山羊瘤胃甲烷菌第七目菌株的富集、可利用底物及其主要共生菌的分析

发布时间：2018-09-06 06:41

【摘要】：在瘤胃中,甲烷菌第七目Methanomassiliicoccales(Mmc)是仅次于甲烷杆菌目的第二大菌群。Mmc菌株较难分离纯化,且尚未建立完善的系统进化分类体系。本文利用甲烷菌与产氢纤维降解细菌共分离的方法,富集纯化山羊瘤胃Mmc菌株,揭示其底物利用特性及共生细菌多样性。16S r RNA基因和mcr A基因鉴定结果显示:细菌培养液中仅含有一种Mmc甲烷菌J3。系统进化分析显示J3属于Mmc的"Ca.M.alvus"簇,与Candidatus Methanogranum caenicola相似性最高(96%)。与J3共生的细菌主要有Pyramidobacter piscolens(54.5%)、unclassified Ruminococcaceae(21.3%)、Blautia producta(14.8%)、Atopobium sp.(4.9%)、Eubacterium sp.(1.5%)。体外发酵结果显示,J3不但能够利用三甲胺,还能够利用甲醇。结论,与细菌共分离的方法能够在短期内从瘤胃液中富集纯化到Mmc甲烷菌。
[Abstract]:In rumen, the seventh order Methanomassiliicoccales (Mmc) of methanobacteria is the second largest group of methanobacterium, which is difficult to isolate and purify, and has not yet established a perfect phylogenetic classification system. In this paper, the method of co-isolation of methane bacteria and hydrogen-producing cellulolytic bacteria was used to enrich and purify goat rumen Mmc strain. The results showed that there was only one Mmc methanobacter J3in the culture medium of the bacteria. The results showed that there was only one Mmc methanobacter J3 in the culture medium. Phylogenetic analysis showed that J3 belonged to Mmc's "Ca.M.alvus" cluster and had the highest similarity with Candidatus Methanogranum caenicola (96%). The main bacteria symbiotic with J3 were Pyramidobacter piscolens (54.5%) unclassified Ruminococcaceae (21.3%) Blautia producta (14.8%) Atopobium sp. (4.9%) and Eubacterium sp. (1.5%). The results of in vitro fermentation showed that J3 could utilize not only trimethylamine but also methanol. Conclusion the method of co-isolation with bacteria can enrich and purify Mmc methanobacteria from rumen fluid in a short time.
【作者单位】：江苏省消化道营养与动物健康重点实验室/南京农业大学动物科技学院消化道微生物研究室;
【基金】：国家自然科学基金青年项目(31301999) 中央高校基本科研业务费自主创新重点研究项目(KYZ201412) 中央高校基本科研基金(KJ2013018)
【分类号】：Q93;S827

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