Microbial Diversity Analysis Of Permafrost Region Dk9 Test Wells In Qianlian And Taxonmic Research For New Species Of Bacteria

Qianlian muli area is located in the northwest of Qinghai province, which is the largest coalmine region. Along with in-depth study of Muli region, with the process of gas hydrate exploration in the local area, Chinese scientists drilled out the first physical samples of natural gas hydrate. The gas source for hydrate research carried out in succession, including research of methane, butane oxidation bacteria in the surface soil. Our study is based on the longitudinal microbial community structure of the hydrate test Wells, and exploration on the key symbol microbial community of the important region. By using Ion Torrent high-throughput sequencing technologies,20 samples of bacteria and archaea 16S rRNA of vertical Wells is analyzed, and the get the following conclusions:1) According to the results of bacteria analysis, bacterial richness in Qilian mountain is pretty good, by Ion torrent high throughput sequencing,118550 sequences were gained,52805 OTUs were clustered (in total). Using two kinds of analysis method based on RDP classifier and OTUs of bacteria community structure of the diversity, similarities and differences between different samples. The figure based on RDP classifier can clearly reponsed each bacteria classification level of each sample, relative content, and can reflect the change trend of bacteria between different samples. The heatmap of constracture draw by clustering results, PCA analysis based on OTUs and Unifrac clustering obtained the similarities results. Results reveal that the bacteria in the hydrates layer and without hydrate layer get a high species richness, diversity of microbes in the extreme environment is very well. In the Hydrates layer and without hydrate layer, all contains amount of bacteria of the genus pseudomonas, but in the hydrate layer pseudomonas relative content was stable, and in the hydrate layer the relative content is not stable..In DK9-19 samples detected a large number of sphingosine, in combination with the physical properties of the samples, suggest that in the sediments which contain more heavy hydrocarbons, sphingosine growth very well. There is no obvious differences between the thydrate layer and without hydrate layer, and no obvious similarities between the hydrate layer and without hydrate layer.2) According to the analysis of archaea in qilian mountain area, archaea community structure is relatively single, archaea species richness is lower than bacteria richness. After sequencing,234928 archaea 16S rRNA sequences were obtained,35899 OTUs were clustered. Shannon index range 5.76-6.40, chao-1 index range is 1865.86 to 6249.199, but coverage of archaea is higher,0.87 to 0.97..After the analysis of the RDP classifier, the major archaea were Euryarchaeota and Crenarchaeota, in the sample of the hydrate layer and the surrounding detected methane bacteria microbes, which speculated that it can participate in methane hydrate layer flow of C metabolism. In the oil and gas showing smaple detected methane phylum micro microbes, combined with oil&gas showing effect, speculated that it may be associated with heavy hydrocarbons such as decomposition of aromatic compounds.3) After make use of different kinds analysis methods, such as genotype, biochemical characteristics, physiological, phenotype and biochemical characteristics, unclassified Z12T microorganisms categorized into genus Dyadobacter, named Dyadobacter. sediminis. Strain Z12T is gram-reaction negative, aerobic growth, no-motility, yellow colonies. After incubated on R2A plant at 25℃ for 5 days, bacteria size 0.5-0.7 x 3-5 μm. Growth well under 10-30℃, and 25℃ growth is the best; pH range is 5.5 to 10.0,7.5 to 8.0 growth is the best; the optimal growth of NaCl tolerance range is 0% to 2.0%,1.0% growth is the best. Can hydrolyze casein and twain 80, not hydrolyzed starch, tryptophan, chitin, cellulose and large molecules such as substrates, urea. Nitrate reduction is negative; C16:1ω7c and/or iso-C15:02-OH, iso-C15:o, iso-C17:1I/anteiso B is the main fatty acids; Main polar lipid is phosphatidyl ethanolamine; Major types of respiratory quinone is MK-7; Main types of pigment is yellow pigment; the genome G+C% is 45.4%.