牦牛瘤胃中产纤维素酶芽孢杆菌的分离鉴定

doi:10.16431/j.cnki.1671-7236.2016.02.035

摘要/Abstract

摘要： 为了解牦牛瘤胃中产纤维素酶的芽孢杆菌种类,无菌采集10份成年麦洼牦牛瘤胃内容物样本,经80 ℃高温水浴20 min后分离得到耐高温芽孢菌.将分离菌株点种至羧甲基纤维素钠培养基上,使用刚果红染液染色筛选产纤维素酶菌株,扩增产酶菌株16S rRNA序列并测序,进行同源性比对和系统进化分析.结果显示:从10份样本中分离到64株菌,从中筛选出产纤维素酶芽孢杆菌共23株,其中蜡样芽孢杆菌16株,苏云金芽孢杆菌7株.系统进化分析结果显示,蜡样芽孢杆菌共分为两大支,有一株菌单独聚为一支,另外15株菌聚为一支,其中这一支又分为五小支,具有一定的遗传多样性;7株苏云金芽孢杆菌分布在一个大支上.该结果为了解牦牛瘤胃产纤维素酶芽孢杆菌的种类和开发牦牛专用微生态制剂奠定了试验基础.

关键词: 牦牛; 瘤胃; 芽孢杆菌; 纤维素酶; 16S rRNA; 系统进化分析

Abstract: To investigate the species of cellulase-producing Bacillus in yak rumen,10 samples of rumen content were aseptically collected from 10 adult Maiwa yaks to isolate the heat-resistant Bacillus by water bath at 80 ℃ for 20 min.The cellulase-producing strains were screened using the CMC-Na medium and Congo red staining.The 16S rRNA gene sequence of those cellulose-producing strains were amplified and sequenced.The results showed that 64 strains were isolated from the 10 samples.Total 23 strains were identified as cellulase-producing bacillus,including 16 strains of Bacillus cereus,7 strains of Bacillus thuringiensis.Furthermore,phylogenetic analysis showed that the 16 Bacillus cereus strains were clustered into two branches:One isolate was clustered into a branch alone,the other 15 isolates were clustered into a branch which clustered into 5 small branches,showing that there was certain genetic diversity in the isolates of Bacillus cereus.And all 7 Bacillus thuringiensis strains were clustered into a branch.Hence,the results layed the foundation of investigating the species of cellulase-producing Bacillus in yak rumen and developing probiotics special for yak.

Key words: yak; rumen; Bacillus; cellulase; 16S rRNA; phylogenetic analysis

中图分类号:

S852.61⁺6

蒋培培, 刘福音, 邹智坤, 周晏阳, 汤承. 牦牛瘤胃中产纤维素酶芽孢杆菌的分离鉴定[J]. , 2016, 43(2): 527-534.

JIANG Pei-pei, LIU Fu-yin, ZOU Zhi-kun, ZHOU Yan-yang, TANG Cheng. Isolation and Identification of Cellulase-producing Bacillus in Yak Rumen[J]. , 2016, 43(2): 527-534.

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