Diversity Analysis Of Spirulina And Correlative Microorganism In The Bohai Sea Coastal Wetland

Spirulina is rich in protein,carbohydrates,vitamins,unsaturated fatty acids and trace elements,has been successfully developed and applied to achieve industrial production,but with the expansion of Spirulina application,the demand for functional algal strains was also enhanced.In order to obtain the excellent algal strains,the water samples were collected for three consecutive years.The microscopic observation of the samples showed that Spirulina and Arthrospira were widely present in the Bohai Sea and its surrounding inland wetlands.But because of the effective strain concentration method has not been found,and were of single filament were often failed in pure culture,resulting in the spiral algae in some water samples of Arthrospira strains lost,unable to obtain pure culture.It is speculated that the growth of algae is affected by the physicochemical environment,so in order to obtain a pure culture of strain more,from July 2015 to September 2015,the diversity of bacteria,prokaryotic microorganisms and eukaryotic microbes were analyzed in the Bohai wetland and surrounding waters where the coastline was longer and the wetland types were diversified.One face to supplement the classification of the results of analysis,for another to provide the basis for the choice of enrichment conditions.The main results are as follows:1.The counts of Spirulina in 18 sampling sites in Bohai wetland and surrounding waters were counted,It was found that there were significant differences in the number of Spirulina in different water samples;when the water TN and TP respectively between4.9-15.8 mg/L and 0.1-0.63 mg/L,suitable for the growth of Spirulina.2.Through the Illumina HiSeq high-throughput sequencing,the dominant species of bacteria in the 18 sampling sites were Proteobacteria,Cyanobacteria,Firmicutes,and the dominant groups of eukaryotic microorganisms were Rotifera,Arthropoda,Chlorophyta,the dominant species of cyanobacteria were Synechococcus,Microcystis,Halanaerobacter,Planktothrix.Analysis of microbial diversity in 18 samples,Hebei Beidaihe wetland(B1)species richness was highest,Liaoning Panjin red Beach Bungalows and the surrounding natural wetland(L1)bacterial species diversity was the largest,Inner Mongolia Bahar Lake(N2)bacterial diversity and species richness were the lowest;Hebei Beidaihe wetland(B1)eukaryotic microbial species with highest abundance,natural wetland of Tianjin Haihe Education Park(T5)eukaryotic microbial species diversity was highest,Inner Mongolia natural wetland research town(N5)eukaryotic microbial species richness was lowest,Inner Mongolia Bayannaoer alkali Lake(N4)eukaryotic microbial species diversity was lowest;Tianjin Haihe Education Park natural ponds(T5)are the most abundant species of Cyanobacteria,the diversity of Cyanobacteria in Beidaihe wetland(B1)is higher than that in other places,and the richness and diversity of Cyanobacteria in Sobel Svatn(N3)is lower than that in other rplaces.Through the determination of the environmental factors of the 18 samples,we found that the correlation between the environmental factors and the community distribution of Cyanobacteria was from high to low: COD,TN,pH,NH3-N,chlorophyll a,DO,temperature,salinity,TP.3.In this study,the 16 S rRNA gene was amplified by using the bacteria universal primers and the Cyanobacteria specific primers from 98 strains of the laboratory.From the variation of the gene sequence and the average G+C percentage,the 16 S rRNA gene amplified by Cyanobacteria specific primers showed a large degree of genetic diversity compared with the 16 S rRNA gene amplified by bacterial universal primer.The phylogenetic tree constructed by two specific primers was divided into three groups,but in the phylogenetic development of 16 S rRNA gene amplified by bacterial universal primer,TJJ1,TJBC1,TJBC2,TJBC3,TJBC7-4,TJBC8,TJBC14-2,TJSD1 belong to the genus Arthrosophila,and 8 strains of phytoplasma belong to the genus Spirulina in the phylogenetic tree constructed by 16 S rRNA gene based on the specific primers of Cyanobacteria.Compared with the results of other indicators in this study,found the phylogenetic tree based on psbA gene sequences,strain TJBC1 and F-1070,F-351 and F-904-2 was clustered together;the phylogenetic tree based on fatty acid biomarkers characteristic index,strain TJBC3 and F-351 and F-1070 is clustered together;the results of SEM showed that the phylogenetic tree analysis results were confirmed by morphology.so the author thinks that the 16 S rRNA gene is amplified by the Cyanobacteria primers,which can be distinguished by the homology of the sequence.4.The effects of temperature,ammonia nitrogen,pH and total nitrogen were selected.In the simulation laboratory,Tianjin,Shandong,Hebei,Inner Mongolia environment,found that under the condition of experiment,the biomass of each strain was the highest,and the condition of the source was more suitable for its growth and reproduction.Therefore,in the actual production of Spirulina,combined with the natural environment and the breeding cost of the algal strains,we get the most suitable breeding conditions.