用DGGE和Real-Time PCR对低温沼气池中产甲烷古菌群落的研究  

Study on the Methanogenic Archaea Communities in Low Temperature Biogas Digester by Real-Time PCR and DGGE Methods

作　　者：王彦伟[1] 徐凤花[1] 阮志勇[2] 宋金龙[2] 王庆[3] 赵斌[3] 

机构地区：[1]东北农业大学资源与环境学院,哈尔滨150030 [2]中国农业科学院农业资源与农业区划研究所,北京100081 [3]华中农业大学生命科学技术学院,武汉430070

出　　处：《中国沼气》2012年第1期8-12,共5页China Biogas

基　　金：中央级公益性科研院所基本科研业务费专项资金项目(2007-21低温沼气发酵优良微生物菌株的筛选;鉴定;组合及其关键基因的克隆与表达);(2009-3沼气低温发酵促进剂的示范应用研究);现代牧业(集团公司)攻关项目(MF20100518)

摘　　要：利用DGGE及Real-Time PCR技术对不同地区低温沼气池中沼泥行进发酵前期、后期产甲烷古菌群落变化进行了研究。DGGE技术用于分析发酵前后产甲烷古菌优势群落的变化;Real-Time PCR技术用于分析发酵前期、后期样品产甲烷古菌的数量变化。结果表明,沼气低温发酵的过程中,不同沼泥样品发酵前期、后期产甲烷古菌的优势群落差异明显且数量上也存在较大差异;主要变化的产甲烷古菌的优势群落为甲烷粒菌属(Methanocor-pusculum),甲烷八叠球菌属(Methanosarcina),甲烷鬃毛菌属(Methanosaeta)。甲烷八叠球菌属(Methanosarcina)在黑龙江沼泥样品(A1)发酵后期成为优势群落,甲烷鬃毛菌属(Methanosaeta)在山东发酵前期(B1),后期样品(B2)及安徽发酵前期样品(C1)中占有绝对优势。产甲烷古菌的数量介于104～105个.mL-1之间,其中黑龙江样品中产甲烷菌数量最多,而安徽样品中产甲烷菌数量最少。The change of the methanogenic archaea communities and the dominant archaea in low temperature biogas digest- er were investigated by DGGE. The quantity changes of methanogenic archaea were analyzed by Real-Time PCR. The re- suits indicated that there were significant difference in dominant communities and the archaea quantity between earlier stage and later stage of fermentation in the low temperature biogas digester. The Methanocorpusculum, Methanosarcina and Meth- anosaeta were the dominant population in the digester. Methanosarcina was the dominant population in the later fermenta- tion for Heilongjiang sample, but the Methanosaeta was the dominant in both earlier and later fermentation stage for Shan- dong sample, and the earlier stage for Anhui sample. The quantity of Methanogenic archaea ranged between 10^4-10^5个·mL^-1, among which, the Heilongjiang sample had the highest and the Anhui sample had the lowest.

关 键 词：沼泥 产甲烷古菌 DGGE REAL-TIME PCR 

分 类 号：Q93[生物学—微生物学] S216.4[农业科学—农业机械化工程]