瘤胃降解粗纤维产甲烷的厌氧真菌与甲烷菌共培养物的分离鉴定  

Isolation and identification of cellulolytic anaerobic fungi and their associated methanogens from Holstein Cow

作　　者：孙美洲[1] 金巍[1] 李袁飞[1] 毛胜勇[1] 成艳芬[1] 朱伟云[1] 

机构地区：[1]南京农业大学消化道微生物实验室,江苏南京210095

出　　处：《微生物学报》2014年第5期563-571,共9页Acta Microbiologica Sinica

基　　金：国家自然科学基金项目(31072052;31101735);中央高校基本科研业务费专项资金(KYZ201312)~~

摘　　要：【目的】本试验从瘤胃中分离鉴定降解粗纤维产甲烷的厌氧真菌与甲烷菌共培养物,为深入探究甲烷菌对厌氧真菌代谢途径的影响及相关调节机制奠定基础。【方法】利用厌氧滚管技术从荷斯坦奶牛瘤胃内容物中分离厌氧真菌与甲烷菌共培养物,通过形态学观察和DAPI染色以及甲烷菌16S rRNA基因序列分析方法分别对厌氧真菌及甲烷菌进行鉴定。【结果】从荷斯坦奶牛瘤胃中共分离到28株厌氧真菌与甲烷菌共培养物。共培养物中的厌氧真菌均为单中心菌株,分别属于Piromyces,Neocallimastix和Caeomyces属,所占百分比为53.57%,42.86%及3.57%。甲烷菌16S rRNA基因序列分析结果表明,共培养物中的甲烷菌均为甲烷短杆菌。本研究共获得四种不同的厌氧真菌与甲烷菌组合,分别为Piromyces/类Methanobrevibacter olleyae菌株,Neocallimastix/类Methanobrevibacter olleyae菌株,Neocallimastix/类Methanobrevibacter thaueri菌株及Caecomyces/类Methanobrevibacter olleyae菌株,分别占总数的53.57%,39.29%,3.57%及3.57%。【结论】分离得到的28株厌氧真菌和甲烷菌共培养物中,占优势的为具有丰富丝状假根的厌氧真菌Piromyces和Neocallimastix以及类Methanobrevibacter olleyae属的甲烷短杆菌。本研究为进一步研究瘤胃内厌氧真菌与甲烷菌相互代谢关系奠定基础。[ Objective] We studied the microbial interaction between anaerobic fungi and methanogens in the rumen of Holstein Cow. [ Methods ] Co-cultures of anaerobic fungi with indigenously associated methanogen were isolated by Hungate roll-tube technique. The anaerobic fungi were identified by morphology and 4＇ , 6 diamidino-2-phylindole nucleus staining and the methanogens were identified by 16S rRNA gene sequencing. [ Results] A total of 28 co-cultures of anaerobic fungus with indigenously associated methanogen were obtained. The anaerobic fungi in the co-cultures were identified as monocentric genera Piromyces, Neocallimastix and Caeomyces. The indigenously associated methanogens were Methanobrevibacter olleyae like and Methanobrevibacter thaueri like strains. Four different phylotypes of fungus-methanogen co-cultures were obtained, which were Piromyces/Methanobrevibacter olleyae like strains, Neocallimastix/ Methanobrevibacter olleyae like strains, Neocallimastix/Methanobrevibacter thaueri like strains and Caecomyces/ Methanobrevibacter oUeyae like strains. [ Conclusion] Our study isolated and identified 28 co-cultures of anaerobic fungus and associated methanogens, which provided new materials for further study the mechanism of methane emission in the rumen.

关 键 词：厌氧真菌 甲烷菌 共培养 

分 类 号：S852.6[农业科学—基础兽医学]