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Natrialba hulunbeirensis sp. nov. and Natrialba chahannaoensis sp. nov., novel haloalkaliphilic archaea from soda lakes in Inner Mongolia Autonomous Region, China.

Two haloalkaliphilic archaeal strains, X21T and C112T, were isolated from soda lakes in Inner Mongolia Autonomous Region, China. Their morphology, physiology, biochemical features, polar lipid composition and 16S rRNA genes were characterized in order to elucidate their taxonomy. According to these data, strains X21T and C112T belong to the genus Natrialba, although there are clear differences with respect to their physiology and polar lipid composition between the two strains and the type species, Natrialba asiatica. On the basis of low DNA-DNA hybridizations, these two strains should be considered as new species of genus Natrialba. The names Natrialba hulunbeirensis sp. nov. (type strain X21T = AS 1.1986T = JCM 10989T) and Natrialba chahannaoensis sp. nov. (type strain C112T = AS 1.1977T = JCM 10990T) are proposed.

Base Composition↗

Halorubrum tebenquichense sp. nov., a novel halophilic archaeon isolated from the Atacama Saltern, Chile.

A novel extremely halophilic archaeon was isolated from Lake Tebenquiche, situated in the northern part of the Atacama Saltern, Chile. The cells of these micro-organisms were mostly irregularly disc-shaped. They grew in medium containing saturated concentrations of NaCl and did not require magnesium for optimal growth. The polar lipid composition revealed the presence of mannosyl-2-sulfate-(1-4)-glycosyl-archaeol, the main glycolipid of the genus Halorubrum, and two new glycolipids. The G+C content of the DNA was 63.2 mol%. Phylogenetic analysis of the 16S rRNA gene placed strain ALT6-92T within the Halorubrum cluster. The low DNA-DNA hybridization value justified classification in a new species for which the name Halorubrum tebenquichense sp. nov. is proposed. The type strain is ALT6-92T (= CECT 5317T = DSM 14210T).

Base Composition↗

Association of newly discovered IS elements with the dichloromethane utilization genes of methylotrophic bacteria.

Dichloromethane (DCM) dehalogenases enable facultative methylotrophic bacteria to utilize DCM as sole carbon and energy source. DCM-degrading aerobic methylotrophic bacteria expressing a type A DCM dehalogenase were previously shown to share a conserved 4.2 kb BamHI DNA fragment containing the dehalogenase structural gene, dcmA, and dcmR, the gene encoding a putative regulatory protein. Sequence analysis of a 10 kb DNA fragment including this region led to the identification of three types of insertion sequences identified as IS1354, IS1355 and IS1357, and also two ORFs, orf353 and orf192, of unknown function. Two identical copies of element IS1354 flank the conserved 4.2 kb fragment as a direct repeat. The occurrence of these newly identified IS elements was shown to be limited to DCM-utilizing methylotrophs containing a type A DCM dehalogenase. The organization of the corresponding dcm regions in 12 DCM-utilizing strains was examined by hybridization analysis using IS-specific probes. Six different groups could be defined on the basis of the occurrence, position and copy number of IS sequences. All groups shared a conserved 5.6 kb core region with dcmA, dcmR, orf353 and orf192 as well as IS1357. One group of strains including Pseudomonas sp. DM1 contained two copies of this conserved core region. The high degree of sequence conservation observed within the genomic region responsible for DCM utilization and the occurrence of clusters of insertion sequences in the vicinity of the dcm genes suggest that a transposon is involved in the horizontal transfer of the DCM-utilization character among methylotrophic bacteria.

Amino Acid Sequence↗

Influence of salt on the transcription of the gas-vesicle genes of Haloferax mediterranei and identification of the endogenous transcriptional activator gene.

The transcription of the 14 gvp genes of the gas-vesicle-encoding mc-vac region was investigated, using RNA from 25% and 15% (w/v) salt cultures of the moderately halophilic archaeon Haloferax mediterranei. Transcription occurred only from two promoters, located in front of the mc-gvpA and mc-gvpD genes. In both cultures transcripts spanning the entire mc-gvpDEFGHIJKLM transcription unit were formed only during the exponential growth phase. Amounts of these transcripts were larger in the 25% salt culture, in which the 2.0 kb mc-gvpD transcripts were also synthesized during the stationary phase. The levels of the mc-gvpD transcripts and of the 324 nt mc-gvpA mRNA increased in parallel during the stationary phase of the 25% salt culture. Only under these conditions were mRNAs spanning the entire mc-gvpACNO transcription unit observed, and gas-vesicles were formed. Investigation of the influence of the mc-gvpDE genes on both mc-vac promoters in transformants revealed that by themselves they were nearly inactive. The addition of mc-gvpE, however, resulted in a high level of constitutively produced mc-gvpA and mc-gvpD mRNA, indicating a transcriptional activator function for the mc-gvpE product.

Archaeal Proteins↗

Taxonomic problems arising in the genera Haloterrigena and Natrinema.

Data currently available on members of the genera Haloterrigena and Natrinema suggest that some strains of the species Haloterrigena turkmenica that have been shown by DNA-DNA hybridization to belong to this species show a high degree of 16S rDNA sequence similarity to members of the genus NATRINEMA: However, closer examination of the data presented in the original publications and in subsequent publications suggests that there may be a number of problems associated with some of the published data.

DNA, Archaeal↗

Taxonomic study of Halorubrum distributum and proposal of Halorubrum terrestre sp. nov.

Halorubrum distributum (basonym, Halobacterium distributum) is an extremely halophilic, aerobic archaeon isolated from saline soils, which was described on the basis of phenotypic features of several strains. The designated type strain of the species (1m(T)=VKM B-1733(T)=JCM 9100(T)) was shown recently to differ from the other strains. In this study, Halorubrum distributum isolates have been characterized with regard to phenotypic features, polar lipid content, comparison of 16S rRNA gene sequences and DNA-DNA hybridization. On the basis of these data, a novel species that includes the other isolates is proposed, with the name Halorubrum terrestre sp. nov. The type strain of this novel species is 4p(T) (=VKM B-1739(T)=JCM 10247(T)). The DNA G+C content of this novel species is 64.2-64.9 mol% (64.4 mol% for the type strain).

Base Composition↗

Halorubrum tibetense sp. nov., a novel haloalkaliphilic archaeon from Lake Zabuye in Tibet, China.

A novel haloalkaliphilic archaeon, strain 8W8T, was isolated from Lake Zabuye, on the Tibetan Plateau, China. On the basis of 16S rRNA gene sequence analysis, strain 8W8T was shown to belong to the genus Halorubrum and was related to Halorubrum vacuolatum (96.7% sequence similarity), Halorubrum saccharovorum (96.0%), Halorubrum lacusprofundi (95.4%) and Halorubrum sodomense (95.3%). The phylogenetic distance from any species within the other genera of Halobacteriales was lower than 90%. The major polar lipids of strain 8W8T were C20C20 and C20C25 derivatives of phosphatidylglycerol phosphate and phosphatidylglycerol phosphate methyl ester. The results of DNA-DNA hybridization and physiological and biochemical tests allowed genotypic and phenotypic differentiation of strain 8W8T from the eight Halorubrum species with validly published names. Therefore, strain 8W8T represents a novel species, for which the name Halorubrum tibetense sp. nov. is proposed, with the type strain 8W8T (=AS 1.3239T=JCM 11889T).

China↗

Halorubrum xinjiangense sp. nov., a novel halophile isolated from saline lakes in China.

A novel halophilic archaeon, strain BD-1T, was isolated from Xiao-Er-Kule Lake in Xinjiang, China. The taxonomy of strain BD-1T was studied by polyphasic methods. According to 16S rRNA gene sequence similarity, strain BD-1T was phylogenetically related to Halorubrum trapanicum (98.4 %), Halorubrum sodomense (98.0 %), Halorubrum distributum (97.8 %) and Halorubrum coriense (97.3 %). Strain BD-1T is able to grow at 10 degrees C and further differs physiologically from the above species in the assimilation of sugars. The G + C content of DNA is 68.0 % (Tm). The DNA-DNA relatedness values to Hrr. trapanicum and Hrr. distributum are 47 and 24 %, respectively. It is concluded that strain BD-1T represents a novel species of the genus Halorubrum, for which the name Halorubrum xinjiangense sp. nov. is proposed. The type strain is BD-1T (= AS 1.3527T = JCM 12388T).

Base Composition↗

Halorubrum alkaliphilum sp. nov., a novel haloalkaliphile isolated from a soda lake in Xinjiang, China.

A novel haloalkaliphilic archaeon, strain DZ-1T, was isolated from a soda lake in Xinjiang, China. The taxonomy of strain DZ-1T was studied by polyphasic methods. According to 16S rRNA gene sequence similarity, strain DZ-1T was phylogenetically related to Halorubrum tibetense (97.5 %), Halorubrum vacuolatum (95.7 %) and Halorubrum saccharovorum (95.9 %). Strain DZ-1T was able to grow at 20-44 degrees C and was also physiologically different from the above-mentioned species with respect to assimilation of sugars and utilization of organic acids. The DNA G+C content of strain DZ-1T was 62.1 mol% (Tm). The DNA-DNA relatedness of strain DZ-1T to H. tibetense and H. vacuolatum was 22 and 13 %, respectively. It was concluded that strain DZ-1T represents a novel species of the genus Halorubrum, for which the name Halorubrum alkaliphilum (type strain, DZ-1T=AS 1.3528T=JCM 12358T) is proposed.

Base Composition↗

Natrinema altunense sp. nov., an extremely halophilic archaeon isolated from a salt lake in Altun Mountain in Xinjiang, China.

A novel extremely halophilic strain, AJ2(T), was isolated from Ayakekum salt lake located in the Altun Mountain National Nature Reserve in Xinjiang, China. This isolate was neutrophilic, motile and grew in a wide range of MgCl(2) concentrations (0.005-1.0 M). The major polar lipids of the isolate were C(20)C(20) and C(20)C(25) derivatives of phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester and phosphatidylglycerol sulfate. A comprehensive 16S rRNA gene sequence analysis revealed that the isolate shared 96.6-97.7 % sequence identity with Natrinema species. The isolate, however, could be genetically differentiated from these species by DNA-DNA hybridization analysis and on the basis of its physiological properties. On the basis of the polyphasic evidence, strain AJ2(T) (= AS 1.3731(T) = JCM 12890(T)) represents the type strain of a novel species, for which the name Natrinema altunense sp. nov. is proposed.

China↗

Halobiforma lacisalsi sp. nov., isolated from a salt lake in China.

A Gram-negative, motile, neutrophilic and extremely halophilic strain, AJ5(T), was isolated from a salt lake in Xinjiang, China, and subjected to polyphasic taxonomic study. The major polar lipids of the isolate were C(20)C(20) and C(20)C(25) derivatives of phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester and glycolipid. The DNA G+C content was 64.9 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain AJ5(T) clustered with members of the genus Halobiforma, exhibiting high sequence similarity to the 16S rRNA gene sequences of Halobiforma nitratireducens (96.3%) and Halobiforma haloterrestris (99.0%). Comparative analysis of phenotypic characteristics and DNA-DNA hybridization between strain AJ5(T) and Halobiforma species supported the conclusion that AJ5(T) represents a novel species within this genus, for which the name Halobiforma lacisalsi sp. nov. is proposed. The type strain is AJ5(T) (=CGMCC 1.3738(T)=JCM 12983(T)).

Bacterial Typing Techniques↗

Natronorubrum aibiense sp. nov., an extremely halophilic archaeon isolated from Aibi salt lake in Xin-Jiang, China, and emended description of the genus Natronorubrum.

An extremely halophilic archaeon (strain 7-3(T)) was isolated from Aibi salt lake in Xin-Jiang, China. It formed small (<1.0 mm), red, elevated colonies on salt-milk agar medium. Strain 7-3(T) was able to grow at pH 6.5-9.5 (optimum pH 8.0) and at 20-50 degrees C (optimum 45 degrees C). Analysis of its 16S rRNA gene sequence indicated that strain 7-3(T) was phylogenetically related to members of the genus Natronorubrum, with sequence similarities of 97.0 % to Natronorubrum bangense A33(T) and 95.2 % to Natronorubrum tibetense GA33(T). The G+C content of its DNA was 61.2 mol% (T(m)). Levels of DNA-DNA relatedness to Nrr. bangense and Nrr. tibetense were 45 and 37 %, respectively. It is concluded that strain 7-3(T) (=CGMCC 1.4299(T)=JCM 13488(T)) should be classified as the type strain of a novel species of the genus Natronorubrum, for which the name Natronorubrum aibiense sp. nov. is proposed. Based on the properties of Nrr. aibiense given here and of those of Nrr. bangense and Nrr. tibetense described previously, an emended description of the genus Natronorubrum is presented.

Base Composition↗

Halorubrum ezzemoulense sp. nov., a halophilic archaeon isolated from Ezzemoul sabkha, Algeria.

A novel extremely halophilic archaeon was isolated from Ezzemoul sabkha, Algeria. The strain, designated 5.1(T), was neutrophilic, motile and Gram-negative. At least 15 % (w/v) NaCl was required for growth. The isolate grew at pH 6.5-9.0, with optimum growth at pH 7.0-7.5. Mg(2+) was required for growth. Polar lipids were C(20)C(20) derivatives of phosphatidylglycerol and phosphatidylglycerol phosphate methyl ester, and phosphatidylglycerol sulfate and sulfated diglycosyl diether. The genomic DNA G+C content of strain 5.1(T) was 61.9 mol% (T(m)). Phylogenetic analysis based on comparison of 16S rRNA gene sequences revealed that strain 5.1(T) clustered with Halorubrum species. The results of DNA-DNA hybridization and biochemical tests allowed genotypic and phenotypic differentiation of strain 5.1(T) from other Halorubrum species. The name Halorubrum ezzemoulense sp. nov. (type strain 5.1(T)=CECT 7099(T)=DSM 17463(T)) is proposed.

Algeria↗

Halorubrum lipolyticum sp. nov. and Halorubrum aidingense sp. nov., isolated from two salt lakes in Xin-Jiang, China.

Two extremely halophilic archaea, strains 9-3(T) and 31-hong(T), were isolated from Aibi salt lake and Aiding salt lake in Xin-Jiang, China. Their morphology, physiology, biochemical features, polar lipid compositions and 16S rRNA gene sequences were characterized in order to elucidate their taxonomic status. The results from this study indicated that strains 9-3(T) and 31-hong(T) are members of the genus Halorubrum. Their physiological properties and polar lipid compositions are clearly different from those of the currently described species of Halorubrum. DNA-DNA relatedness values for strain 9-3(T) with respect to its closely related neighbours Halorubrum saccharovorum JCM 8865(T) and Halorubrum lacusprofundi JCM 8891(T) were 51.6 and 25.1 %, respectively, DNA-DNA relatedness values for strain 31-hong(T) with respect to its closely related neighbours Hrr. saccharovorum JCM 8865(T) and Hrr. lacusprofundi JCM 8891(T) were 29.4 and 44.9 %, respectively, and DNA-DNA relatedness between strains 9-3(T) and 31-hong(T) was 54 %. Thus, two novel species of the genus Halorubrum are proposed, Halorubrum lipolyticum sp. nov. (type strain 9-3(T)=CGMCC 1.5332(T)=JCM 13559(T)) and Halorubrum aidingense sp. nov. (type strain 31-hong(T)=CGMCC 1.2670(T)=JCM 13560(T)).

Bacterial Typing Techniques↗

Halorubrum orientale sp. nov., a halophilic archaeon isolated from Lake Ejinor, Inner Mongolia, China.

A motile, pleomorphic, red-pigmented archaeon, strain EJ-52T, was isolated from water from Lake Ejinor, a saline lake in Inner Mongolia, China. Analysis of the almost-complete 16S rRNA gene sequence showed that the isolate was phylogenetically related to species of the genus Halorubrum, being most closely related to Halorubrum saccharovorum ATCC 29252T (96.1% sequence similarity), Halorubrum lacusprofundi JCM 8891T (95.9%), Halorubrum tibetense AS 1.3239T (95.2%), Halorubrum alcaliphilum AS 1.3528T (95.2%) and Halorubrum vacuolatum JCM 9060T (95.1%). The polar lipids of strain EJ-52T were C20C20 derivatives of phosphatidylglycerol phosphate and phosphatidylglycerol phosphate methyl ester and a sulfated diglycosyl diether. Strain EJ-52T requires at least 2.5 M NaCl for growth and grows optimally at 3.4 M NaCl. The strain grows at 25-50 degrees C, with optimal growth occurring at 35-45 degrees C. Mg2+ is not required. The DNA G+C content is 64.2 mol%. On the basis of the data obtained in this study, strain EJ52T represents a novel species, for which the name Halorubrum orientale sp. nov. is proposed. The type strain is EJ-52T (=CECT 7145T=JCM 13889T=CGMCC 1.6295T).

Base Composition↗

From genomes to function: haloarchaea as model organisms.

Haloarchaea are adapted to high-salt environments and accumulate equally high salt concentrations in the cytoplasm. The genomes of representatives of six haloarchaeal genera have been fully or partially sequenced, allowing the analysis of haloarchaeal properties in silico. Transcriptome and proteome analyses have been established for Halobacterium salinarum and Haloferax volcanii. Genetic systems are available including methods that allow the fast in-frame deletion or modification of chromosomal genes. The high-efficiency transformation system of Hf. volcanii allows the isolation of genes essential for a biological process by complementation of loss-of-function mutants. For the analysis of haloarchaeal biology many molecular genetic, biochemical, structural and cell biological methods have been adapted to application at high salt concentrations. Recently it has become clear that several different mechanisms allow the adaptation of proteins to the high salt concentration of the cytoplasm. Taken together, the wealth of techniques available make haloarchaea excellent archaeal model species.

Adaptation, Physiological↗

Living with two extremes: conclusions from the genome sequence of Natronomonas pharaonis.

Natronomonas pharaonis is an extremely haloalkaliphilic archaeon that was isolated from salt-saturated lakes of pH 11. We sequenced its 2.6-Mb GC-rich chromosome and two plasmids (131 and 23 kb). Genome analysis suggests that it is adapted to cope with severe ammonia and heavy metal deficiencies that arise at high pH values. A high degree of nutritional self-sufficiency was predicted and confirmed by growth in a minimal medium containing leucine but no other amino acids or vitamins. Genes for a complex III analog of the respiratory chain could not be identified in the N. pharaonis genome, but respiration and oxidative phosphorylation were experimentally proven. These studies identified protons as coupling ion between respiratory chain and ATP synthase, in contrast to other alkaliphiles using sodium instead. Secretome analysis predicts many extracellular proteins with alkaline-resistant lipid anchors, which are predominantly exported through the twin-arginine pathway. In addition, a variety of glycosylated cell surface proteins probably form a protective complex cell envelope. N. pharaonis is fully equipped with archaeal signal transduction and motility genes. Several receptors/transducers signaling to the flagellar motor display novel domain architectures. Clusters of signal transduction genes are rearranged in haloarchaeal genomes, whereas those involved in information processing or energy metabolism show a highly conserved gene order.

Amino Acid Motifs↗