<em>Marichromatium indicum</em> sp. nov., a new purple sulfur Gammaproteobacterium from mangroves of Goa, India

A reddish-brown bacterium was isolated from photoheterotrophic enrichments of mangrove soil from the western coast of India, in a medium that contained 10 % (w/v) NaCl. Phylogenetic analysis on the basis of 16S rRNA gene sequences showed that strain JA100T clusters with species of the genus Marichromatium of the class ‘Gammaproteobacteria’. Cells of strain JA100T are Gram-negative, motile rods with monopolar single flagella; they require NaCl, the optimum concentration being 1–4 %, and tolerate concentrations up to 13 %. The strain has vesicular internal membrane structures, bacteriochlorophyll a and, most probably, carotenoids of the spirilloxanthin series. No growth factors are required. A reduced sulfur source is required for growth, and, during growth on reduced sulfur sources as electron donors, sulfur is intermediately deposited as a single large granule within the cell. Strain JA100T could not grow at the expense of other tricarboxylic acid cycle intermediates, except malate. On the basis of 16S rRNA gene sequence analysis and its morphological and physiological characteristics, strain JA100T is sufficiently different from other Marichromatium species to justify its designation as a novel species, for which the name Marichromatium indicum sp. nov. is proposed. The type strain is JA100T (=DSM 15907T=ATCC BAA-741T=JCM 12653T)

Rhodobium gokarnense sp. nov., a novel phototrophic alphaproteobacterium from a saltern

A pink-pigmented, phototrophic, purple nonsulfur bacterium, strain JA173T, was isolated in pure culture from a saltern in Gokarna, India, in a medium containing 2 % (w/v) NaCl. Strain JA173T was a non-motile Gram-negative rod that multiplied by budding. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain JA173T clusters with the class Alphaproteobacteria; highest sequence similarity (98 %) was to the type strain of Rhodobium orientis and 94 % similarity was observed to the 16S rRNA gene sequence of the type strain of Rhodobium marinum. However, DNA–DNA hybridization with R. orientis DSM 11290T revealed a relatedness value of only 35.1 % with strain JA173T. Strain JA173T contained lamellar internal membranes, bacteriochlorophyll a and carotenoids of the spirilloxanthin series. Strain JA173T had an obligate requirement for NaCl (optimum growth at 2–6 %, w/v) and grew photoheterotrophically with a number of organic compounds as carbon source or electron donor. Photoautotrophic, chemoautotrophic and fermentative growth could not be demonstrated. Yeast extract was required for growth. Based on 16S rRNA gene sequence analysis, DNA–DNA hybridization data and morphological and physiological characteristics, strain JA173T is sufficiently different from other species of the genus Rhodobium to be recognized as a representative of a novel species, Rhodobium gokarnense sp. nov. The type strain is JA173T (=ATCC BAA-1215T=DSM 17935T=JCM 13532T)

Thiophaeococcus mangrovi gen. nov., sp. nov., a photosynthetic marine gammaproteobacterium isolated from Bhitarkanika mangrove forest, India

A coccoid, phototrophic purple sulfur bacterium was isolated in pure culture from a mud sample collected from brackish water in the Bhitarkanika mangrove forest of Orissa, India, in a medium containing 2 % NaCl (w/v). This bacterium, strain JA304T, was Gram-negative and had a requirement for NaCl. Intracellular photosynthetic membranes were of the vesicular type. The colour of the phototrophically grown culture was saddle-brown. Bacteriochlorophyll a and the carotenoid lycopene were present as photosynthetic pigments. Strain JA304T was able to grow photolithoautotrophically and could photoassimilate a number of organic substrates. Yeast extract was required for growth of strain JA304T. The DNA G+C content was 68.1–68.9 mol%. 16S rRNA gene sequence comparisons indicate that the isolate represents a member of the Chromatiaceae within the class Gammaproteobacteria. According to sequence comparison data, strain JA304T is positioned distinctly outside the group formed by the four genera Thiocystis, Chromatium, Allochromatium and Thermochromatium, with only 86.7–91.0 % sequence similarity. Distinct morphological, physiological and genotypic differences from these previously described taxa support the classification of this isolate as a representative of a novel species in a new genus, for which the name Thiophaeococcus mangrovi gen. nov., sp. nov. is proposed. The type strain of Thiophaeococcus mangrovi is JA304T (=JCM 14889T =DSM 19863T). PABA, para-aminobenzoic acid The GenBank/EMBL/DDBJ accession number for the 16S rRNA gene sequence of strain JA304T is AM748925. A phase-contrast micrograph and whole-cell absorption spectrum and an acetone spectrum of extracted pigments of strain JA304T are available as supplementary material with the online version of this paper

Marichromatium bheemlicum sp. nov., a non-diazotrophic photosynthetic gammaproteobacterium from a marine aquaculture pond

A rod-shaped, phototrophic, purple sulfur bacterium, strain JA124(T), was isolated in pure culture from a marine aquaculture pond, located near Bhimunipatnam, in a medium that contained 3 % NaCl (w/v). Strain JA124(T) is a Gram-negative, motile rod with a single polar flagellum. Strain JA124(T) has a requirement for NaCl, with optimum growth at 1.5-8.5 %, and tolerates up to 11 % NaCl. Intracellular photosynthetic membranes are of the vesicular type. Bacteriochlorophyll a and probably carotenoids of the spirilloxanthin series are present as photosynthetic pigments. Strain JA124(T) was able to utilize sulfide, sulfate, thiosulfate, sulfite, thioglycollate and cysteine as sulfur sources. Strain JA124(T) was able to grow photolithoautotrophically, photolithoheterotrophically and photo-organoheterotrophically. Chemotrophic and fermentative growth could not be demonstrated. Strain JA124(T) lacks diazotrophic growth and acetylene reduction activity. Pyridoxal phosphate is required for growth. During growth on reduced sulfur sources as electron donors, sulfur is deposited intermediately as a number of small granules within the cell. Phylogenetic analysis on the basis of 16S rRNA gene sequences showed that strain JA124(T) clusters with species of the genus Marichromatium belonging to the class Gammaproteobacteria. The highest sequence similarities of strain JA124(T) were found with the type strains of Marichromatium indicum (98 %), Marichromatium purpuratum (95 %) and Marichromatium gracile (93 %). However, DNA-DNA hybridization with Marichromatium indicum DSM 15907(T) revealed relatedness of only 65 % with strain JA124(T). The DNA base composition of strain JA124(T) was 67 mol% G+C (by HPLC). Based on 16S rRNA gene sequence analysis, morphological and physiological characteristics and DNA-DNA hybridization studies, strain JA124(T) (=ATCC BAA-1316(T)=JCM 13911(T)) is sufficiently different from other Marichromatium species to merit its description as the type strain of a novel species, Marichromatium bheemlicum sp. nov

Thioalkalicoccus limnaeus gen. nov., sp. nov., a new alkaliphilic purple sulfur bacterium with bacteriochlorophyll b

Four strains of purple sulfur bacteria containing bacteriochlorophyll b were isolated from cyanobacterial mats of soda lakes in the steppe of south-east Siberia, Russia. Cells of all strains were cocci without gas vesicles. Eventually, cells with flagella were seen in the electron microscope, but motile cells were observed very rarely in cultures. Internal photosynthetic membranes were of the tubular type. Photosynthetic pigments were bacteriochlorophyll b and carotenoids with spectral characteristics similar to 3,4,3',4'-tetrahydrospirilloxanthin. The bacteria were obligately phototrophic and strictly anaerobic. Hydrogen sulfide and elemental sulfur were used as photosynthetic electron donors. Thiosulfate was not used. During growth on sulfide, sulfur globules were formed as intermediate oxidation products, deposited inside the cells and centrally located. In the presence of sulfide and sodium bicarbonate, acetate, malate, propionate, pyruvate, succinate, fumarate and yeast extract were photoassimilated. Growth factors were not required. The new bacterium is an obligate alkaliphile growing at pH 8-10 with an optimum at pH 9. It showed good growth up to 6.0% sodium chloride and up to 8.5% sodium carbonates. Phenotypically, it is similar to Thiococcus pfennigii, but different by virtue of its alkaliphily and salt tolerance. The DNA G+C content was 63.6-64.8 mol %, compared to 69.4-69.9 mol % for Thiococcus pfennigii. The 16S rDNA sequence of strain A26T was approximately 92% similar to that of Thiococcus pfennigii DSM 226 and therefore a new genus and species name, Thioalkalicoccus limnaeus gen. nov. and sp. nov., are proposed for the new bacteriu

Rhodovulum marinum sp. nov., a novel phototrophic purple non-sulfur alphaproteobacterium from marine tides of Visakhapatnam, India

A yellowish-brown bacterium was isolated from enrichment cultures inoculated with seawater samples from the eastern coast of India (Visakhapatnam) under photoheterotrophic conditions. Enrichment and isolation in a medium containing 2 % NaCl (w/v) yielded strain JA128T, which has ovoid to rod-shaped cells, also forms chains and is non-motile. Phylogenetic analysis on the basis of 16S rRNA gene sequences showed that strain JA128T clusters with the Alphaproteobacteria and the sequence similarity with its closest relatives, Rhodovulum iodosum and Rhodovulum sulfidophilum, was 95 %. Strain JA128T contained vesicular intracytoplasmic membranes, bacteriochlorophyll a and carotenoids of the spheroidene series. Strain JA128T was mesophilic, slightly acidophilic, slightly halophilic and grew photoheterotrophically with a number of organic compounds as carbon source and electron donor. It was unable to grow photoautotrophically, chemoautotrophically or by fermentative modes. It did not utilize sulfide, thiosulfate or hydrogen as electron donors. Thiamine was required as a growth factor. Based on the 16S rRNA gene sequence analysis, morphological and physiological characteristics, strain JA128T was significantly different from other species of the genus Rhodovulum and was recognized as a novel species for which the name Rhodovulum marinum sp. nov. is proposed. The type strain is JA128T (=ATCC BAA 1215T=CCUG 52183T=JCM 13300T)

Thiorhodospira sibirica gen. nov., and sp. nov., a new alkaliphilic purple sulfur bacterium from a Siberian soda lake

A new purple sulfur bacterium was isolated from microbial films on decaying plant mass in the near-shore area of the soda lake Malyi Kasytui (pH 9.5, 0.2% salinity) located in the steppe of the Chita region of south-east Siberia. Single cells were vibrioid- or spiral-shaped (3-4 microns wide and 7-20 microns long) and motile by means of a polar tuft of flagella. Internal photosynthetic membranes were of the lamellar type. Lamellae almost filled the whole cell, forming strands and coils. Photosynthetic pigments were bacteriochlorophyll a and carotenoids of the spirilloxanthin group. The new bacterium was strictly anaerobic. Under anoxic conditions, hydrogen sulfide and elemental sulfur were used as photosynthetic electron donors. During growth on sulfide, sulfur globules were formed as intermediate oxidation products. They were deposited outside the cytoplasm of the cells, in the peripheral periplasmic space and extracellularly. Thiosulfate was not used. Carbon dioxide, acetate, pyruvate, propionate, succinate, fumarate and malate were utilized as carbon sources. Optimum growth rates were obtained at pH 9.0 and optimum temperature was 30 degrees C. Good growth was observed in a mineral salts medium containing 5 g sodium bicarbonate l-1 without sodium chloride. The new bacterium tolerated up to 60 g sodium chloride l-1 and up to 80 g sodium carbonates l-1. Growth factors were not required. The DNA G + C composition was 56.0-57.4 mol%. Based on physiological, biochemical and genetic characteristics, the newly isolated bacterium is recognized as a new species of a new genus with the proposed name Thiorhodospira sibirica

Ectothiorhodospira variabilis, sp. nov., an alkaliphilic and halophilic purple sulfur bacterium from soda lakes

During studies of moderately halophilic strains of Ectothiorhodospira from steppe soda lakes, we found a novel group of bacteria related to Ectothiorhodospira haloalkaliphila with salt optima at 50–80 g NaCl l”1. Phylogenetic analysis using 16S rRNA gene sequences of strains from soda lakes in Mongolia, Egypt and Siberia revealed separation of the group of new isolates from other Ectothiorhodospira species, including the closely related Ect. haloalkaliphila. DNA–DNA hybridization studies demonstrated that the new isolates form a homogeneous group at the species level, but at the same time are distinct from related species such as Ect. haloalkaliphila, Ect. vacuolata, Ect. shaposhnikovii and Ect. marina. The new isolates are considered to be strains of a novel species, for which the name Ectothiorhodospira variabilis sp. nov. is proposed, with the type strain WN22T (5VKM B-2479T 5DSM 21381T). Photosynthetic pigments of the novel species are bacteriochlorophyll a and carotenoids of the spirilloxanthin series with spirilloxanthin and derivatives thereof, together with small amounts of lycopene and rhodopin. Gas vesicles are formed by most of the strains, particularly in media containing yeast extract (0.5 g l”1) and acetate (0.5–2.0 g l”1). Sequence analysis of nifH (nitrogenase) and cbbL (RuBisCO) confirmed the assignment of the strains to the genus Ectothiorhodospira and in particular the close relationship to Ect. haloalkaliphila. The novel species Ect. variabilis is found in soda lakes separated by great geographical distances and is an alkaliphilic and halophilic bacterium that tolerates salt concentrations up to 150–200 g NaCl l”1

Rhodobacter johrii sp. nov., an endospore-producing cryptic species isolated from semi-arid tropical soils

An oval to rod shaped phototrophic purple nonsulfur bacterium, strain JA192T was isolated from an enrichment culture of a pasteurized rhizosphere soil sample of jowar crop collected from Godumakunta village, near Hyderabad, India. Strain JA192T is Gram-negative, motile and produces endospores. Phylogenetic analysis on the basis of 16S rRNA gene sequences showed that the strain JA192T is closely related to Rhodobacter sphaeroides DSM 158T (99.9% sequence similarity), Rhodobacter megalophilus JA194T (99.8%) and Rhodobacter azotoformans JCM 9340T (98.1%) and clusters with other species of the genus Rhodobacter of the family Rhodobacteraceae. However, DNA-DNA hybridization with Rba. sphaeroides DSM 158T, Rba. megalophilus JA194T and Rba. azotoformans JCM 9320T showed a relatedness of only 38-57% with respect to JA192T. On the basis of 16S rRNA gene sequence analysis, DNA-DNA hybridization data, morphological, physiological and chemotaxanomic characters, strain JA192T represents a novel species of the genus Rhodobacter, for which the name Rhodobacter johrii sp. nov. is proposed. The type strain is JA192T (= DSM 18678T = JCM 14543T = MTCC 8172T)

Rhodovulum visakhapatnamense sp. nov.

A Gram-negative, rod-shaped, phototrophic bacterium (JA181T) was isolated from a tidal water sample. On the basis of 16S rRNA gene sequence similarity, strain JA181T was shown to belong to the class Alphaproteobacteria, most closely related to Rhodovulum sulfidophilum (97.8 % similarity to the type strain), Rhodovulum adriaticum (93 %), Rhodovulum robiginosum (93 %), Rhodovulum iodosum (94 %), Rhodovulum imhoffii (94 %), Rhodovulum strictum (95 %), Rhodovulum euryhalinum (94.6 %) and Rhodovulum marinum (94.6 %). DNA–DNA hybridization with Rdv. sulfidophilum DSM 1374T (relatedness of 39 % with strain JA181T) and physiological and biochemical tests allowed genotypic and phenotypic differentiation of strain JA181T from the eight Rhodovulum species with validly published names. Strain JA181T therefore represents a novel species, for which the name Rhodovulum visakhapatnamense sp. nov. is proposed (type strain JA181T =JCM 13531T =ATCC BAA-1274T =DSM 17937T)