scholarly journals Ribulose-1,5-bisphosphate carboxylase/oxygenase genes as a functional marker for chemolithoautotrophic halophilic sulfur-oxidizing bacteria in hypersaline habitats

Microbiology ◽  
2010 ◽  
Vol 156 (7) ◽  
pp. 2016-2025 ◽  
Author(s):  
Tatjana P. Tourova ◽  
Olga L. Kovaleva ◽  
Dimitry Yu. Sorokin ◽  
Gerard Muyzer

The presence and diversity of the cbb genes encoding the large subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) (a key enzyme of the Calvin–Benson cycle of autotrophic CO2 assimilation) were investigated in pure cultures of seven genera of halophilic chemolithoautotrophic sulfur-oxidizing bacteria (SOB) and in sediments from a hypersaline lake in which such bacteria have been recently discovered. All of the halophilic SOB strains (with the exception of Thiohalomonas nitratireducens) possessed the cbbL gene encoding RuBisCO form I, while the cbbM gene encoding RuBisCO form II was detected only in some of the pure cultures. The general topologies of the CbbL/CbbM trees and the 16S rRNA gene tree were different, but both markers showed that the halophilic SOB genera formed independent lineages in the Gammaproteobacteria. In some cases, such as with several strains of the genus Thiohalospira and with Thioalkalibacter halophilus, the cbbL clustering was incongruent with the positions of these strains on the ribosomal tree. In the cbbM tree, the clustering of Thiohalospira and Thiohalorhabdus strains was incongruent with their branching in both cbbL and 16S rRNA gene trees. cbbL and cbbM genes related to those found in the analysed halophilic SOB were also detected in a sediment from a hypersaline lake in Kulunda Steppe (Russia). Most of the cbbL and cbbM genes belonged to members of the genus Thiohalorhabdus. In the cbbL clone library, sequences related to those of Halothiobacillus and Thiohalospira were detected as minor components. Some of the environmental cbbM sequences belonged to as yet unknown phylotypes, representing deep lineages of halophilic autotrophs.

2007 ◽  
Vol 57 (10) ◽  
pp. 2387-2398 ◽  
Author(s):  
Tatjana P. Tourova ◽  
Elizaveta M. Spiridonova ◽  
Ivan A. Berg ◽  
Natalia V. Slobodova ◽  
Eugenia S. Boulygina ◽  
...  

The occurrence of genes encoding nitrogenase and ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO) was investigated in the members of the family Ectothiorhodospiraceae. This family forms a separate phylogenetic lineage within the Gammaproteobacteria according to 16S rRNA gene sequence analysis and mostly includes photo- and chemoautotrophic halophilic and haloalkaliphilic bacteria. The cbbL gene encoding the large subunit of ‘green-like’ form I RubisCO was found in all strains, except the type strains of Alkalispirillum mobile and Arhodomonas aquaeolei. The nifH gene encoding nitrogenase reductase was present in all investigated species of the phototrophic genera Ectothiorhodospira, Halorhodospira and Thiorhodospira, but not of the genus Ectothiorhodosinus. Unexpectedly, nifH fragments were also obtained for the chemotrophic species Thioalkalispira microaerophila and Alkalilimnicola halodurans, for which diazotrophic potential has not previously been assumed. The cbbL-, nifH- and 16S rRNA gene-based trees were not highly congruent in their branching patterns since, in the ‘RubisCO’ and ‘nitrogenase’ trees, representatives of the Ectothiorhodospiraceae are divided in a number of broadly distributed clusters and branches. However, the data obtained may be regarded as evidence of the monophyletic origin of the cbbL and nifH genes in most species within the family Ectothiorhodospiraceae and mainly corresponded to the current taxonomic structure of this family. The cbbL phylogeny of the chemolithoautotrophic sulfur-oxidizers Thioalkalivibrio nitratireducens and Thioalkalivibrio paradoxus and the nitrifier Nitrococcus mobilis deviated significantly from the 16S-rRNA gene-based phylogeny. These species clustered with one of the duplicated cbbL genes of the purple sulfur bacterium Allochromatium vinosum, a member of the family Chromatiaceae.


2006 ◽  
Vol 56 (5) ◽  
pp. 937-945 ◽  
Author(s):  
Bożena M. Korczak ◽  
Regina Stieber ◽  
Stefan Emler ◽  
André P. Burnens ◽  
Joachim Frey ◽  
...  

The genus Campylobacter comprises 17 species, some of which are important animal and human pathogens. To gain more insight into the genetic relatedness of this genus and to improve the molecular tools available for diagnosis, a universal sequencing approach was established for the gene encoding the beta-subunit of RNA polymerase (rpoB) for the genus Campylobacter. A total of 59 strains, including the type strains of currently recognized species as well as field isolates, were investigated in the study. A primer set specific for Campylobacter species enabled straightforward amplification and sequencing of a 530 bp fragment of the rpoB gene. The 16S rRNA gene sequences of all of the strains were determined in parallel. A good congruence was obtained between 16S rRNA and rpoB gene sequence-based trees within the genus Campylobacter. The branching of the rpoB tree was similar to that of the 16S rRNA gene tree, even though a few discrepancies were observed for certain species. The resolution of the rpoB gene within the genus Campylobacter was generally much higher than that of the 16S rRNA gene sequence, resulting in a clear separation of most species and even some subspecies. The universally applicable amplification and sequencing approach for partial rpoB gene sequence determination provides a powerful tool for DNA sequence-based discrimination of Campylobacter species.


2005 ◽  
Vol 71 (4) ◽  
pp. 2106-2112 ◽  
Author(s):  
Chuanlun L. Zhang ◽  
Zhiyong Huang ◽  
James Cantu ◽  
Richard D. Pancost ◽  
Robin L. Brigmon ◽  
...  

ABSTRACT White and orange mats are ubiquitous on surface sediments associated with gas hydrates and cold seeps in the Gulf of Mexico. The goal of this study was to determine the predominant pathways for carbon cycling within an orange mat in Green Canyon (GC) block GC 234 in the Gulf of Mexico. Our approach incorporated laser-scanning confocal microscopy, lipid biomarkers, stable carbon isotopes, and 16S rRNA gene sequencing. Confocal microscopy showed the predominance of filamentous microorganisms (4 to 5 μm in diameter) in the mat sample, which are characteristic of Beggiatoa. The phospholipid fatty acids extracted from the mat sample were dominated by 16:1ω7c/t (67%), 18:1ω7c (17%), and 16:0 (8%), which are consistent with lipid profiles of known sulfur-oxidizing bacteria, including Beggiatoa. These results are supported by the 16S rRNA gene analysis of the mat material, which yielded sequences that are all related to the vacuolated sulfur-oxidizing bacteria, including Beggiatoa, Thioploca, and Thiomargarita. The δ13C value of total biomass was −28.6‰; those of individual fatty acids were −29.4 to −33.7‰. These values suggested heterotrophic growth of Beggiatoa on organic substrates that may have δ13C values characteristic of crude oil or on their by-products from microbial degradation. This study demonstrated that integrating lipid biomarkers, stable isotopes, and molecular DNA could enhance our understanding of the metabolic functions of Beggiatoa mats in sulfide-rich marine sediments associated with gas hydrates in the Gulf of Mexico and other locations.


2015 ◽  
Vol 65 (Pt_6) ◽  
pp. 1786-1793 ◽  
Author(s):  
Wallace Rafael Souza ◽  
Rafael Eduardo Silva ◽  
Michael Goodfellow ◽  
Kanungnid Busarakam ◽  
Fernanda Sales Figueiro ◽  
...  

Strain SB026T was isolated from Brazilian rainforest soil and its taxonomic position established using data from a polyphasic study. The organism showed a combination of chemotaxonomic and morphological features consistent with its classification in the genus Amycolatopsis and formed a branch in the Amycolatopsis 16S rRNA gene tree together with Amycolatopsis bullii NRRL B-24847T, Amycolatopsis plumensis NRRL B-24324T, Amycolatopsis tolypomycina DSM 44544T and Amycolatopsis vancoresmycina NRRL B-24208T. It was related most closely to A. bullii NRRL B-24847T (99.0 % 16S rRNA gene sequence similarity), but was distinguished from this strain by a low level of DNA–DNA relatedness (~46 %) and discriminatory phenotypic properties. Based on the combined genotypic and phenotypic data, it is proposed that the isolate should be classified in the genus Amycolatopsis as representing a novel species, Amycolatopsis rhabdoformis sp. nov. The type strain is SB026T ( = CBMAI 1694T = CMAA 1285T = NCIMB 14900T).


2020 ◽  
Vol 10 (20) ◽  
pp. 7111
Author(s):  
Yolanda Espín ◽  
Giuliana Aranzulla ◽  
Manuel Álvarez-Ortí ◽  
Juan José Gómez-Alday

The chemical composition of groundwater and related surface water ecosystems can be modified by intensive agricultural activities. This is the case of the Natural Reserve of Pétrola saline lake (Albacete, SE Spain), which constitutes the discharge area of an unconfined aquifer. The extended use of fertilizers and pesticides poses a threat to ecosystem sustainability. One of the most applied herbicides worldwide has been atrazine. Despite being prohibited in Spain since 2007 by European regulations, atrazine can still be detected due to its high persistence in the environment. Atrazine degradation pathways are mediated by biological processes performed by microorganisms with adapted metabolic mechanisms that make in situ bioremediation possible. To evaluate the presence of such microorganisms in the unconfined aquifer, groundwater was collected from a flowing 37.9 m deep piezometer. DNA was extracted, and the bacterial 16S rRNA gene was amplified and cloned. Later, 93 clones were sequenced, providing the first molecular assessment of bacterial community structure in the deep zones of the aquifer. Some of these bacteria have been previously described to be involved in atrazine degradation. In addition, 14 bacteria were isolated from the groundwater samples and identified by 16S rRNA gene sequencing. DNA from these bacteria was subjected to PCR assays with primers designed for the genes involved in the atrazine degradation pathway. Positive results in the amplification were found in at least three of these bacteria (Arthrobacter sp., Nocardioides sp. and Pseudomonas sp.). The atrazine-degrading genetic potential was shown to be dependent on the trzN and atzA,B,C gene combination. These results suggest for the first time the adaptation of the bacterial population present in deep aquifer zones to atrazine exposure, even after more than 15 years of its ban in Spain. In addition, this study provides the baseline data about the bacterial communities found in deep aquifer zones from the hypersaline lake-aquifer system.


2004 ◽  
Vol 54 (3) ◽  
pp. 935-940 ◽  
Author(s):  
Stuart P. Donachie ◽  
John P. Bowman ◽  
Maqsudul Alam

A Gram-negative bacterium designated LA1T was isolated from water collected in hypersaline Lake Laysan on Laysan Island in the Northwestern Hawaiian Islands. Cells occurred singly as fine rods to short filaments. Growth in 50 % strength marine broth occurred optimally when the medium contained 7·5–10 % (w/v) NaCl. The major fatty acids in LA1T grown at 15 and 30 °C were 12-methyl tetradecanoic acid and 13-methyl tetradecanoic acid, respectively. The nucleotide sequence of the 16S rRNA gene showed that LA1T belonged in the Cytophaga–Flavobacterium–Bacteroides (CFB) group in the domain Bacteria. The closest described neighbour in terms of 16S rRNA gene sequence identity was Psychroflexus torquis ACAM 623T (94·4 % over 1423 bases), an obligate psychrophile from Antarctic sea-ice. The G+C content of 35·0 mol% was consistent with this affiliation. Phenotypic and genotypic analyses, including DNA hybridization, indicated that LA1T could be assigned to the genus Psychroflexus but, based on significant differences, including growth at 43 °C, it constitutes a novel species, Psychroflexus tropicus sp. nov., for which LA1T (=ATCC BAA-734T=DSM 15496T) is the type strain.


2013 ◽  
Vol 63 (Pt_1) ◽  
pp. 345-351 ◽  
Author(s):  
Mohammad Ali Amoozegar ◽  
Maryam Bagheri ◽  
Maryam Didari ◽  
Seyed Abolhassan Shahzedeh Fazeli ◽  
Peter Schumann ◽  
...  

A novel Gram-positive, moderately halophilic bacterium, designated strain X4BT, was isolated from soil around the hypersaline lake Aran-Bidgol in Iran and characterized taxonomically using a polyphasic approach. Cells of strain X4BT were motile rods and formed ellipsoidal endospores at a terminal or subterminal position in swollen sporangia. Strain X4BT was a strictly aerobic bacterium, catalase- and oxidase-positive. The strain was able to grow at NaCl concentrations of 0.5–22.5 % (w/v), with optimum growth occurring at 7.5 % (w/v) NaCl. The optimum temperature and pH for growth were 35 °C and pH 7.0. Analysis of 16S rRNA gene sequence revealed that strain X4BT is a member of the family Bacillaceae , constituting a novel phyletic lineage within this family. Highest sequence similarities were obtained with the 16S rRNA gene sequences of the type strains of Sediminibacillus albus (96.0 %), Paraliobacillus ryukyuensis (95.9 %), Paraliobacillus quinghaiensis (95.8 %) and Sediminibacillus halophilus (95.7 %), respectively. The DNA G+C content of this novel isolate was 35.2 mol%. The major cellular fatty acids of strain X4BT were anteiso-C15 : 0 and anteiso-C17 : 0 and its polar lipid pattern consisted of diphosphatidylglycerol, phosphatidylglycerol, two aminolipids, an aminophospholipid and an unknown phospholipid. The isoprenoid quinones were MK-7 (89 %) and MK-6 (11 %). The peptidoglycan contained meso-diaminopimelic acid as the diagnostic diamino acid. On the basis of 16S rRNA gene sequence analysis in combination with chemotaxonomic and phenotypic data, strain X4BT represents a novel species in a new genus in the family Bacillaceae , order Bacillales for which the name Saliterribacillus persicus gen. nov., sp. nov. is proposed. The type strain of the type species (Saliterribacillus persicus) is X4BT ( = IBRC-M 10629T = KCTC 13827T).


2006 ◽  
Vol 56 (6) ◽  
pp. 1257-1261 ◽  
Author(s):  
Liming Wang ◽  
Ying Huang ◽  
Zhiheng Liu ◽  
Michael Goodfellow ◽  
Carlos Rodríguez

The taxonomic position of ten acidophilic actinomycetes isolated from an acidic rice-field soil was established using a polyphasic approach. 16S rRNA gene sequences determined for the isolates were aligned with corresponding sequences of representatives of the genera Kitasatospora, Streptacidiphilus and Streptomyces and phylogenetic trees were inferred using four tree-making algorithms. The isolates had identical sequences and formed a distinct branch at the periphery of the Streptacidiphilus 16S rRNA gene tree. The chemotaxonomic and morphological properties of representative isolates were consistent with their assignment to the genus Streptacidiphilus. The isolates shared nearly identical phenotypic profiles that readily distinguished them from representatives of the established species of Streptacidiphilus. It is evident from the genotypic and phenotypic data that the isolates form a homogeneous group that corresponds to a novel species in the genus Streptacidiphilus. The name proposed for this new taxon is Streptacidiphilus oryzae sp. nov.; the type strain is strain TH49T (=CGMCC 4.2012T=JCM 13271T).


2010 ◽  
Vol 60 (8) ◽  
pp. 1802-1806 ◽  
Author(s):  
M. C. Gutiérrez ◽  
A. M. Castillo ◽  
P. Corral ◽  
H. Minegishi ◽  
A. Ventosa

Two novel haloalkaliphilic archaea, strains CG-6T and CG-4, were isolated from sediment of the hypersaline Lake Chagannor in Inner Mongolia, China. Cells of the two strains were pleomorphic, non-motile and strictly aerobic. They required at least 2.5 M NaCl for growth, with optimum growth at 3.4 M NaCl. They grew at pH 8.0–11.0, with optimum growth at pH 9.0. Hypotonic treatment with less than 1.5 M NaCl caused cell lysis. The two strains had similar polar lipid compositions, possessing C20C20 and C20C25 derivatives of phosphatidylglycerol and phosphatidylglycerol phosphate methyl ester. No glycolipids were detected. Comparison of 16S rRNA gene sequences and morphological features placed them in the genus Natronorubrum. 16S rRNA gene sequence similarities to strains of recognized species of the genus Natronorubrum were 96.2–93.8 %. Detailed phenotypic characterization and DNA–DNA hybridization studies revealed that the two strains belong to a novel species in the genus Natronorubrum, for which the name Natronorubrum sediminis sp. nov. is proposed; the type strain is CG-6T (=CECT 7487T =CGMCC 1.8981T =JCM 15982T).


2014 ◽  
Vol 64 (Pt_5) ◽  
pp. 1629-1634 ◽  
Author(s):  
Mustafa Camas ◽  
Aysel Veyisoglu ◽  
Nevzat Sahin

A novel Gram-reaction-positive, rod-shaped, non-motile and mycolic acid-containing strain, CR3272T, isolated from soil, was studied using a polyphasic approach. The organism showed a combination of chemotaxonomic and morphological properties typical of the genus Nocardia . The cell wall contained meso-diaminopimelic acid (type IV) and whole-cell sugars were galactose, glucose, arabinose and xylose. The predominant menaquinone was MK-8(H4cyc). The major phospholipids were diphosphatidylglycerol and phosphatidylinositol mannosides. Major fatty acids were C16 : 0, C18 : 1 cis9, C18 : 0 10-methyl (TBSA) and C16 : 1 cis9. The novel strain formed distinct phyletic line in the Nocardia 16S rRNA gene tree and was closely associated with Nocardia goodfellowii A2012T (98.6 % 16S rRNA gene sequence similarity), Nocardia alba YIM 30243T (98.5 %) and Nocardia caishijiensis F829T (97.9 %). However, DNA–DNA relatedness values and phenotypic data demonstrated that strain CR3272T was clearly distinguished from all closely related species of the genus Nocardia . It is concluded that the organism be classified as representing a novel species of the genus Nocardia , for which the name Nocardia sungurluensis is proposed. The type strain is CR3272T ( = DSM 45714T = KCTC 29094T).


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