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中国农学通报 ›› 2012, Vol. 28 ›› Issue (6): 189-193.

• 生物技术科学 • 上一篇    下一篇

低温条件下牛粪沼气产甲烷菌多样性初步研究

何志刚 牛世伟 于涛   

  • 收稿日期:2011-08-23 修回日期:2011-10-08 出版日期:2012-02-25 发布日期:2012-02-25

Methanogenic Diversity in an Anaerobic Digester with Manure Feces in Low Temperature

  • Received:2011-08-23 Revised:2011-10-08 Online:2012-02-25 Published:2012-02-25

摘要:

为了在低温条件下调查牛粪沼气产气高峰期的产甲烷菌菌群结构。采用分子生物学方法构建16S rDNA基因文库,随机分析文库中60个克隆的16S rDNA基因序列,研究了牛粪沼气池发酵液中产甲烷菌的菌群结构。结果表明,在60个克隆中,18个属于甲烷八叠球菌属(Methanosarcina sp.)与Methanosarcina spAK-16菌株同源程度最高,相似性为99%。7个属于甲烷八叠球菌科(Methanosarcinales),15个属于甲烷鬃毛菌属(Methanosaeta)。利用MEGA 4.0软件分析60个克隆的16S rDNA序列,构建相应的系统进化树,结果显示,60个克隆子主要分为两大类群GroupA和GroupB。其中以属于甲烷八叠球菌属(Methanosarcina sp.)为主要类群,占全部克隆的42%,另一部分以甲烷鬃毛菌属(Methanosaeta)为主要类群,占25%。在整个发酵周期中到投料第18天达到产气最高峰。随着产气高峰期的过后,产气量逐渐下降。1个周期的总产气量504.91 mL 折合8 m3发酵罐产气量21.71 m3,其中甲烷气体总量为62%。分析低温(4℃)条件下以牛粪为主要发酵原料的沼气池产气高峰期的优势产甲烷菌群,结果发现甲烷八叠球菌属(Methanosaeta sp.)占总数的42%。

关键词: 叶片形态结构, 叶片形态结构

Abstract:

Low temperature conditions, cow dung methane gas production peak structure of methane-producing bacteria flora, using molecular biology 16S rDNA gene library, research dung biogas fermentation flora methanogens. Structure analysis of 60 randomLy cloned library 16S rDNA gene sequence, was found in 60 clones belonging to 18 genera (Methanosarcina sp.). With one of (Methanosarcina sp.)AK-16 strain of the highest degree of homology, similarity of 99%. 7 of which are Division Methanosarcinales Methanosarcina strains the highest degree of homology, similarity of 99%, 15 belong to the genus Methanosaeta strains of methane mane highest degree of homology, similarity of 99% use software to analyze 60 MEGA4.0 cloned 16S rRNA sequences to construct the corresponding phylogenetic the showed that was divided into two groups, namely Group A and Group B. Which belongs to genus Methanosarcina as Methanosarcina sp. major groups, accounting for 42% of all clones, the other part of the genus Methanosaeta mane of methane as the main group, accounting for 25%. Throughout the fermentation period in the first 18 days until the feed gas production reached its peak. With later after the peak of gas production, gas production decreased to 49 days of the decline in methane gas production also decreased 21%. A period equivalent to the total gas production 504.91 mL 8 m3 21.71 m3 fermentation gas production total of which 62% of methane gas. Low temperature (4℃) under the conditions of the fermentation of cow dung as the main raw material for biogas production peak period of the advantages of methanogenic bacteria, and found a Methanosarcina Methanosarcina sp. accounted for 42% of the total.