【摘要】：在瘤胃中,甲烷菌第七目Methanomassiliicoccales(Mmc)是僅次于甲烷桿菌目的第二大菌群。Mmc菌株較難分離純化,且尚未建立完善的系統(tǒng)進(jìn)化分類體系。本文利用甲烷菌與產(chǎn)氫纖維降解細(xì)菌共分離的方法,富集純化山羊瘤胃Mmc菌株,揭示其底物利用特性及共生細(xì)菌多樣性。16S r RNA基因和mcr A基因鑒定結(jié)果顯示:細(xì)菌培養(yǎng)液中僅含有一種Mmc甲烷菌J3。系統(tǒng)進(jìn)化分析顯示J3屬于Mmc的"Ca.M.alvus"簇,與Candidatus Methanogranum caenicola相似性最高(96%)。與J3共生的細(xì)菌主要有Pyramidobacter piscolens(54.5%)、unclassified Ruminococcaceae(21.3%)、Blautia producta(14.8%)、Atopobium sp.(4.9%)、Eubacterium sp.(1.5%)。體外發(fā)酵結(jié)果顯示,J3不但能夠利用三甲胺,還能夠利用甲醇。結(jié)論,與細(xì)菌共分離的方法能夠在短期內(nèi)從瘤胃液中富集純化到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.
【作者單位】： 江蘇省消化道營養(yǎng)與動物健康重點(diǎn)實(shí)驗(yàn)室/南京農(nóng)業(yè)大學(xué)動物科技學(xué)院消化道微生物研究室;
【基金】：國家自然科學(xué)基金青年項(xiàng)目(31301999) 中央高�；究蒲袠I(yè)務(wù)費(fèi)自主創(chuàng)新重點(diǎn)研究項(xiàng)目(KYZ201412) 中央高校基本科研基金(KJ2013018)
【分類號】：Q93;S827

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