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草业学报 ›› 2017, Vol. 26 ›› Issue (6): 176-184.DOI: 10.11686/cyxb2016315

• 研究论文 • 上一篇    下一篇

西藏地区牦牛瘤胃中兼性厌氧纤维素降解菌的分离鉴定

李君风1, 原现军1, 董志浩1, Seare Tajebe Desta1, 陈雷1, 白晰1, 白云峰2, 邵涛1,*   

  1. 1.南京农业大学饲草调制加工与贮藏研究所,江苏 南京 210095;
    2. 江苏省农业科学院,江苏 南京 210014
  • 收稿日期:2016-08-19 修回日期:2016-10-09 出版日期:2017-06-20 发布日期:2017-06-20
  • 通讯作者: E-mail: taoshaolan@163.com
  • 作者简介:李君风(1988-),女,甘肃临夏人,在读博士。E-mail: ljf126ff@126.com
  • 基金资助:
    江苏省自主创新项目“以秸秆饲料化、基料化利用为核心的技术方案”[CX(15)1003-3]和江苏省自然基金青年基金项目“纤维素酶基因工程乳酸菌的构建及青贮表达效果研究”(BK20130694)资助

Isolation and identification of facultatively anaerobic cellulolytic bacterium in the rumen of Tibetan yaks (Bos grunniens)

LI Jun-Feng1, YUAN Xian-Jun1, DONG Zhi-Hao1, Seare Tajebe Desta1, CHEN Lei1, BAI Xi1, BAI Yun-Feng2, SHAO Tao1,*   

  1. 1.Institute of Ensiling and Processing of Grass, Nanjing Agricultural University, Nanjing 210095, China;
    2.Jiangsu Academy of Agricultural Science, Nanjing 210014, China
  • Received:2016-08-19 Revised:2016-10-09 Online:2017-06-20 Published:2017-06-20

摘要: 本研究旨在从西藏地区牦牛瘤胃中分离出具有分解纤维素能力的兼性厌氧菌,为青贮饲料微生物添加剂的研制提供支持。试验采集了西藏那曲地区成年牦牛的新鲜瘤胃液,经刚果红染色初筛,滤纸降解复筛,获得分解纤维素能力强的菌株,对菌株生长速率及酶活力进行测定,经形态学、生理生化特性和16S rRNA序列分析对菌株进行鉴定,并分析了菌株对水稻秸秆的降解效果。试验分离出一株兼性厌氧纤维素降解菌,属于肠球菌属,即粪肠球菌(Enterococcus faecalis) JF85。菌株JF85在15~55 ℃,pH 3.0~7.0及3.0%和6.5% NaCl培养基中生长良好。接种JF85,36 h后可获得最大内切葡聚糖酶活力(0.41 U/mL)和滤纸酶活力(0.13 U/mL)。在模拟发酵试验中,接种菌株JF85,14 d后水稻秸秆干物质、纤维素和半纤维素含量与对照组相比显著降低(P<0.05);在发酵第7天,JF85处理组显示最高的可溶性碳水化合物含量,整个发酵过程中木质素含量变化不显著(P>0.05)。菌株JF85具有分解纤维素、耐酸、耐盐特性,在青贮饲料生产中具有较好的应用前景。

Abstract: This study aimed to isolate and identify facultatively anaerobic cellulolytic bacterium in the rumen of yaks and to provide a basis for studying new types of silage additives. Fresh ruminal fluid was collected from Tibetan yaks and highly cellulolytic facultatively anaerobic bacteria were screened out by using Congo red staining and liquid secondary screening culture media. Cellulolytic bacteria strains were identified based on their morphological, physiological and biochemical characteristics, as well as by 16S rRNA analysis. Time course profile of growth and enzyme production by strain JF85 was obtained. The degradation efficiency of rice straw was examined by evaluating cellulose, hemicellulose and lignin contents under a range of condition. Strain JF85 was identified as Enterococcus faecalis. This strain grew well across a wide range of conditions: temperatures of 15-55 ℃, pH levels of 3.0-7.0 and NaCl levels of 3.0% and 6.5%. It exhibited the maximal endoglucanase activity (0.41 U/mL) and filter paper activity (0.13 U/mL) after 36 hours of incubation. In simulated fermentation, compared to the control, adding the JF85 strain significantly decreased the dry matter, cellulose and hemicellulose contents of rice straw after 14 days of inoculation (P<0.05). JF85-treated rice straw showed its highest water soluble carbohydrate content after 7 days of inoculation. However, there were no significant differences in lignin contents among all treatments. Applications of the JF85 strain, with its highly cellulolytic abilities and tolerance of acid and salt, have the potential to significantly improve the production of silage.