Collection Of Bacteria Resources And Diversity Analysis Of Halophile From Wendeng Saltern And Taxonomy Analysis Of Four Novel Species

As a special habitat,salt field contains abundant microbial resources.Bioinformatics and high-throughput sequencing technology facilitate the study of microbial community structure diversity in saline environment.In this study,we use the Wendeng saltern Multi-gradient solar salt field as the research model,through the cultivation bacteria technology means and macro genome sequencing results to explore more microbial community structure of salinity gradient,the differences and the influence of environmental factors on the microbial community structure.A number of taxonomic tests were performed on the new strains isolated from the experiment.30 samples of water samples were collected from the weihai wendeng salt field(122°0'38.85" E,36°57'56.49" N),and 30 samples of mud samples were collected.Ten samples of water samples and 10 samples of mud samples were synthesized.After coarse screen and weight,the representative strain was selected for further separation and purification.We used the modified 2216E culture medium with the addition of 8%coarse salt to be cultured in the constant temperature incubator of 37?and 45 ? respectively.In the end,there were 348 strains of cultured strains,of which 79 strains of Proteobacteria were found in the sediment samples,which accounted for 47.9%.There were 11 strains of Bacteriodetes,accounting for 6.67%.There were 5 strains of Actinobacteria,accounting for 3.03%.There were 70 strains of Firmicutes,accounting for 42.4%.There were 113 strains of medium salinity,accounting for 68.5%.There were 52 strains of high salinity,accounting for 31.5%.In the sample of water samples,the strain of Proteobacteria was 33,which accounted for 70.2%.There were 5 strains of Bacteriodetes,accounting for 10.6%.There were 3 strains of Actinobacteria,which accounted for 6.4%;there were 6 strains of Firmicutes,accounting for 12.8%.There were 32 strains of medium salinity,accounting for 68.1%.There were 15 strains of high salinity,accounting for 31.9%.There were 41 potential new bacteria in the sediment sample and 23 potential new bacteria in the water sample.High-throughput sequencing technology used 519F and 915R as primers for the exploration of the community of archaea,and the archaea in water samples and sediments were amplified.Using this pair of primers,we conclude that the archaea occupies a large proportion of all the measured sequences.The percentage of archaea detected in high salinity samples was very high.In the end,there were 512 genera,289 families,185 orders,123 classes and 61 phylums.Among them,17 are the most abundant phylum,accounting for 0.03%of all sequences and 95.1%of the community.341F and 806R were used as primers to explore bacterial microbial communities,and the microbial community abundance was also analyzed for sediment samples and water samples.By using this pair of primers,the sequences in all the libraries were collated with 851 genera,466 families,270 orders,170 classes and 72 phylums.Of the 23 most abundant phylums,their sequences account for 0.03%of all sequences,but constitute 90.8%of the community,becoming the main category.CCA analysis shows the salinity is one of the most important environmental factor can adjust different salinity gradient in the salt pond prokaryotes community structure change,this experiment different salinity gradient of salt pond,salinity gradient variation range of 30 to 370 g/L.In addition to the salinity,the pH and dissolved oxygen concentration also had a significant correlation with the prokaryotic community structure.In the selection of potential new species,we selected 4 strains for further taxonomic studies.A novel heterotrophic,Gram-stain-negative bacterium,designated strain F2T,was isolated from the marine sediment collected from Weihai coastal,Shandong Province,PR China.Q-8 was the sole respiratory quinone.The major cellular fatty acid in strain F2T was iso-C15:0.The genomic DNA G+C content of the strain was 48.7 mol%.Based on the results of our polyphasic analysis,we conclude that strain F2T represents a novel species of the genus Marinicella,for which the name Marinicella sediminis sp.nov.is proposed.The type strain of the new species is F2T(=KCTC 42953T= MCCC 1H00149T).Two novel Gram-stain-negative aerobic strains,designated WDS1C4T and WDS4C29T,were isolated from marine solar saltern in Weihai,Shandong Province,PR China.Q-10 was the sole respiratory quinine of the two strains.The major cellular fatty acid in strains WDS1C4T and WDS4C29T was C18:1 ?7c.The genomic DNA G+C contents of strains WDS1C4T and WDS4C29T were 67.2 and 62.8 mol%,respectively.The similarity between WDS1C4T and WDS4C29T was 97.4%.Differential phenotypic and genotypic characteristics of the two isolates from recognized genera showed that the two strains should be classified as representing a new genus and two novel species for which the names Salibaculum Halophilum gen.nov.,sp.nov.(type strain WDS1C4T=MCCC 1H00179T =KCTC 52542T)and Salibaculum griseoflavus sp.nov.(type strain WDS4C29T =MCCC 1H00175T=KCTC 52541T)are proposed.A novel Gram-stain-negative aerobic strain,designated WFHF3C12T,was isolated from saltern of Weifang,China.Q8 was the sole respiratory quinone.The major cellular fatty acid(>10.0%)in strain WFHF3C12T was C18:1 ?7c.The genomic DNA G+C content of the strain was 66.2 mol%.Based on data from the current polyphasic study,the isolate is proposed to represent a novel species of a new genus within the family Ectothiorhodospiraceae,with the name Bradysalinae rhodinum gen.nov.sp.nov..The type strain of the type species is WFHF3C12T(=KCTC 52546T=MCCC 1H00180T).