Research On The Characteristics Of Verrucomicrobiota Based On Genomic Analysis And Isolation And Cluture Of Marine Verrucomicrobiota

The distribution of Verrucomicrobiota is very common,widely present in marine,terrestrial and host environments,participating in the process of element circulation,organic degradation and nutrient cycling,and playing an important role in biogeochemical cycles,but there are fewer cultureable strains of Verrucomicrobiota,and there are a large number of uncultured taxa in nature,which need to be explored.In this paper,the global distribution,habitat preference and genome of Verrucomicrobiota were analyzed and explored,and the following results were achieved:1.Through 10,000 sample data from the Earth microbiome project to explore the global distribution of Verrucomicrobiota and various groups of environmental bias,the results show that the distribution of Verrucomicrobiota is very wide,exist in the global 17 environmental types.The number of bacteria and the relative abundance of OTUs in the environment were 2.99%and 3.73%,respectively,ranking the seventh and sixth in bacterial phylum,which were the main groups in the environment.Through statistics and comparison of Verrucomicrobiota at the genus level,it is found that different genera have obvious habitat bias.Genus opitutus has the highest relative abundance in the rhizosphere of plants,followed by distribution in the soil.Genus coraliomargarita prefers to live in salt environment.Genus pelagicoccus has the highest abundance in salt environment,genus puniceicoccus only exists in high salt environment and salty water bodies,genus terrimicrobium is more inclined to exist in soil,and genus limisphaera has the highest abundance in fresh water.Genus akkermansia is present in the intestines and feces of animals,suggesting that a significant proportion of the bacteria within the Verrucomicrobiota have environmental preferences.2.Through the construction of phylogenetic tree of Verrucomicrobiota genome and metagenomic assembly genome,it is found that the size and GC content of genome are related to the living environment and the distance of genetic relationship.There are a large number of uncultured Verrucomicrobiota groups in nature,and there are a large number of microbial resources available for mining.3.Through the annotation of the metabolic pathway of Verrucomicrobiota genome,it was found that Verrucomicrobiota has a variety of metabolic capabilities,including glycolysis,TCA cycle,pentose phosphate pathway and glyoxylic acid cycle carbon metabolic pathway,in addition to methane oxidation,nitrogen fixation,nitrification,assimilation of sulfate reduction,assimilation and nitrate reduction.They play an important role in participating in the geochemical cycle.4.Through the annotation of the gene function of the Verrucomicrobiota genome,it was found that the Verrucomicrobiota group has the ability to synthesize a variety of secondary metabolites,including terpene,type Ⅰ polyketo synthase,type Ⅲ polyketo synthase,aryl polyene,non-ribosome polypeptide synthase,non-ribosome polypeptide synthase analogues,etc.Verrucomicrobiota encodes a variety of peptidases,which may be involved in the decomposition of peptides,including the C01B,C11A,M03A,M13,M20,M61,S01C,S15,S16,S41A,S46 family,etc.In different environments,Verrucomicrobiota have different salt tolerance mechanisms to balance their cytoplasm with the osmotic pressure in the environment.Through the Verrucomicrobiota genome polysaccharide use gene and gene cluster annotation,and the polysaccharide degradation experiment of 19 Verrucomicrobiota was verified,found that many bacteria in the Verrucomicrobiota polysaccharide degradation gene and gene cluster,and has the ability to degrade sulfated polysaccharide,starch,cellulose and sodium alginate.5.By analyzing the 16S rRNA gene amplification results of 816 samples(including seawater,kelp,coastal sediment,salt farm and salt lake tested in the laboratory,the samples of Kelp,Wendeng salt farm and Xiaoshi Island were selected for subsequent bacterial separation experiments.Finally,7 different species of verruca microbacteria(including 5 potential new strains)were isolated,and they distributed in two classes and three genera.The three potential isolates were classified into two new species of the genus Pelagicoccus,namely,SDUM812002T and SDUM812005T.Strain SDUM461003T are one new species of the genus Coraliomargarita,and their morphologies were observed by scanning electron microscopy and transmission electron microscopy.