E. V. V. Ramaprasad

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.

Georgenia satyanarayanai sp. nov., an alkaliphilic and thermotolerant amylase-producing actinobacterium isolated from a soda lake.

A Gram-stain-positive, oxidase-negative, starch-hydrolysing, actinobacterium (strain JC82(T)) was isolated from a soda lake in Lonar, India. Based on 16S rRNA gene sequence similarity studies, strain JC82(T) belonged to the genus Georgenia and was most closely related to Georgenia muralis 1A-C(T) (96.8 %) and other members of the genus Georgenia (<96.5 %). The DNA G+C content of strain JC82(T) was 73.4 mol%. The cell-wall amino acids were alanine, glutamic acid and lysine with peptidoglycan type A4α. Polar lipids included diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, phosphatidylinositol mannosides, an unidentified lipid (L1) and an unidentified glycolipid (GL3). The predominant isoprenoid quinone was menaquinone MK-8(H(4)). Anteiso-C(15 : 0) was the predominant fatty acid and significant proportions of iso-C(14 : 0), C(14 : 0), C(16 : 0), iso-C(15 : 0) and iso-C(16 : 0) were also detected. Strain JC82(T) produced thermostable alkaline amylase. The results of physiological and biochemical tests allowed a clear phenotypic differentiation of strain JC82(T) from all other members of the genus Georgenia. Based on these data, strain JC82(T) represents a novel species of the genus Georgenia, for which the name Georgenia satyanarayanai sp. nov. is proposed. The type strain is JC82(T) ( = KCTC 19802(T) = NBRC 107612(T)).

Roseomonas oryzae sp. nov., isolated from paddy rhizosphere soil.

A non-motile, coccus-shaped, pale-pink-pigmented bacterium, designated strain JC288T, was isolated from a paddy rhizosphere soil collected from Western Ghats, Kankumbi, Karnataka, India. Cells were found to be Gram-stain-negative, and catalase- and oxidase-positive; the major fatty acids were C16 : 0, C16 : 1ω7c/C16 : 1ω6c, C18 : 1ω7c/C18 : 1ω6c and C18 : 1 2-OH. The predominant respiratory quinone was Q-10 and the genomic DNA G+C content was 67.5 mol%. Strain JC288T contained diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, four unidentified aminolipids, three unidentified phospholipids, two unidentified lipids, an aminophospholipid and a glycolipid. Hydroxyspirilloxanthin was the major carotenoid of strain JC288T. 16S rRNA gene sequence comparisons indicated that strain JC288T represents a member of the genus Roseomonas within the family Acetobacteraceae of the phylum Proteobacteria. Strain JC288T shared the highest 16S rRNA gene sequence similarity with Roseomonas rhizosphaerae YW11T (97.3 %), Roseomonas aestuarii JC17T (97.1 %), Roseomonas cervicalis CIP 104027T (95.9 %) and other members of the genus Roseomonas ( < 95.5 %). The distinct genomic difference and morphological, physiological and chemotaxonomic differences from the previously described taxa support the classification of strain JC288T as a representative of a novel species of the genus Roseomonas, for which the name Roseomonas oryzae sp. nov. is proposed. The type strain is JC288T ( = KCTC 42542T = LMG 28711T).

Rhodovulum bhavnagarense sp. nov., a phototrophic alphaproteobacterium isolated from a pink pond.

An oval to rod-shaped, Gram-stain-negative, phototrophic bacterium, strain JA738(T), was isolated from a sediment sample collected from a pink pond. Strain JA738(T) was non-motile and had vesicular-type intracellular photosynthetic membranes. Bacteriochlorophyll a and carotenoids of the spheroidene series were present as the major photosynthetic pigments. Strain JA738(T) required thiamine and pantothenate for growth. The major cellular fatty acids were C(18 : 1)ω7c, C(18 : 1)ω5c, C(18 : 0) and C(18 : 1)ω7c11-methyl; minor amounts of C(10 : 0) 3-OH and C(16 : 0) were also present. The major quinone was Q-10 and major polar lipids were phosphatidylglycerol, phosphatidylethanolamine and two unidentified sulfolipids (SL1-2). Phylogenetic analysis on the basis of 16S rRNA gene sequences showed that strain JA738(T) clustered with species of the genus Rhodovulum in the class Alphaproteobacteria. Strain JA738(T) was most closely related to Rhodovulum adriaticum DSM 2781(T) (96.4 % 16S rRNA gene sequence similarity) and other members of the genus Rhodovulum (<96.1 %). On the basis of phenotypic and molecular genetic evidence, it is proposed that strain JA738(T) should be classified as a novel species of the genus Rhodovulum for which the name Rhodovulum bhavnagarense sp. nov. is proposed. The type strain is JA738(T) ( = DSM 24766(T) = KCTC 15110(T)).

Flectobacillus rhizosphaerae sp. nov., isolated from the rhizosphere soil of Oryza sativa (L.), and emended description of the genus Flectobacillus.

A Gram-stain-negative, aerobic, pale orange, rod-shaped, non-motile bacterial strain, designated JC289T was isolated from a rhizosphere soil sample of the plant Oryza sativa (L.). 16S rRNA gene sequence analysis clearly allocated strain JC289T to the Flectobacillus cluster, showing highest sequence similarities to Flectobacillus roseus GFA-11T (99.5 %), Flectobacillus major ATCC 29496T (96.7 %) and Flectobacillus lacus CL-GP79T (94.6 %). Major (>5 %) fatty acids of strain JC289T were C16 : 1ω5c, iso-C15 : 0, C15 : 0 2-OH, iso-C15 : 0 3-OH and C16 : 1ω7c/C16 : 1ω6c, with minor amounts (>1- < 5 %) of C14 : 0, C16 : 0, anteiso-C15 : 0, C16 : 0 3-OH, iso-C17 : 0 3-OH, anteiso-C17 : 1B and/or iso-C17 : 1I. Menaquinone-7 was the major quinone of strain JC289T, and the polar lipid profile contained phosphatidylethanolamine, five unidentified aminophospholipids, two unidentified phospholipids, two unidentified aminolipids, two unidentified glycolipids and five unidentified lipids. DNA-DNA relatedness values of strain JC289T with Flectobacillus roseus LMG 24501T and Flectobacillus major LMG 13163T were 24.6 % (31.4 %, reciprocal analysis) and 16.2 % (22.4 %), respectively. On the basis of 16S rRNA gene sequence analysis, physiological and biochemical test results, and chemotaxonomic analysis, strain JC289T can be differentiated from its closest relatives in the genus Flectobacillus. Based on the data presented, it is concluded that strain JC289T represents a novel species of the genus Flectobacillus, for which the name Flectobacillus rhizosphaerae sp. nov. is proposed. The type strain is JC289T ( = KCTC 42575T = LMG 28712T).

Chryseomicrobium amylolyticum sp. nov., isolated from a semi-arid tropical soil, and emended descriptions of the genus Chryseomicrobium and Chryseomicrobium imtechense

A rod-shaped, non-motile, Gram-stain-positive, catalase-positive, starch-hydrolysing strain, JC16(T), was isolated from a semi-arid tropical soil from India. Phylogenetic analysis based on 16S rRNA gene sequence showed that strain JC16(T) clustered with the type species of the genus Chryseomicrobium, Chryseomicrobium imtechense MW 10(T), a member of the family Planococcaceae within the phylum Firmicutes with 99.3 % sequence similarity. Major (>10 %) fatty acids of strain JC16(T) were iso-C15 : 0 and iso-C16 : 0. Minor (<10 and >1 %) amounts of C16 : 0, iso-C14 : 0, anteiso-C15 : 0, iso-C17 : 0, anteiso-C17 : 0, iso-C17 : 1ω10c and C16 : 1ω11c are present in strain JC16(T). Polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phospholipids (PL2-4), aminolipids (AL1, 2) and an unknown lipid. Cell wall peptidoglycan was of the type l-Orn-D-Glu. The quinone system was composed of MK-7, MK-8 and MK-6. Genomic DNA G+C content of strain JC16(T) was 57.6 mol%. Distinct physiological, chemotaxonomic and genotypic differences (37 % reassociation based on DNA-DNA hybridization) from Chryseomicrobium imtechense MW 10(T) support the classification of strain JC16(T) as a representative of a novel species in the genus Chryseomicobium, for which the name Chryseomicrobium amylolyticum sp. nov. (type strain JC16(T) = DSM 23442(T) = NBRC 105215(T)) is proposed.

Marichromatium litoris sp. nov. and Marichromatium chrysaorae sp. nov. isolated from beach sand and from a jelly fish (Chrysaora colorata)

Three strains (JA349(T), JA553(T), JA439) of phototrophic sulphur bacteria were isolated from marine habitats of India. 16S rRNA gene sequence of the three strains clustered phylogenetically with members of the genus Marichromatium of the family Chromatiaceae belonging to the class Gammaproteobacteria. All the strains shared highest sequence similarity with the type strains of Marichromatium spp. (96-99% sequence similarity) and the new strains were characterized based on polyphasic taxonomy. Strains JA349(T) and JA553(T) can be distinguished from closest relative species of the genus Marichromatium with respect to distinct differences in cellular polar lipids, fatty acids and carbon/nitrogen sources utilization. Both strains were distinctly related (<50% based on DNA-DNA hybridization) with the type strains of the genus Marichromatium. Multilocus Sequence Analysis (MLSA) of the concatenated five protein coding genes (fusA, pufM, dnaK, recA, soxB) along with internal transcribed spacer (ITS; 16S-23S rRNA) had sequence similarity of less than 92% with the type strains of Marichromatium spp. Distinct phenotypic, chemotaxonomic and molecular differences allow the separation of strains JA349(T) and JA553(T) into new species of the genus Marichromatium for which, we propose the names Marichromatium litoris sp. nov. and Marichromatium chrysaorae sp. nov., respectively.

Rhodospirillum oryzae sp. nov., a phototrophic bacterium isolated from rhizosphere soil of paddy.

A reddish-brown bacterium, designated strain JA318(T), was purified from a photoheterotrophic enrichment culture obtained from the rhizosphere soil of paddy. Cells of strain JA318(T) are spiral shaped, Gram-stain-negative and motile by means of amphitrichous flagella. Strain JA318(T) has no NaCl requirement for growth but can tolerate up to 1.5 % (w/v) NaCl. Internal photosynthetic membranes are present as lamellar stacks. Photoorganoheterotrophy is the only growth mode observed. Strain JA318(T) contains bacteriochlorophyll a, lycopene and rhodopin as major carotenoids. Thiamine, niacin and para-aminobenzoic acid (PABA) are required as growth factors. Major fatty acids are C18 : 1ω7c and C16 : 0. Ubiquinone-8 and rhodoquinone-8 are the observed quinones. Diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and an unidentified aminolipid are the major polar lipids in strain JA318(T). Phylogenetic analysis based on 16S rRNA gene sequences showed that the strain JA318(T) clustered with species of the genus Rhodospirillum which belongs to the class Alphaproteobacteria. The highest sequence similarity of strain JA318(T) was found with Rhodospirillum sulfurexigens JA143(T) (99.9 %). The DNA-DNA reassociation values of strain JA318(T) with Rsp. sulfurexigens JA143(T) and Rhodospirillum photometricum DSM 122(T) were 52 ± 2 % and 45 ± 1 %, respectively. The genomic DNA G+C content of strain JA318(T) was 60.2 mol%. Based on the morphological, physiological, chemotaxonomical and molecular evidence, strain JA318(T) is significantly different from the type strains of species of the genus Rhodospirillum, of the family Rhodospirillaceae, and it is proposed that the strain be classified as a representative of a novel species for which the name Rhodospirillum oryzae sp. nov. is proposed. The type strain is JA318(T) (= KCTC 5960(T) = NBRC 107573(T)).

Emended description of the genus Rhodothalassium Imhoff et al., 1998 and proposal of Rhodothalassiaceae fam. nov. and Rhodothalassiales ord. nov

Five strains (JA325, JA389, JA473, JA563 and JA582) of Gram stain-negative, vibrioid to spiral shaped, phototrophic purple bacteria were isolated from solar salterns of India. All strains contained bacteriochlorophyll-a and carotenoids of the spirilloxanthin series as photosynthetic pigments. C(18:1)ω7c, C(18:1)ω7c 11-methyl and C(16:0) were the major fatty acids of all strains. Diphosphatidylglycerol (DPG), phosphatidylglycerol (PG), ornithine lipid (OL), an unidentified phospholipid (PL), and an unidentified aminolipid (AL) were the major polar lipids of all the strains. According to 16S rRNA gene sequences, all strains clustered phylogenetically with the only species of the genus Rhodothalassium (99.8-99.3% sequence similarity) but only strains JA325 and JA563 were distinctly related (60+1.5% DNA-DNA hybridization [DDH]) to the type strain Rhodothalassium salexigens DSM 2132(T). However, the genotypic data of strains JA325 and JA563 was not supported because of a large number of phenotypic differences compared to the type strain, therefore, it is proposed that all five newly isolated strains were R. salexigens-like strains. In addition, phylogenetically, the Rhodothalassium clade represented a distinct lineage and formed a deep branch with less than 90% 16S rRNA gene sequence similarity to other orders of the Alphaproteobacteria, and characteristic phenotypic properties also distinguished these bacteria from other purple non-sulfur bacteria. Therefore, the novel family Rhodothalassiaceae fam. nov. and the novel order Rhodothalassiales ord. nov. are proposed for the distinct phyletic line represented by the genus Rhodothalassium.