Phylogenetic relationship of phototrophic purple sulfur bacteria according to pufL and pufM genes

The phylogenetic relationship of purple sulfur bacteria (PSB), of the order Chromatiales (class Gammaproteobacteria), was analyzed based on photosynthetic gene sequences of the pufL and pufM genes, and the results compared to phylogenetic trees and groupings of the 16S rRNA gene. Primers for pufL and pufM genes were constructed and successfully used to amplify the pufLM genes of members of 16 genera of Chromatiales. In total, pufLM and 16S rRNA gene sequences of 66 PSB strains were analyzed, including 29 type strains and 28 new isolates. The inferred phylogenetic trees of the pufLM and 16S rRNA genes reflected a largely similar phylogenetic development suggesting coevolution of these essential genes within the PSB. It is concluded that horizontal gene transfer of pufLM genes within the PSB is highly unlikely, in contrast to the Situation in other groups of anoxygenic phototrophic bacteria belonging to Alpha- and Betaproteobacteria. The phylogeny of pufLM is therefore in good agreement with the current taxonomic classification of PSB. A phylogenetic classification of PSB to the genus level is possible based on their pufL or pufM sequences, and in many cases even to the species level. In addition, our data support a correlation between Puf protein structure and the type of internal photosynthetic membranes (vesicular, lamellar, or tubular

Phylogenetic relationship of phototrophic purple sulfur bacteria according to pufL and pufM genes

The phylogenetic relationship of purple sulfur bacteria (PSB), of the order Chromatiales (class Gammaproteobacteria), was analyzed based on photosynthetic gene sequences of the pufL and pufM genes, and the results compared to phylogenetic trees and groupings of the 16S rRNA gene. Primers for pufL and pufM genes were constructed and successfully used to amplify the pufLM genes of members of 16 genera of Chromatiales. In total, pufLM and 16S rRNA gene sequences of 66 PSB strains were analyzed, including 29 type strains and 28 new isolates. The inferred phylogenetic trees of the pufLM and 16S rRNA genes reflected a largely similar phylogenetic development suggesting coevolution of these essential genes within the PSB. It is concluded that horizontal gene transfer of pufLM genes within the PSB is highly unlikely, in contrast to the situation in other groups of anoxygenic phototrophic bacteria belonging to Alpha- and Betaproteobacteria. The phylogeny of pufLM is therefore in good agreement with the current taxonomic classification of PSB. A phylogenetic classification of PSB to the genus level is possible based on their pufL or pufM sequences, and in many cases even to the species level. In addition, our data support a correlation between Puf protein structure and the type of internal photosynthetic membranes (vesicular, lamellar, or tubular). [Int Microbiol 2009; 12(3):175-185

Bacterial isolates from the bryozoan Membranipora membranacea: influence of culture media on isolation and antimicrobial activity

From specimens of the bryozoan Membranipora membranacea collected in the Baltic Sea, bacteria were isolated on four different media, which significantly increased the diversity of the isolated groups. All isolates were classifiedaccording to 16S rRNA gene sequence analysis and tested for antimicrobial properties using a panel of five indicator strainsand six different media. Each medium featured a unique set of isolated phylotypes, and a phylogenetically diverse collection of isolates was obtained. A total of 96 isolates were assigned to 49 phylotypes and 29 genera. Only one-third of the members of these genera had been isolated previously from comparable sources. The isolates were affiliated with Alpha- and Gammaproteobacteria, Bacilli, and Actinobacteria. A comparable large portion of up to 22 isolates, i.e., 15 phylotypes, probably represent new species. Likewise, 47 isolates (approximately 50%) displayed antibiotic activities, mostly against grampositive indicator strains. Of the active strains, 63.8 % had antibiotic traits only on one or two of the growth media, whereas only 12.7 % inhibited growth on five or all six media. The application of six different media for antimicrobial testing resulted in twice the number of positive hits as obtained with only a single medium. The use of different media for the isolation of bacteria as well as the variation of media considered suitable for the production of antibiotic substances significantly enhanced both the number of isolates obtained and the proportion of antibiotic active cultures. Thus the approach described herein offers an improved strategy in the search for new antibiotic compounds. [Int Microbiol 2012; 15(1):17-32

Bacterial isolates from the bryozoan Membranipora membranacea: influence of culture media on isolation and antimicrobioal activity

From specimens of the bryozoan Membranipora membranacea collected in the Baltic Sea, bacteria were isolated on four different media, which significantly increased the diversity of the isolated groups. All isolates were classifiedaccording to 16S rRNA gene sequence analysis and tested for antimicrobial properties using a panel of five indicator strainsand six different media. Each medium featured a unique set of isolated phylotypes, and a phylogenetically diverse collection of isolates was obtained. A total of 96 isolates were assigned to 49 phylotypes and 29 genera. Only one-third of the members of these genera had been isolated previously from comparable sources. The isolates were affiliated with Alpha- and Gammaproteobacteria, Bacilli, and Actinobacteria. A comparable large portion of up to 22 isolates, i.e., 15 phylotypes, probably represent new species. Likewise, 47 isolates (approximately 50%) displayed antibiotic activities, mostly against grampositive indicator strains. Of the active strains, 63.8 % had antibiotic traits only on one or two of the growth media, whereas only 12.7 % inhibited growth on five or all six media. The application of six different media for antimicrobial testing resulted in twice the number of positive hits as obtained with only a single medium. The use of different media for the isolation of bacteria as well as the variation of media considered suitable for the production of antibiotic substances significantly enhanced both the number of isolates obtained and the proportion of antibiotic active cultures. Thus the approach described herein offers an improved strategy in the search for new antibiotic compound

Kiloniella laminariae, gen. nov., sp. nov., a new alphaproteobacterium from the marine macroalga Laminaria saccharina

A novel alphaproteobacterium, strain LD81(T), was isolated from the marine macroalga. Laminaria saccharina. The bacterium is mesophilic and shows a typical marine growth response. It is a chemoheterotrophic aerobe with the potential for denitrification. Growth optima are 25 degrees C, pH 5.5 and 3% NaCl. Strain LD81(T) has a unique phylogenetic position, not fitting any of the known families of the Alphaproteobacteria. The 16S rRNA gene sequence revealed a distant relationship to species of several orders of the Alphaproteobacteria, with less than 90% sequence similarity. Phylogenetically, strain LD81(T) is related to the type strains of Terasakiella pusilla (88.4% 16S rRNA gene sequence similarity) and the three Thalassospira species (88.9-89.2%. It forms a cluster with these bacteria and a novel as-yet undescribed isolate (KOPRI 13522; 96.6% sequence similarity). Strain LD81(T) has a relatively low DNA G + C content (51.1 mol%) and, due to its distant phylogenetic position from all other alphaprotecibacteria, strain LD81(T) (=NCIMB 14374(T) =JCM 14845(T)) is considered as the type strain of a novel species within a new genus, for which the name Kiloniella laminariae gen. nov., sp. nov. is proposed. The genus Kiloniella represents the type of the new family Kiloniellaceae fam. nov. and order Kiloniellales ord. nov

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)

Thiohalocapsa marina sp. nov., from an Indian marine aquaculture pond

A spherical-shaped, phototrophic, purple sulfur bacterium was isolated in pure culture from anoxic sediment in a marine aquaculture pond near Bheemli (India). Strain JA142T is Gram-negative and non-motile. It has a requirement for NaCl (optimum of 2 % and maximum of 6 % w/v NaCl). Intracellular photosynthetic membranes are of the vesicular type. In vivo absorption spectra indicate the presence of bacteriochlorophyll a and carotenoids of the okenone series as photosynthetic pigments. Phylogenetic analysis on the basis of 16S rRNA gene sequences showed that strain JA142T is related to halophilic purple sulfur bacteria of the genera Thiohalocapsa and Halochromatium, with the highest sequence similarity to Thiohalocapsa halophila DSM 6210T (97.5 %). Morphological and physiological characteristics differentiate strain JA142T from other species of the genera Halochromatium and Thiohalocapsa. Strain JA142T is sufficiently different from Thiohalocapsa halophila based on 16S rRNA gene sequence analysis and morphological and physiological characteristics to allow the proposal of a novel species, Thiohalocapsa marina sp. nov., with the type strain JA142T (=JCM 14780T =DSM 19078T). The GenBank/EMBL/DDBJ accession number for the 16S rRNA gene sequence of strain JA142T is AM491592. A phase-contrast micrograph of cells of strain JA142T, whole-cell and acetone absorption spectra and 16S rRNA gene sequence-based neighbour-joining, maximum-likelihood, minimum-evolution and maximum-parsimony trees are available as supplementary material with the online version of this paper