Tengchong Hot Springs Research Articles

Streptomyces calidiresistens sp. nov., isolated from a hot spring sediment.

A Streptomyces-like actinomycete strain, designated as YIM 78087(T), was isolated from a sediment sample collected from Hehua hot spring in Tengchong, Yunnan province, south-west China. The taxonomic position of strain YIM 78087(T) was investigated by a polyphasic approach. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain YIM 78087(T) belongs to the genus Streptomyces and is closely related to Streptomyces fimbriatus DSM 40942(T), Streptomyces marinus DSM 41968(T) and Streptomyces qinglanensis DSM 42035(T) (97.18, 97.05 and 97.1% similarity, respectively). Combined with the low values of DNA-DNA hybridization between strain YIM 78087(T) and its closest neighbours, these analyses indicated that this new isolate represents a different genomic species in the genus Streptomyces. The predominant menaquinones of strain YIM 78087(T) were identified as MK-9 (H4) and MK-9 (H6). The major fatty acids were identified as anteiso-C15:0 (28.4%), anteiso-C17:0 (23.0%) and iso-C16:0 (15.1%). The whole-cell hydrolysates found to contain glucose, mannose and ribose. The DNA G+C content was determined to be 73.0mol%. Based on the comparative analysis of phenotypic and genotypic characteristics, it is proposed that strain YIM 78087(T) represents a novel species of the genus Streptomyces, for which the name Streptomyces calidiresistens sp. nov., is proposed. The type strain is YIM 78087(T) (=BCRC 16955(T)=DSM 42108(T)=JCM 19629(T)).

Optimization of titanium dioxide enrichment of phosphopeptides and application in the <I>Thermoanaerobacter tengcongensis</I> phosphoproteome analysis

Using titanium dioxide is a very good strategy for the phosphopeptide enrichment. There are many other factors can affect the enrichment efficiency, and the optimization of parameters was needed for better enrichment results. In this study, the peptide mixtures of six standard proteins were used as the model samples to evaluate and optimize the parameters such as the proportion of acetonitrile and trifluoroacetic acid in loading buffer and the TiO2-to-peptide ratio. The results showed that 80% (v/v) acetonitrile, 1% (v/v) trifluoroacetic acid and 40 : 1 (m/m) TiO2-to-peptide ratio were the optimum parameters to obtain the best enrichment selectivity and maximum phosphopeptides identification. For the first time, the optimum enrichment conditions were applied for the phosphoproteome analysis of the Thermoanaerobacter tengcongensis, an anaerobic, saccharolytic, thermophilic bacterium isolated from a hot spring in Tengchong, China, and 25 phosphorylated proteins were identified in the preliminary experiment. The results provided a reference for further study on this organism survived under extreme environment.

A Comprehensive Census of Microbial Diversity in Hot Springs of Tengchong, Yunnan Province China Using 16S rRNA Gene Pyrosequencing

The Rehai and Ruidian geothermal fields, located in Tengchong County, Yunnan Province, China, host a variety of geochemically distinct hot springs. In this study, we report a comprehensive, cultivation-independent census of microbial communities in 37 samples collected from these geothermal fields, encompassing sites ranging in temperature from 55.1 to 93.6°C, in pH from 2.5 to 9.4, and in mineralogy from silicates in Rehai to carbonates in Ruidian. Richness was low in all samples, with 21–123 species-level OTUs detected. The bacterial phylum Aquificae or archaeal phylum Crenarchaeota were dominant in Rehai samples, yet the dominant taxa within those phyla depended on temperature, pH, and geochemistry. Rehai springs with low pH (2.5–2.6), high temperature (85.1–89.1°C), and high sulfur contents favored the crenarchaeal order Sulfolobales, whereas those with low pH (2.6–4.8) and cooler temperature (55.1–64.5°C) favored the Aquificae genus Hydrogenobaculum. Rehai springs with neutral-alkaline pH (7.2–9.4) and high temperature (>80°C) with high concentrations of silica and salt ions (Na, K, and Cl) favored the Aquificae genus Hydrogenobacter and crenarchaeal orders Desulfurococcales and Thermoproteales. Desulfurococcales and Thermoproteales became predominant in springs with pH much higher than the optimum and even the maximum pH known for these orders. Ruidian water samples harbored a single Aquificae genus Hydrogenobacter, whereas microbial communities in Ruidian sediment samples were more diverse at the phylum level and distinctly different from those in Rehai and Ruidian water samples, with a higher abundance of uncultivated lineages, close relatives of the ammonia-oxidizing archaeon “Candidatus Nitrosocaldus yellowstonii”, and candidate division O1aA90 and OP1. These differences between Ruidian sediments and Rehai samples were likely caused by temperature, pH, and sediment mineralogy. The results of this study significantly expand the current understanding of the microbiology in Tengchong hot springs and provide a basis for comparison with other geothermal systems around the world.

Complete Genome Sequence of Alicyclobacillus acidocaldarius Strain Tc-4-1

Alicyclobacillus acidocaldarius strain Tc-4-1 was initially isolated from a hot spring in Tengchong, China. This organism is both thermophilic and acidophilic. It can produce heat- and acid-stable enzymes, such as amylase and esterase, which may be important in industry. Here we report the whole genome sequence of the strain.

Biological Dark Matter Exerts Irresistible Pull in Yunnan

Researchers hope a 5-year study launched this summer at the Rehai hot springs in Tengchong, a volcanic landscape in southwest China's Yunnan Province, will yield clues about the hundreds of unknown organisms they harbor.

Metallosphaera cuprina sp. nov., an acidothermophilic, metal-mobilizing archaeon

A novel acidothermophilic archaeon, strain Ar-4(T), was isolated from a sulfuric hot spring in Tengchong, Yunnan, China. Cells of strain Ar-4(T) were Gram-staining-negative, irregular cocci and motile by means of flagella. Strain Ar-4(T) grew over a temperature range of 55-75 °C (optimum, 65 °C), a pH range of 2.5-5.5 (optimum, pH 3.5) and a NaCl concentration range of 0-1 % (w/v). The novel strain was aerobic and facultatively chemolithoautotrophic. The strain could extract metal ions from sulfidic ore. It was also able to oxidize reduced sulfur compounds. In addition, it was able to use heterogeneous organic materials for organotrophic growth. The main cellular lipids were calditoglycerocaldarchaeol (CGTE) and caldarchaeol (DGTE). The DNA G+C content of the strain was 40.2 mol%. Analysis of 16S rRNA gene sequences showed that strain Ar-4(T) was phylogenetically related to members of the genus Metallosphaera and had sequence similarities of 97.7 %, 97.0 % and 96.8 % with Metallosphaera hakonensis DSM 7519(T), Metallosphaera sedula DSM 5348(T) and Metallosphaera prunae DSM 10039(T), respectively. Strain Ar-4(T) showed DNA-DNA relatedness values of 47.5 %, 30.8 % and 29.1 % with M. hakonensis DSM 7519(T), M. sedula DSM 5348(T) and M. prunae DSM 10039(T), respectively. The differences in cell motility, the temperature and pH ranges for growth, the ability to utilize carbon sources, the DNA G+C content, and the low DNA-DNA relatedness values distinguished strain Ar-4(T) from recognized species of the genus Metallosphaera. On the basis of these results, it was concluded that strain Ar-4(T) represents a novel species of the genus Metallosphaera, for which the name Metallosphaera cuprina is proposed. The type strain is Ar-4(T) ( = JCM 15769(T) = CGMCC 1.7082(T)).

Isolation and characterization of an extremely long tail Thermus bacteriophage from Tengchong hot springs in China

Thermus strains are regarded as models to investigate the mechanism of thermostability of thermophiles, and phages from Thermus are particularly interesting because of their way to regulate gene expression. In this research, a Thermus bacteriophage named TSP4 (Thermus Siphoviridae phage) was isolated from Tengchong hot springs in China, and characteristics of morphology, temperature for phage production, pH and organic solvent sensitivity, DNA restriction endonuclease digestion and protein composition of TSP4 were further studied. TSP4 belonged to the Siphoviridae family and had a hexagonal head of 73 nm in diameter, an extremely long and flexible tail of 785 nm in length and 10 nm in width. TSP4 was very stable at 65 °C and pH 7.6. The capsid was apparently devoid of lipid. By SDS-PAGE, six protein bands were found in purified virions. Despite their exceptional habitats separated by thousands of kilometers, the characteristics of this thermophilic phage showed high similarity to Thermus siphoviruses P23-45 and P74-26 isolated from Kamchatka peninsula hot springs in the Far East, Russia.

Diversity of Crenarchaeota in terrestrial hot springs in Tengchong, China

Diversity of Crenarchaeota was investigated in eight terrestrial hot springs (pH 2.8-7.7; temperature 44-96 degrees C) located in Tengchong, China, using 16S rRNA gene phylogenetic analysis. A total of 826 crenarchaeotal clones were sequenced and a total of 47 operational taxonomic units (OTUs) were identified. Most (93%) of the identified OTUs were closely related (89-99%) to those retrieved from hot springs and other thermal environments. Our data showed that temperature may predominate over pH in affecting crenarchaeotal diversity in Tengchong hot springs. Crenarchaeotal diversity in moderate-temperature (59-77 degrees C) hot springs was the highest, indicating that the moderately hot-temperature springs may provide optimal conditions for speciation of Crenarchaeota.

Anoxybacillus tengchongensis sp. nov. and Anoxybacillus eryuanensis sp. nov., facultatively anaerobic, alkalitolerant bacteria from hot springs

Two novel thermophilic, spore-forming bacterial strains, T-11(T) and E-112(T), were isolated from hot springs in Tengchong and Eryuan counties of Yunnan province in south-west China. The strains were Gram-stain-positive rods, occurring singly or in chains. Growth of strain T-11(T) was observed between 30 and 75 °C (optimum 50 °C) and at pH 7-11 (optimum pH 8.5), while the temperature range for strain E-112(T) was 35-70 °C (optimum 55 °C) and the pH range was 7.0-11.0 (optimum pH 8.0). The DNA G+C contents of strains T-11(T) and E-112(T) were 41.1 and 42.6 mol%, respectively. On the basis of 16S rRNA gene sequence similarity, the two strains were shown to be related most closely to Anoxybacillus species. The chemotaxonomic characteristics [predominant isoprenoid quinone menaquinone 7 (MK-7); major fatty acids iso-C(15 : 0) and iso-C(17 : 0)] also supported the affiliation of strains T-11(T) and E-112(T) to the genus Anoxybacillus. The results of DNA-DNA hybridization and physiological and biochemical tests allowed genotypic and phenotypic differentiation of strains T-11(T) and E-112(T) from Anoxybacillus species with validly published names. Strains T-11(T) and E-112(T) therefore represent two novel species, for which the names Anoxybacillus tengchongensis sp. nov. (type strain T-11(T) =CCTCC AB209237(T) =KCTC 13721(T)) and Anoxybacillus eryuanensis sp. nov. (type strain E-112(T) =CCTCC AB209236(T) =KCTC 13720(T)) are proposed.

Actinobacterial Diversity in Hot Springs in Tengchong (China), Kamchatka (Russia), and Nevada (USA)

Actinobacterial diversity was investigated in three hot springs [Tengchong Frog Mouth (TFM) hot spring (in Tengchong, China), Kamchatka Robb Flag hot spring (in Kamchatka, Russia), and Nevada Boiling Spring (in Nevada, USA)], a Tengchong Frog Mouth hot spring-derived pond and a soil sample near the TFM hot spring. Culture-independent techniques were employed including denaturing gradient gel electrophoresis and actinobacterial 16S rRNA gene phylogenetic analysis. The three hot springs and the hot spring-derived pond had pH of 6.8 to 7.9 and temperature of 50 to 81°C. Actinobacteria were for the first time found to be very diverse in hot springs with temperature up to 81°C, indicating that they have an unexpected capability of adapting to hot spring environments. A total of 346 actinobacterial clones were obtained from the three hot springs, and these clones were identified as 31 RFLP types, falling into the Actinomycetales (only limit to the Frankineae suborder), the Rubrobacterales, uncultured Candidatus Microthrix, unclassified Actinobacteria, and uncultured Candidate Division OP10. Our data have implications for a better understanding of the distribution of Actinobacteria in hot springs.

Growth and surface properties of new thermoacidophilic Archaea strain Acidianus manzaensis YN-25 grown on different substrates

The growth and surface properties of new thermoacidophilic Archaea strain Acidianus manzaensis YN-25 isolated from an acid hot spring in Tengchong, Yunnan Province, China were investigated cultured on different substrates including soluble substrate ferrous sulfate and nonsoluble solid substrates S 0, pyrite and chalcopyrite. The growth characteristics of the cells in each substrate were characterized with the changes in cell number, pH, E h, and concentrations of Fe 2+ or SO 4 2−, or ratios of [Fe 2+] to [Fe 3+], and the surface properties were characterized and analyzed in terms of Zeta-potential, hydrophobicity, and surface FT-IR spectra of the cells. The results show that the cells grown on solid substrates have higher value of isoelectric points. They are more hydrophobic and express more surface proteins than ferrous sulfate grown cells.

Isolation and characterization of YNTC-1, a novel Alicyclobacillus sendaiensis strain

A heterotrophic acidothermophilic bacterial strain, YNTC-1, was isolated from an acidic hot spring in Tengchong, Yunan, China. YNTC-1 grows at pH value of 1.5–8.0 and temperature of 40–70 °C, with optimal pH and temperature at 3.0 and 55 °C, respectively. The cells of the strain are in shape of short rod, with 1.0–1.2 μm in length and 0.7–0.8 μm in diameter, and with distinct spores at both poles of each cell. The predominant fatty acids in cellular membrane of the strain are C18:1 θ7c. 16s rRNA gene analysis reveals that this strain is closely related to Alicyclobacillus sendaiensis, with over 99% sequence similarity. Based on phenotypic and genotypic analyses, YNTC-1 is identified as a member of A. sendaiensis. Considering some important morphological and biochemical differences between strain YNTC-1 and A. sendaiensis ATCC 27009T, YNTC-1 may be proposed to be a novel subspecies of A. sendaiensis. However, this viewpoint has to be confirmed by further studies. Co-bioleaching of pyrite and chalcopyrite with strain YN22, Sulfobacillus thermosulfidooxidans, shows that strain YNTC-1 has no evident influence on bioleaching rates of these two sulphide minerals.

Sulfolobus tengchongensis sp. nov., a novel thermoacidophilic archaeon isolated from a hot spring in Tengchong, China.

A novel thermoacidophilic strain, designated RT8-4, was isolated from an acidic hot spring in Tengchong, Yunnan, China, and characterized phenotypically and phylogenetically. Cells of strain RT8-4 are irregular cocci with peritrechous flagella. The strain grows aerobically in either a lithotrophic or a heterotrophic mode. No anaerobic growth is apparent. Growth on elemental sulfur occurs through the oxidation of sulfur. Strain RT8-4 is capable of utilizing tryptone, d-xylose, d-arabinose, d-galactose, maltose, sucrose, d-fructose, or l-glutamic acid as the sole source of carbon. d-Glucose and d-mannose are not utilized. RT8-4 grows optimally at 85 degrees C and pH 3.5. The G+C content of the genome of RT8-4 is 34.4 mol%. Phylogenetic analysis based on 16S rDNA sequence as well as DNA-DNA hybridization and phenotypic characterization identifies strain RT8-4 as a novel species in the genus Sulfolobus. It is proposed that strain RT8-4 be designated as Sulfolobus tengchongensis sp. nov. The type strain is RT8-4(T).

Meiothermus rosaceus sp. nov. isolated from Tengchong hot spring in Yunnan, China

A rosy-pigmented Gram-negative, thermophilic bacterium with an optimum growth temperature of about 55°C was isolated from Tengchong hot springs in Yunnan province, China. Its growth scarcely occurred below 40°C or above 70°C. Phylogenetic and secondary structural analyses of 16S rRNA and DNA–DNA hybridization showed that the organism represented a new species of the genus Meiothermus. This new species could be distinguished easily from other species of the genus Meiothermus by the following phenotypic characteristics: rosy pigment, expanded body, sucrose and maltose were not utilized, gelatin and starch were not hydrolyzed. On the basis of the above data, the name Meiothermus rosaceus sp. nov. was proposed for the species represented by the strain RH9901 T(CCTCC-AB200291).

Meiothermus rosaceus sp nov isolated from Tengchong hot spring in Yunnan, China.

A rosy-pigmented Gram-negative, thermophilic bacterium with an optimum growth temperature of about 55 degrees C was isolated from Tengchong hot springs in Yunnan province, China. Its growth scarcely occurred below 40 degrees C or above 70 degrees C. Phylogenetic and secondary structural analyses of 16S rRNA and DNA-DNA hybridization showed that the organism represented a new species of the genus Meiothermus. This new species could be distinguished easily from other species of the genus Meiothermus by the following phenotypic characteristics: rosy pigment, expanded body, sucrose and maltose were not utilized, gelatin and starch were not hydrolyzed. On the basis of the above data, the name Meiothermus rosaceus sp. nov. was proposed for the species represented by the strain RH9901(T)(CCTCC-AB200291).

A complete sequence of the T. tengcongensis genome.

Thermoanaerobacter tengcongensis is a rod-shaped, gram-negative, anaerobic eubacterium that was isolated from a freshwater hot spring in Tengchong, China. Using a whole-genome-shotgun method, we sequenced its 2,689,445-bp genome from an isolate, MB4(T) (Genbank accession no. AE008691). The genome encodes 2588 predicted coding sequences (CDS). Among them, 1764 (68.2%) are classified according to homology to other documented proteins, and the rest, 824 CDS (31.8%), are functionally unknown. One of the interesting features of the T. tengcongensis genome is that 86.7% of its genes are encoded on the leading strand of DNA replication. Based on protein sequence similarity, the T. tengcongensis genome is most similar to that of Bacillus halodurans, a mesophilic eubacterium, among all fully sequenced prokaryotic genomes up to date. Computational analysis on genes involved in basic metabolic pathways supports the experimental discovery that T. tengcongensis metabolizes sugars as principal energy and carbon source and utilizes thiosulfate and element sulfur, but not sulfate, as electron acceptors. T. tengcongensis, as a gram-negative rod by empirical definitions (such as staining), shares many genes that are characteristics of gram-positive bacteria whereas it is missing molecular components unique to gram-negative bacteria. A strong correlation between the G + C content of tDNA and rDNA genes and the optimal growth temperature is found among the sequenced thermophiles. It is concluded that thermophiles are a biologically and phylogenetically divergent group of prokaryotes that have converged to sustain extreme environmental conditions over evolutionary timescale.