水牛瘤胃内热原体属甲烷菌的FISH－FACS分离技术构建及其代谢功能分析

项目名称： 水牛瘤胃内热原体属甲烷菌的FISH－FACS分离技术构建及其代谢功能分析

项目编号： No.31460613

项目类型： 地区科学基金项目

立项/批准年度： 2015

项目学科： 畜牧学与草地科学

项目作者： 杨承剑

作者单位： 广西壮族自治区水牛研究所

项目金额： 50万元

中文摘要： 调控瘤胃中甲烷菌的组成与代谢过程，是减少反刍动物瘤胃甲烷生成的重要途径。瘤胃热原体目甲烷菌（又叫rumen cluster C,RCC)）占据着甲烷生成中甲基产物利用的生态位，是瘤胃甲烷的重要甚至是主要生产者。探明瘤胃RCC甲烷菌生理代谢功能特点是调控瘤胃甲烷菌的组成及其代谢过程的重要理论基础，对减少甲烷生成具有重要实际意义。本研究通过对水牛瘤胃内Rumen Cluster C(RCC)甲烷菌的富集培养，采用荧光原位杂交-荧光激活分选技术（FISH-FACS）分离瘤胃RCC，应用全基因组多重置换扩增技术（MDA）扩增基因组DNA并进行宏基因组测序、分析和基因注释，以及RCC甲烷菌对不同碳源利用方式等代谢功能多样性的研究，最终构建瘤胃RCC菌的分离方法体系，并阐明瘤胃RCC的代谢功能特点，进而为水牛甲烷菌群的系统研究提供技术支持，并为水牛瘤胃甲烷菌群的调控提供理论依据。

中文关键词： 瘤胃甲烷菌；Rumen；Cluster；C；分离；FISH-FACS；宏基因组学

英文摘要： Inhibition of methanogen in rumen of ruminants is the main way to decrease methane emission from ruminants. Rumen Cluster C methanogen which is a kind of methanogen belong to Thermoplasmatales occupied the ecological niche which use methyl products in rumen to produce methane; they are one of the important methane producer in rumen. The manipulation of ruminal methanogen composition and their metabolic process are based on the metabolism and physiological characteristics of RCC, which is very important for mitigation of methane emission. This project will enrich buffalo ruminal RCC methanogen by cultivation and isolated them by FISH-FACS technology, then use MDA techniques to amplifiy genomic DNA and analyze it with metagenomic methods, such as sequencing and gene annotation. At the same time, the metabolic function diversity of RCC, such as the utilization of RCC on different carbon source, will be investigated. At the end of this project, the methods of ruminal RCC methanogen separation and analysis system will be constructed, and the ruminal RCC methanogen characteristics will be explored, and this information can be used as technology support and theoretical guidance for further investigation and manipulation of buffalo methanogen.

英文关键词： Ruminal methanogen;Rumen Cluster C;Isolation;FISH-FACS;Meta-genomics