Specific detection of green sulfur bacteria by in situ hybridization with a fluorescently labeled oligonucleotide probe

1999 ◽  
Vol 171 (4) ◽  
pp. 265-272 ◽  
Author(s):  
Christian Tuschak ◽  
Jens Glaeser ◽  
J. Overmann
Keyword(s):  
In Situ Hybridization ◽  
Oligonucleotide Probe ◽  
Green Sulfur Bacteria ◽  
Specific Detection ◽  
Sulfur Bacteria ◽  
Green Sulfur

scholarly journals Physiology and Phylogeny of Green Sulfur Bacteria Forming a Monospecific Phototrophic Assemblage at a Depth of 100 Meters in the Black Sea

2005 ◽  
Vol 71 (12) ◽  
pp. 8049-8060 ◽  
Author(s):  
Ann K. Manske ◽  
Jens Glaeser ◽  
Marcel M. M. Kuypers ◽  
Jörg Overmann
Keyword(s):  
Black Sea ◽  
Sulfide Oxidation ◽  
Green Sulfur Bacteria ◽  
The Black Sea ◽  
Rrna Gene ◽  
Green Sulfur Bacterium ◽  
Sulfur Bacteria ◽  
Light Intensities ◽  
Green Sulfur

ABSTRACT The biomass, phylogenetic composition, and photoautotrophic metabolism of green sulfur bacteria in the Black Sea was assessed in situ and in laboratory enrichments. In the center of the western basin, bacteriochlorophyll e (BChl e) was detected between depths of 90 and 120 m and reached maxima of 54 and 68 ng liter−1. High-pressure liquid chromatography analysis revealed a dominance of farnesyl esters and the presence of four unusual geranyl ester homologs of BChl e. Only traces of BChl e (8 ng liter−1) were found at the northwestern slope of the Black Sea basin, where the chemocline was positioned at a significantly greater depth of 140 m. Stable carbon isotope fractionation values of farnesol indicated an autotrophic growth mode of the green sulfur bacteria. For the first time, light intensities in the Black Sea chemocline were determined employing an integrating quantum meter, which yielded maximum values between 0.0022 and 0.00075 μmol quanta m−2 s−1 at the top of the green sulfur bacterial layer around solar noon in December. These values represent by far the lowest values reported for any habitat of photosynthetic organisms. Only one 16S rRNA gene sequence type was detected in the chemocline using PCR primers specific for green sulfur bacteria. This previously unknown phylotype groups with the marine cluster of the Chlorobiaceae and was successfully enriched in a mineral medium containing sulfide, dithionite, and freshly prepared yeast extract. Under precisely controlled laboratory conditions, the enriched green sulfur bacterium proved to be capable of exploiting light intensities as low as 0.015 μmol quanta m−2 s−1 for photosynthetic 14CO2 fixation. Calculated in situ doubling times of the green sulfur bacterium range between 3.1 and 26 years depending on the season, and anoxygenic photosynthesis contributes only 0.002 to 0.01% to total sulfide oxidation in the chemocline. The stable population of green sulfur bacteria in the Black Sea chemocline thus represents the most extremely low-light-adapted and slowest-growing type of phototroph known to date.

Specific Detection of Legionella pneumophila: Construction of a New 16S rRNA-Targeted Oligonucleotide Probe

1998 ◽  
Vol 64 (7) ◽  
pp. 2686-2690 ◽  
Author(s):  
Dorothee Grimm ◽  
Hilde Merkert ◽  
Wolfgang Ludwig ◽  
Karl-Heinz Schleifer ◽  
Jörg Hacker ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
16S Rrna ◽  
Legionella Pneumophila ◽  
Oligonucleotide Probe ◽  
Acanthamoeba Castellanii ◽  
Comparative Sequence Analysis ◽  
Specific Detection ◽  
Reference Strains ◽  
Legionella Micdadei

Based on comparative sequence analysis, we have designed an oligonucleotide probe complementary to a region of 16S rRNA ofLegionella pneumophila which allows the differentiation ofL. pneumophila from other Legionellaspecies without cultivation. The specificity of the new probe, LEGPNE1, was tested by in situ hybridization to a total of four serogroups of six strains of L. pneumophila, five differentLegionella spp. and three nonlegionella species as reference strains. Furthermore, L. pneumophilacells could be easily distinguished from Legionella micdadei and Pseudomonas aeruginosa cells by using in situ hybridization with probes LEGPNE1, LEG705, and EUB338 after infection of the protozoan Acanthamoeba castellanii.

scholarly journals Characterization and In Situ Carbon Metabolism of Phototrophic Consortia

2003 ◽  
Vol 69 (7) ◽  
pp. 3739-3750 ◽  
Author(s):  
Jens Glaeser ◽  
Jörg Overmann
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Stable Carbon Isotope ◽  
Green Sulfur Bacteria ◽  
Purple Sulfur Bacteria ◽  
Rrna Gene ◽  
Sulfur Bacteria ◽  
Green Sulfur

ABSTRACT A dense population of the phototrophic consortium “Pelochromatium roseum” was investigated in the chemocline of a temperate holomictic lake (Lake Dagow, Brandenburg, Germany). Fluorescence in situ hybridization revealed that the brown epibionts of “P. roseum” constituted up to 37% of the total bacterial cell number and up to 88% of all green sulfur bacteria present in the chemocline. Specific amplification of 16S rRNA gene fragments of green sulfur bacteria and denaturing gradient gel electrophoresis fingerprinting yielded a maximum of four different DNA bands depending on the year of study, indicating that the diversity of green sulfur bacteria was low. The 465-bp 16S rRNA gene sequence of the epibiont of “P. roseum” was obtained after sorting of individual consortia by micromanipulation, followed by a highly sensitive PCR. The sequence obtained represents a new phylotype within the radiation of green sulfur bacteria. Maximum light-dependent H14CO3 − fixation in the chemocline in the presence of 3-(3,4-dichlorophenyl)-1,1-dimethylurea suggested that there was anaerobic autotrophic growth of the green sulfur bacteria. The metabolism of the epibionts was further studied by determining stable carbon isotope ratios (δ13C) of their specific biomarkers. Analysis of photosynthetic pigments by high-performance liquid chromatography revealed the presence of high concentrations of bacteriochlorophyll (BChl) e and smaller amounts of BChl a and d and chlorophyll a in the chemocline. Unexpectedly, isorenieratene and β-isorenieratene, carotenoids typical of other brown members of the green sulfur bacteria, were absent. Instead, four different esterifying alcohols of BChl e were isolated as biomarkers of green sulfur bacterial epibionts, and their δ13C values were determined. Farnesol, tetradecanol, hexadecanol, and hexadecenol all were significantly enriched in 13C compared to bulk dissolved and particulate organic carbon and compared to the biomarkers of purple sulfur bacteria. The difference between the δ13C values of farnesol, the major esterifying alcohol of BChl e, and CO2 was −7.1%, which provides clear evidence that the mode of growth of the green sulfur bacterial epibionts of “P. roseum” in situ is photoautotrophic.

scholarly journals Expression and localization of the mRNA for DNA (cytosine-5)- methyltransferase in mouse seminiferous tubules.

1994 ◽  
Vol 42 (9) ◽  
pp. 1271-1276 ◽  
Author(s):  
M Numata ◽  
T Ono ◽  
S Iseki
Keyword(s):  
In Situ Hybridization ◽  
Significant Role ◽  
Meiotic Prophase ◽  
Oligonucleotide Probe ◽  
Mouse Testis ◽  
Seminiferous Tubules ◽  
Hybridization Histochemistry ◽  
In Situ Hybridization Histochemistry

DNA (cytosine-5)-methyltransferase (DNA MTase) is the only enzyme known to be involved in the methylation of mammalian DNA. Although the expression of DNA MTase gene is abundant in the testis, little is known about the role of this enzyme during spermatogenesis. We examined the distribution of DNA MTase mRNA in mouse testis by in situ hybridization histochemistry with an oligonucleotide probe. The mRNA signal was observed in the seminiferous tubules and was localized predominantly in spermatogonia and spermatocytes, particularly during the earlier steps of meiotic prophase I, with maximal intensity in the early pachytene cells. These results suggest some significant role for DNA MTase in spermatogenesis.

scholarly journals Signature pigments of green sulfur bacteria in lower Pleistocene deposits from the Banyoles lacustrine area (Spain)

2005 ◽  
Vol 34 (2) ◽  
pp. 271-280 ◽  
Author(s):  
N. Mallorquí ◽  
J.B. Arellano ◽  
C.M. Borrego ◽  
L.J. Garcia-Gil
Keyword(s):  
Green Sulfur Bacteria ◽  
Sulfur Bacteria ◽  
Lower Pleistocene ◽  
Green Sulfur

scholarly journals Differential RNA Sequencing Implicates Sulfide as the Master Regulator of S 0 Metabolism in Chlorobaculum tepidum and Other Green Sulfur Bacteria

2017 ◽  
Vol 84 (3) ◽  
Author(s):  
Jacob M. Hilzinger ◽  
Vidhyavathi Raman ◽  
Kevin E. Shuman ◽  
Brian J. Eddie ◽  
Thomas E. Hanson
Keyword(s):  
Gene Expression ◽  
Elemental Sulfur ◽  
Green Sulfur Bacteria ◽  
Electron Donors ◽  
Reduced Sulfur Compounds ◽  
Promoter Elements ◽  
Content Type ◽  
Sulfur Bacteria ◽  
Chlorobaculum Tepidum ◽  
Green Sulfur

ABSTRACT The green sulfur bacteria ( Chlorobiaceae ) are anaerobes that use electrons from reduced sulfur compounds (sulfide, S 0 , and thiosulfate) as electron donors for photoautotrophic growth. Chlorobaculum tepidum , the model system for the Chlorobiaceae , both produces and consumes extracellular S 0 globules depending on the availability of sulfide in the environment. These physiological changes imply significant changes in gene regulation, which has been observed when sulfide is added to Cba. tepidum growing on thiosulfate. However, the underlying mechanisms driving these gene expression changes, i.e., the specific regulators and promoter elements involved, have not yet been defined. Here, differential RNA sequencing (dRNA-seq) was used to globally identify transcript start sites (TSS) that were present during growth on sulfide, biogenic S 0 , and thiosulfate as sole electron donors. TSS positions were used in combination with RNA-seq data from cultures growing on these same electron donors to identify both basal promoter elements and motifs associated with electron donor-dependent transcriptional regulation. These motifs were conserved across homologous Chlorobiaceae promoters. Two lines of evidence suggest that sulfide-mediated repression is the dominant regulatory mode in Cba. tepidum . First, motifs associated with genes regulated by sulfide overlap key basal promoter elements. Second, deletion of the Cba. tepidum 1277 ( CT1277 ) gene, encoding a putative regulatory protein, leads to constitutive overexpression of the sulfide:quinone oxidoreductase CT1087 in the absence of sulfide. The results suggest that sulfide is the master regulator of sulfur metabolism in Cba. tepidum and the Chlorobiaceae . Finally, the identification of basal promoter elements with differing strengths will further the development of synthetic biology in Cba. tepidum and perhaps other Chlorobiaceae . IMPORTANCE Elemental sulfur is a key intermediate in biogeochemical sulfur cycling. The photoautotrophic green sulfur bacterium Chlorobaculum tepidum either produces or consumes elemental sulfur depending on the availability of sulfide in the environment. Our results reveal transcriptional dynamics of Chlorobaculum tepidum on elemental sulfur and increase our understanding of the mechanisms of transcriptional regulation governing growth on different reduced sulfur compounds. This report identifies genes and sequence motifs that likely play significant roles in the production and consumption of elemental sulfur. Beyond this focused impact, this report paves the way for the development of synthetic biology in Chlorobaculum tepidum and other Chlorobiaceae by providing a comprehensive identification of promoter elements for control of gene expression, a key element of strain engineering.

Sign in / Sign up

Export Citation Format

Share Document