牛粪发酵沼液中产甲烷菌的分离及分子鉴定

Molecular Identification of Methanogens Isolated from Biogas Slurry Fermented with Cow Dung

为了确定牛粪发酵沼液中疑似菌株的生物特性和种属关系。采用Hungate滚管技术、厌氧培养、革兰氏染色镜检、PCR扩增技术和抗生素耐受性鉴定等方法进行分析。实验结果表明,牛粪发酵沼液分离到了一株革兰氏阳性菌,能够利用CO_2,最适生长温度为37℃,单独或成对存在。菌落呈灰白色,光滑,表面凸起。PCR扩增出1 200 bp的16S rDNA片段,测序结果表明,该分离菌为产甲烷短杆菌。

Biological characteristics and relationship between species of Methanogens in biogas slurry fermented with cow dung were researched by Hungate rolling-tube technique,anaerobic culture method, Gram＇s method,PCR and Antibiotic resistance identification. Results showed that a gram-positive isolated bacteria was able to take advantage of the CO2,growth temperature was 37 ℃ ,existed alone or in pairs. Colonis were pale,smooth and convex. PCR results showed that 16SrDNA fragments of 1 200 bp were amplified,isolated bacteria was identified as Methanogens by DNA sequencing technology.