| Marine microbial resources are rich,containing a large number of species of bacteria resources,has great development and research value.However,due to the limitations of culture conditions,the resources for development and utilization are very small.In this paper,the enrichment culture method established in our laboratory was analyzed by metatranscriptomic,and concludes the classification analysis of microbial communities during enrichment culture,gene expression,metabolic pathway analysis,and functional and species annotation of metatranscriptomic.The analysis found that there were unique bacterial flora in different enrichment culture periods,indicating that the microbes responded differently to the environment in different enrichment stages.In the unenriched phase(0d),genes involved in stress and stress response are up-regulated,including the molecular chaperone DnaK,small heat shock protein(sHsp20),the molecular chaperone IbpA,the ATP-dependent Clp protease ClpB,the chaperone protein GroEL and the chaperone Protein GroES et al.In different enrichment stages,gene expression involved in citric acid cycle(TCA)was significantly increased(1.5 times and 1000 times),and the intermediate metabolite 2-oxopentate in this process played a regulatory role in the metabolism of carbon and nitrogen.In addition,genes involved in carbohydrate metabolism such as pyruvic acid and acetic acid are also up-regulated.Multiple genes involved in the aerobic respiratory pathway were highly expressed at the 0th day,and the genes of the anaerobic respiratory pathway were highly expressed during the enrichment process,indicating that aerobic respiration to anaerobic respiration occurred in the enrichment culture.In summary,most microorganisms are under stress in the environment,and enrichment culture can activate some microorganisms to increase their metabolic activity.A strain of marine Marinilabiliales was subjected to dormancy and resuscitation experiments to examine the effects of sodium pyruvate and sodiumacetate during resuscitation.Both compounds were found to have a good resuscitation effect,and the addition of sodium pyruvate and sodium acetate was the best in the recovery of the medium.A similar transformation process from low cell metabolism level to high cell metabolism level was also found in the analysis of the metatranscriptomi data of Marine sliding fungi group,which could be easily separated in the enrichment process.Combined with the preliminary experimental results,we speculated that this enrichment process could break the dormancy of Marine sliding fungi group in the original habitat.In order to further verify whether this enrichment condition is conducive to the recovery of Marine sliding fungi group,this paper induces the dormancy of Marine sliding bacteria strains screened by this method and tests the recovery effect.At the same time,it is shown that the enrichment culture method designed in this experiment has a good effect on the separation of uncultured microorganisms in the ocean.It was found that the addition of sodium pyruvate and sodium acetate was the best in the recovery of the medium.In addition,three new strains were isolated from the three sediment samples from the Weihai offshore by the enrichment culture method and several classifications were identified,which further enriched the bacterial species resources.Strain F3212~T represents a new species belonging to Oceanospirillaceae and Marinospirillum,named Marinospirillum perlucida sp.nov.,which was isolated from the a sea cucumber culture pond in weihai.It has the gene for synthesizingβ-hydroxybutyric acid(PHB),which can synthesize PHB.Strain Z0201~T represents a new species belonging to Cyclobacteriaceae and Aquiflexum,named Aquiflexum aquatilis sp.nov.,which was isolated from the lake of Zibi in Yunnan,China.Strain CQN31~T represents a new species belonging to Acetobacteraceae and Roseomonas,named Roseomonas bellus sp.nov.,which was isolated from the lake of Changqiaohai,Yunnan,China. |
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