Vesicular Intracytoplasmic Membrane Research Articles

Rhodobacter flagellatus sp. nov., a thermophilic bacterium isolated from a hot spring.

A thermophilic bacterium, designated SYSU G03088T, was isolated from Moincer hot spring, Tibet, PR China. Polyphasic taxonomic analyses and whole-genome sequencing were used to determine the taxonomic position and genomic profiles of the strain. Phylogenetic analysis using 16S rRNA gene sequence indicated that SYSU G03088T showed highest sequence similarity to Rhodobacter blasticus CGMCC 1.3365T (96.0 % sequence identity). The strain could be differentiated from most recognized species of the genus Rhodobacter by its slightly purple colony colour, distinct phenotypic characters and low ANI values. Cells were Gram-staining negative, and oval-to-rod shaped. Poly-β-hydroxybutyrate and vesicular intracytoplasmic membrane structures were formed inside cells. Growth occurred optimally at 45 °C and pH 7.0. Ubiquinone 10 was the only respiratory quinone. The major fatty acids (>10 %) were C18 : 0, C18 : 1ω7c 11-methyl and summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c). The detected polar lipids of SYSU G03088T included diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol and phosphatidylmethylethanolamine. The DNA G+C content of SYSU G03088T was 67.7 % (genome). On the basis of the differences in the phenotypic characteristics and phylogenetic analyses, SYSU G03088T is considered to represent a novel species of the genus Rhodobacter, for which the name Rhodobacter flagellatus sp. nov. is proposed. The type strain is SYSU G03088T (=CGMCC 1.16876T=KCTC 72354T).

Descriptions of Roseiarcus fermentans gen. nov., sp. nov., a bacteriochlorophyll a-containing fermentative bacterium related phylogenetically to alphaproteobacterial methanotrophs, and of the family Roseiarcaceae fam. nov.

A light-pink-pigmented, microaerophilic bacterium was obtained from a methanotrophic consortium enriched from acidic Sphagnum peat and designated strain Pf56(T). Cells of this bacterium were Gram-negative, non-motile, thick curved rods that contained a vesicular intracytoplasmic membrane system characteristic of some purple non-sulfur alphaproteobacteria. The absorption spectrum of acetone/methanol extracts of cells grown in the light showed maxima at 363, 475, 505, 601 and 770 nm; the peaks at 363 and 770 nm are characteristic of bacteriochlorophyll a. However, in contrast to purple non-sulfur bacteria, strain Pf56(T) was unable to grow phototrophically under anoxic conditions in the light. Best growth occurred on some sugars and organic acids under micro-oxic conditions by means of fermentation. The fermentation products were propionate, acetate and hydrogen. Slow chemo-organotrophic growth was also observed under fully oxic conditions. Light stimulated growth. C1 substrates were not utilized. Strain Pf56(T) grew at pH 4.0-7.0 (optimum pH 5.5-6.5) and at 15-30 °C (optimum 22-28 °C). The major cellular fatty acids were 19 : 0 cyclo ω8c and 18 : 1ω7c; quinones were represented by ubiquinone Q-10. The G+C content of the DNA was 70.0 mol%. Strain Pf56 displays 93.6-94.7 and 92.7-93.7% 16S rRNA gene sequence similarity to members of the families Methylocystaceae and Beijerinckiaceae, respectively, and belongs to a large cluster of environmental sequences retrieved from various wetlands and forest soils in cultivation-independent studies. Phenotypic, genotypic and chemotaxonomic characteristics of strain Pf56(T) suggest that it represents a novel genus and species of bacteriochlorophyll a-containing fermentative bacteria, for which the name Roseiarcus fermentans gen. nov., sp. nov. is proposed. Strain Pf56(T) ( = DSM 24875(T) = VKM B-2876(T)) is the type strain of Roseiarcus fermentans, and is also the first characterized member of a novel family within the class Alphaproteobacteria, Roseiarcaceae fam. nov.

Rhodovastum atsumiense gen. nov., sp. nov., a phototrophic alphaproteobacterium isolated from paddy soil

A photoorganotrophic alphaproteobacterium designated strain G2-11(T) was isolated from submerged paddy soil. This bacterium had relatively large, oval to rod-shaped cells (2.0-3.0x3.0-10 microm). Cells were motile by means of single polar flagella. The color of phototrophically growing cultures was reddish-brown. The cell extract had absorption maxima at 375, 465, 492, 529, 592, 804, and 844 nm, indicating the presence of bacteriochlorophyll a and carotenoides of the spirilloxanthin series. Vesicular intracytoplasmic membranes were present. The main component of cellular fatty acids was C(18:1)omega7c. Ubiquinone-10 and rhodoquinone-10 were the major quinones. A 16S rRNA gene sequence analysis revealed that the isolate is closest to the acidophilic aerobic photosynthetic bacterium Acidisphaera rubrifaciens strain HS-AP3(T) (93.3% similarity). The G+C content of genomic DNA is 67.8 mol%. The name Rhodovastum atsumiense gen. nov., sp. nov. is proposed for the novel isolate. The type strain is strain G2-11(T) (=NBRC 104268(T)=KCTC 5708(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)).

Halochromatium roseum sp. nov., a non-motile phototrophic gammaproteobacterium with gas vesicles, and emended description of the genus Halochromatium

A rod-shaped, marine, phototrophic, purple sulfur bacterium containing gas vesicles was isolated from a marine solar saltern at Kakinada, India. Cells of strain JA134(T) are Gram-negative, non-motile rods, with vesicular intracytoplasmic membranes, and the strain has an absolute requirement for NaCl for growth. Bacteriochlorophyll a and the carotenoid okenone are present as photosynthetic pigments. Phylogenetic analysis on the basis of 16S rRNA gene sequences showed that strain JA134(T) clusters with members of the genus Halochromatium, but is distinctly separated from the species reported so far. The morphological and physiological differences of strain JA134(T) from other Halochromatium species support the description of strain JA134(T) (=ATCC BAA-1363(T) =DSM 18859(T) =JCM 14151(T)) as the type strain of a novel species, Halochromatium roseum sp. nov.

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 JA128(T), 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 JA128(T) clusters with the Alphaproteobacteria and the sequence similarity with its closest relatives, Rhodovulum iodosum and Rhodovulum sulfidophilum, was 95 %. Strain JA128(T) contained vesicular intracytoplasmic membranes, bacteriochlorophyll a and carotenoids of the spheroidene series. Strain JA128(T) 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 JA128(T) 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 JA128(T) (=ATCC BAA 1215(T)=CCUG 52183(T)=JCM 13300(T)).

Rhodobaca bogoriensis gen. nov. and sp. nov., an alkaliphilic purple nonsulfur bacterium from African Rift Valley soda lakes.

From enrichment cultures established for purple nonsulfur bacteria using water and sediment samples from Lake Bogoria and Crater Lake, two soda lakes in the African Rift Valley, three strains of purple nonsulfur bacteria were isolated; strain LBB1 was studied in detail. Cells of strain LBB1 were motile and spherical to rod-shaped, suggesting a relationship to Rhodobacter or Rhodovulum species, and the organism was capable of both phototrophic and chemotrophic growth on a wide variety of organic compounds. Phototrophically grown cultures were yellow to yellow-brown in color and grew optimally at pH 9 (pH range 7.5-10) and 1% NaCl (range 0-10%). In physiological studies of strain LBB1, neither photoautotrophy (H2- or sulfide-dependent) nor nitrogen fixation was observed. Absorption spectra revealed that all three strains contained bacteriochlorophyll a and carotenoids of the spheroidene pathway and synthesized only a light-harvesting (LH) I-type photosynthetic antenna complex. Electron microscopy of cells of strain LBB1 revealed a vesicular intracytoplasmic membrane system, although only a few vesicles were observed per cell. The G+C content of strain LBB1 DNA was 59 mol%, significantly lower than that of known Rhodobacter and Rhodovulum species, and its phylogeny as determined by ribosomal RNA gene sequencing placed it within the Rhodobacter/Rhodovulum clade yet distinct from all described species of either of these genera. The unique assemblage of properties observed in strain LBB1 warrants its inclusion in a new genus of purple nonsulfur bacteria and the name Rhodobaca bogoriensis is proposed herein, the genus name reflecting morphological characteristics and the species epithet referring to the habitat.

Isolation and Characterization of Rhodovulum strictum sp. nov. and Some Other Purple Nonsulfur Bacteria from Colored Blooms in Tidal and Seawater Pools

Several strains of phototrophic purple nonsulfur bacteria were isolated from colored blooms occurring in tidal and seawater pools in Japan. All of these isolates had ovoid to rod-shaped cells that were motile by means of single polar flagella and contained vesicular intracytoplasmic membranes together with bacteriochlorophyll a and carotenoids of the spheroidene series. They produced ubiquinone 10 as the major quinone and contained straight-chain fatty acids, with C18:1 predominating. They were mesophilic, halophilic, and photoheterotrophic, utilized sulfide and thiosulfate as electron donors for phototrophic growth, and photoassimilated a wide variety of organic compounds as carbon sources. Our results suggested that all of these isolates are members of the recently described genus Rhodovulum. The isolates were classified into four groups (designated groups I through IV) on the basis of phenotypic and genotypic data. The group I isolates, which were the most abundant purple nonsulfur bacteria recovered from the blooms, grew in the presence of NaCl concentrations ranging from 0.5 to 3.0% (optimum NaCl concentration, 0.8%) and at pH values ranging from 7.5 to 9.0 (optimum pH, 8.0 to 8.5). On the basis of these unique physiological traits, together with genotypic and phylogenetic data, we propose that the group I isolates should be classified as members of a new species, Rhodovulum strictum. The group II isolates were identified definitely as Rhodovulum sulfidophilum, and the group III and IV isolates were phenotypically most similar to R. sulfidophilum and Rhodovulum adriaticum, respectively, but could be differentiated from these species by DNA-DNA pairing data.

Rearrangement of the Species and Genera of the Phototrophic "Purple Nonsulfur Bacteria"

A rearrangement of the species of the “purple nonsulfur bacteria” (Rhodospirillaceae) is proposed. The species with vesicular intracytoplasmic membranes are removed from the genus Rhodopseudomonas and are placed in the new genera Rhodobacter and Rhodopila as Rhodobacter capsulatus (type species), Rhodobacter sphaeroides, Rhodobacter sulfidophilus, Rhodobacter adriaticus, and Rhodopila globiformis (type species). Rhodopseudomonas gelatinosa and Rhodospirillum tenue are united with Rhodocyclus purpureus (type species) in the genus Rhodocyclus as Rhodocyclus gelatinosus and Rhodocyclus tenuis. The decisions for this rearrangement were based mainly upon morphological and physiological properties; supporting arguments were based on structural similarities of macromolecular cell constituents.

Development and growth of photosynthetic membranes of Rhodospirillum rubrum.

In cell-free extracts from low-aeration suspensions of Rhodospirillum rubrum strain G-9, bacteriochlorophyll a was distributed in two bands after rate-zone sedimentation in sucrose density gradients. From the physicochemical properties of these fractions, it was concluded that the upper band consisted of small membrane fragments, whereas the major band was composed of fragmented vesicular intracytoplasmic membrane (chromatophores). After a pulse with L-[35S]methionine, apparent polypeptide subunits of the reaction center and light-harvesting complexes within the upper pigmented fraction were labeled more rapidly than those of chromatophores; after a chase with excess unlabeled L-methionine, radioactivity from these components within the upper band appeared to be chased into the corresponding polypeptides of chromatophores. These labeling patterns are interpreted to reflect growth initiation and maturation of the photosynthetic apparatus and may, in part, represent a general mechanism for the development of vesicular intracytoplasmic membranes.

The influence of 2-hydroxybiphenyl on membranes of Rhodospirillum rubrum.

The effects of 2-hydroxybiphenyl upon intracytoplasmic membranes of Rhodospirillum rubrum were investigated. At concentrations of 110 and 165 μM of 2-hydroxybiphenyl growth was delayed, it stopped completely at a concentration of 330 μM. In the latter case, instead of vesicular intracytoplasmic membranes concentric membraneous layers were found in electronmicrographs of whole cells. Inhibitor concentrations which still permit growth do not change the general appearance of intracytoplasmic membranes either in situ or in the isolated form. However, the formation of intracytoplasmic membranes was more affected by the presence of the inhibitor than was growth. Although by electron microscopy no effect of 2-hydroxybiphenyl on intracytoplasmic membranes could be revealed there were considerable influences on membrane composition. This concerned the formation of colored carotenoids and specifically the patterns of membrane proteins.

The intracytoplasmic membrane system of the bacteroids of subterraneum clover nodules (Trifolium subterraneum L.)

A vesicular intracytoplasmic membrane system is demonstrated in bacteroids from the leghaemoglobin filled zone of effective Trifolium subterraneum nodules after KMnO4 and OsO4 fixation. The system appears to be present in all mature bacteroids from this zone, and is derived from tubular invaginations of the plasma membrane of the bacteriod. A granular substance similar to the bacteroid cytoplasm is found in the vesicles which are bounded by a tripartite membrane approximately 80 A wide, while the interspace between the vesicles is filled with a material of similar appearance to that in the interspace between bacteroid plasma membrane and cell wall.