应用传统的普洱熟茶渥堆方法进行发酵,研究分析普洱茶渥堆发酵过程中嗜热微生物的种类,应用测序技术对分离的微生物进行了鉴定。每天取样,进行微生物的分离、纯化及鉴定,并对分离出来的几种嗜热霉菌在不同温度、不同pH条件下培养,观察其生长情况。对普洱茶发酵过程中的微生物进行全面分析,运用分子生物学方法对分离的微生物进行鉴定。发现了普洱茶渥堆发酵过程中起重要作用的嗜热微生物,为揭示普洱茶高温发酵的机理奠定了基础。
Thermophilic fungi were isolated and identificated by the method of sequencing during traditional fermentation of puer tea. Sampling were carried out every day. Many types of microorganisms including thermophilic fungi were purified and Identified. Isolated thermophilic fungi were cultured at different temperatures and different pH, the growth state of thermophilic microorganisms were observed. This article showed a comprehensive analysis of microorganisms during the fermentation of puer tea, and identified the isolated microorganisms using molecular biology method. Thermophilic microorganisms which play an important role during the fermentation were discovered, it laid the foundation to reveal the mechanism of Puer tea fermentation at high temperature.
[1] 刘勤晋. 普洱茶之科学读本[M]. 广东: 广东旅游出版社, 2005: 26-27.
[2] 陈宗道, 周才琼, 童华荣. 茶叶化学工程学[M]. 重庆:西南师范大学出版社, 1999: 180-181.
[3] 赵龙飞, 周红杰. 云南普洱茶渥堆过程中的主要微生物初探[J]. 商丘师范学院报报, 2005, 21(2): 129-133.
[4] 董坤, 熊辛宇, 蓝增全. 普洱茶发酵过程中微生物类群分析[J]. 现代农业科技, 2009(1): 164-165.
[5] 刘勤晋, 周才琼, 许鸿亮, 等. 普洱茶的渥堆作用[J]. 茶叶科学, 1986, 6(2): 55-56.
[6] 何国藩, 林月蝉, 徐祥福. 广东普洱茶渥堆中细胞组织的显微变化及微生物分析[J]. 茶叶科学, 1987, 7(2): 54-57.
[7] 周红杰, 李家华, 赵龙飞, 等. 渥堆过程中主要微生物对云南普洱茶品质形成的研究[J]. 茶叶科学, 2004, 24(3): 212-218.
[8] 仓道平, 温琼英. 茯砖茶发酵中优势菌与有害菌类的分离鉴定[J]. 茶叶通讯, 1981(2): 12-14.
[9] 温琼英. 茯砖茶中主要微生物的研究[J]. 茶叶通讯, 1986(4): 19-21.
[10] 齐祖同, 孙曾美. 茯砖茶中优势菌群的鉴定[J]. 真菌学报, 1990, 9(3): 176-179.
[11] 陈可可, 朱宏涛, 王东, 等. 普洱熟茶后发酵加工过程中曲霉菌的分离和鉴定[J]. 云南植物研究, 2006, 28(2): l23-l26.
[12] Pitt J I, Hocking A D.Fungi and Food Spoilage[M]. Gaithersburg: Aspen Publishers Inc, 1999: 593.
[13] Samson RA, Hoekstra ES, Frisvad JC, et al. Introduction to Food and Airborne Fungi(6th edition)[M]. Utrecht: Centraalbureau voor Schimmelcultures, 2002: 389.
[14] 陈凤毛. 真菌ITS区序列结构及其应用[J]. 林业科技开发, 2007, 21(2): 5-7.
[15] Ramesh Maheshwari, Girish Bharadwaj, Mahalingeshwara K Bhat, et al. Thermophilic Fungi: Their Physiology and Enzymes[M]. Microbiology and Molecular Biology Reviews, 2000, 64(3): 461-488.