Analysis Of Genetic Diversity Of The Methanogen During The Methane Fermentation And Isolation Of Methanogen

The methane fermentation, is a process that unites by the many kinds of microorganisms, with replacement complex biochemistry process in it. In the fermentation, The interdependence and rectriction of no-produce -methane bacterium and methanogens makes a equilibrium in the fermentation. But microorganism's metabolic activity and affection also manifests in the physical and chemical changes of material in fermentation.Methanogen is the core of the methane fermentation. And methanogen is the last member of the anaerobe chain in nature carbon cycle. The study on changes of methanogen in the methane fermentation can make us realize fermentation degree, optimize the fermentation condition and enhance the methane output. It is well known that more than 90% of the microorganisms existing in nature are refractory to selective enrichment cultures. Being anaerobic microorganism, the raise cycle of methanogen is long. These characteristics cause it extremely difficult to study community structure of methanogen through the traditional cultivation techniques.To overcome these drawbacks, molecular biological techniques especially 16SrDNA analysis are increasingly employed in the analysis of microorganisms in anaerobic natural habitats.This article has been studied the changes of chemical property of material and six major microorganisms in the methane fermentation with cow's feces as the raw material .At the same time the culture-independent 16SrDNA analysis method was used to investigate the genetic diversity of methanogen in the methane fermentation.As the same time three methanogen strains have been isolated and characterized from the fermentative fluid through Hungate techniques.1. Changes of major microorganism and chemical property of material in methane fermentationThe quantitative changes of six major microorganisms and the transformation of material's chemical property were investigated in the present study. The result indicate that the aerobic microorganisms are active and play an important role in early period of fermentation, while the anaerobic microorganisms especially methanogen are more and more remarkable later.In the fermentation the material's chemical properties such as the volatile organic acid, the entire nitrogen, PH, the redox potential have great changes. The changes are correlated with microorganisms ' changes.2. Extraction and purification of moors fluid microbial total DNA.A method of extracting moors fluid total DNA was established through the comparison of the direct and indirect extraction method. Although the extract rate of direct method is higher, the target gene is unable to be amplified by the PCR amplification from the sample even after the purification because of the high content of impurity. It is unable to carry on...