangrove forest, Dangmal, Orissa, India. Cells of strain JA296(T) were Gram-negative and motile, forming chains of four to eight cells. The colour of the cell suspension grown under anaerobic conditions in the light was yellowish green. Bacteriochlorophyll a and the carotenoids spheroidene and spheroidenone of the spirilloxanthin series were present as photosynthetic pigments. The bacterium was a facultative anaerobe and was able to grow photo-organoheterotrophically and chemo-organoheterotrophically. Thiamine was required as a growth factor. C(18 : 1)omega7c was the dominant fatty acid. Internal cytoplasmic membranes were of the vesicular type. Strain JA296(T) did not require NaCl for growth. Phylogenetic analysis on the basis of 16S rRNA gene sequences showed that strain JA296(T) was most closely related to Rhodobacter capsulatus ATCC 11166(T) (95.5 % sequence similarity) and clustered with species of the genus Rhodobacter of the family Rhodobacteraceae, class Alphaproteobacteria. On the basis of 16S rRNA gene sequence analysis and morphological and physiological characteristics, strain JA296(T) represents a novel species of the genus Rhodobacter, for which the name Rhodobacter aestuarii sp. nov. is proposed; the type strain is JA296(T) (=JCM 14887(T) =CCUG 55130(T)).

Rhodobacter maris sp. nov., a phototrophic alphaproteobacterium isolated from a marine habitat of India

During investigations into the diversity of anoxygenic phototrophic bacteria in marine habitats, an ovoid to rod-shaped purple non-sulfur bacterium, designated strain JA276(T), was isolated from enrichments under photoheterotrophic conditions from a marine sediment sampled from the seashore of Cochin, India. Strain JA276(T) is a Gram-negative, motile, chain-forming bacterium that shows optimum growth under photoheterotrophic conditions and is also able to grow chemoorganotrophically. Thiamine is required as a growth factor. Strain JA276(T) contains vesicular intracytoplasmic membranes, bacteriochlorophyll a and the carotenoids spheroidene and spheroidenone. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain JA276(T) belongs to the genus Rhodobacter and is closely related to the type strain of Rhodobacter capsulatus (96.2 % sequence similarity). On the basis of the results of 16S rRNA gene sequence analysis and morphological and physiological data, strain JA276(T) is significantly different from other species of the genus Rhodobacter and represents a novel species of the genus, for which the name Rhodobacter maris sp. nov. is proposed. The type strain is JA276(T) (=JCM 14794(T) =ATCC BAA-1549(T) =CCUG 55129(T)).

Rhodobacter megalophilus sp. nov., a phototroph from the Indian Himalayas possessing a wide temperature range for growth

Two strains of phototrophic, purple non-sulfur bacteria capable of growing at low temperatures (5 degrees C) were isolated from the Himalayas. The two strains showed positive phototaxis and grew over a relatively wide temperature range (5-40 degrees C). Phylogenetic analysis based on 16S rRNA gene sequences showed that strain JA194T clustered with members of the genus Rhodobacter. Strain JA194T showed highest 16S rRNA gene sequence similarity with Rhodobacter sphaeroides DSM 158T (99 %). However, DNA-DNA hybridization experiments between Rba. sphaeroides DSM 158T and strain JA194T revealed a level of relatedness of only 67 %. The DNA base composition of strain JA194T was 66.67 mol% G+C (by HPLC). Based on 16S rRNA gene sequence analysis, morphological, physiological, Fourier transform infrared fingerprinting and DNA-DNA hybridization studies, strain JA194T (=KCTC 5602T =JCM 14598T) is sufficiently different from other Rhodobacter species to merit its description as the type strain of a novel species, for which the name Rhodobacter megalophilus sp. nov. is proposed.

Rhodomicrobium udaipurense sp. nov., a psychrotolerant, phototrophic alphaproteobacterium isolated from a freshwater stream

Two strains (JA643(T) and JA755) of Gram-stain-negative, facultatively anaerobic phototrophic, bacteria capable of growth at low temperatures (10-15 °C) were isolated from freshwater streams from different geographical regions of India. Both strains contain bacteriochlorophyll a and carotenoids of the spirilloxanthin series. Phosphatidylethanolamine, phosphatidylcholine, phosphatidylglycerol, diphosphatidylglycerol, an unidentified phospholipid (PL), unidentified amino lipids (AL1-AL6, AL9) and an unidentified lipid (L1) were the polar lipids present in both strains. The major cellular fatty acid was C18 : 1ω7c (76-79 % of the total). Bacteriohopane derivatives (BHD1,2), unidentified hopanoids (UH1-5), diplopterol (DPL) and diploptene (DPE) were the major hopanoids of both strains. The DNA G+C content was 64.2-64.5 mol%. 16S rRNA gene sequence-based phylogenetic analysis showed that both strains are closely related to the genus Rhodomicrobium and clustered with Rhodomicrobium vannielii DSM 162(T) (99 % sequence similarity). However, both strains exhibited only 46.1 % DNA-DNA hybridization with R. vannielii DSM 162(T). Strains JA643(T) and JA755 shared >99 % 16S rRNA gene sequence similarity and were >85 % related on the basis of DNA-DNA hybridization; they are therefore considered to represent a novel species in the genus Rhodomicrobium, for which the name Rhodomicrobium udaipurense sp. nov. is proposed. The type strain is JA643(T) ( = KCTC 15219(T) = NBRC 109057(T)).

Rhodococcus enclensis sp. nov., a novel member of the genus Rhodococcus

A novel actinobacterial strain, designated, NIO-1009(T), was isolated from a marine sediment sample collected from Chorao Island, Goa, India. Phylogenetic analysis comparisons based on 16S rRNA gene sequences between strain NIO-1009(T) and other members of the genus Rhodococcus revealed that strain NIO-1009(T) had the closest sequence similarity to Rhodococcus kroppenstedtii DSM 44908(T) and Rhodococcus corynebacterioides DSM 20151(T) with 99.2 and 99.1%, respectively. Furthermore, DNA-DNA hybridization results showed that R. kroppenstedtii DSM 44908(T) and R. corynebacterioides DSM 20151(T) were 39.5 (3.0%) and 41.7 (2.0%) with strain NIO-1009(T), respectively, which were well below the 70% limit for any novel species proposal. Phylogenetically strain NIO-1009(T) forms a stable clade with and R. kroppenstedtii DSM 44908(T) and R. corynebacterioides DSM 20151(T) with 100% bootstrap values. Strain NIO-1009(T) contained meso-diaminopimelic acid as the diagnostic diamino acid and galactose and arabinose as the cell wall sugars. The major fatty acids were C(16 : 0), C(18 : 1)ω9c, C(16 : 1)(ω6c and/or ω7c) and 10-methyl C(18 : 0). The only menaquinone detected was MK-8(H2), while the major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol, phosphatidylinositol mannoside and one unknown phospholipid. The G+C content of the genomic DNA was 66.9 mol%. The phenotypic and genotypic data showed that strain NIO-1009(T) warrants recognition as a novel species of the genus Rhodococcus for which the name Rhodococcus enclensis sp. nov., is proposed; the type strain is NIO-1009(T) ( = NCIM 5452(T) = DSM 45688(T)).

Rufibacter immobilis sp. nov., isolated from a high-altitude saline lake.

Two pinkish-red, Gram-stain-negative, non-motile aerobic bacterial strains (MCC P1(T) and MCC P2), capable of growing at low temperatures (15 °C), were isolated from water of a saline lake located in the western Himalayas of India. The strains were capable of growth in the presence of 0-2.0% NaCl and at pH 6.5-9.0. Phylogenetic analysis based on 16S rRNA gene sequences revealed the closest similarity of 96.3% to the type strain of the only species of the genus Rufibacter , Rufibacter tibetensis CCTCC AB 208084(T). Strains MCC P1(T) and MCC P2 shared 99.0% 16S rRNA gene sequence similarity and 88.6% DNA-DNA relatedness. The major cellular fatty acids were iso-C15 : 0, C17 : 1ω6c, summed feature 3 (C16 : 1ω6c/C16 : 1ω7c) and summed feature 4 (anteiso-C17 : 1 B/iso-C17 : 1 I). Predominant polar lipids were phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol. The respiratory quinone was MK-7. The DNA G+C content of the strains was 52.6-52.8 mol%. Based on morphological, physiological, chemotaxonomical and molecular characteristics, strains MCC P1(T) and MCC P2 represent a novel species of the genus Rufibacter , for which the name Rufibacter immobilis sp. nov. is proposed. The type strain is MCC P1(T) ( =MCC 2268(T) =CCTCC AB 2013351(T)).

Nitrincola alkalisediminis sp. nov., an alkaliphilic bacterium isolated from an alkaline lake

Two Gram-stain-negative, aerobic, alkaliphilic bacteria (strains MEB087T and MEB142) were isolated from sediment and water samples, respectively, collected from the alkaline Lonar Lake in Maharashtra, India. Strains MEB087T and MEB142 shared 99.8 % 16S rRNA gene sequence similarity and were 85 % related on the basis of DNA-DNA hybridization. The 16S rRNA gene sequences of both strains showed close relationship with the genus Nitrincola, and their closest neighbour was Nitrincola lacisaponensis 4CAT with 97.7 % sequence similarity. MEB087T and MEB142 exhibited only 45 % and 54 % DNA-DNA relatedness, respectively, with Nitrincola lacisaponensis DSM 16316T. Both strains were asporogenous, short, non-motile rods capable of utilizing a limited range of organic acids as sole carbon and energy sources. They were oxidase- and catalase-positive, able to reduce nitrate and nitrite; but unable to degrade DNA, urea, gelatin, casein or starch. They grew optimally at pH 9.5 (tolerating up to pH 11) and could withstand up to 0.6 M NaCl. The predominant cellular fatty acids were summed feature 8 comprising C18 : 1ω7c/C18 : 1ω6c (47-49 %) followed by summed feature 3 comprising C16 : 1ω7c/C16 : 1ω6c (28-32 %). The predominant polar lipids were diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylcholine. The DNA G+C content was 49.3-49.7 mol%. On the basis of the phylogenetic analysis and chemotaxonomic characteristics, strains MEB087T and MEB142 represent a novel species in the genus Nitrincola, for which the name Nitrincola alkalisediminis sp. nov. is proposed. The type strain is MEB087T ( = KCTC 42948T = JCM 19317T) with MEB142 ( = KCTC 42949 = JCM 19318) as an additional strain.