PCR-DGGE技术在水牛瘤胃产甲烷菌多样性分析中的应用

【目的】分析水牛瘤胃产甲烷菌的多样性,为更好地了解水牛瘤胃产甲烷菌提供科学依据。【方法】采集16头广西本地沼泽型水牛瘤胃内容物,提取总DNA,利用产甲烷菌的通用引物扩增16SrRNA基因序列进行DGGE分析,并对DGGE凝胶上7条优势条带进行回收克隆测序。【结果】共获得9条序列（5条为古菌序列、4条为非古菌序列）,5条古菌序列中有3条与甲烷短杆菌（Methanobrevibacter）的同源性高达99%、1条与甲烷短杆菌的同源性达98%、1条与广古菌门（Euryarchaeota）热原体目（Thermoplasmatales）的同源性达98%;而4条非古菌序列分别属于拟杆菌（Bac-teroidetes）、厚壁菌（Firmicutes）、变形菌门（Proteobacteria）,且彼此间同源性较小。【结论】通过PCR-DGGE技术可以较好地分析水牛瘤胃产甲烷菌的多样性。

Diversity analysis of rumen methanogens in buffalos using PCR-DGGE technique

【Objective】The present experiment was conducted to analyze the diversity of rumen methanogens in buffalos. 【Method】The rumen contents were collected from sixteen local swamp buffalo in Guangxi to extract the total DNA, and the 16S rRNA gene sequences were amplified and analyzed using universal primers and DGGE technique. The obtained 7 dominant bands in DGGE gel were cloned and sequenced. 【Result】After cloning, nine sequences were obtained, of which 5 were archaea and 4 were not. Three sequences amongst 5 archaea showed 99% homology with Methanobrevibacter, and one sequence showed 98% of homology with Methanobrevibacter. The other sequence unclassified in Euryarchaeota showed 98% homology with Thermoplasmatales. In addition, four non archaea sequences were belonged to Bacteroidetes Firmicutes and Proteobacteria, but their homologies were relative less. 【Conclusion】DGGE technique was found useful in analyzing the diversity of rumen methanogens in buffalos.