DGGE法初步解析茯砖茶发花过程中真菌群落结构

Analysis of fungal community structure during the Fahua-fermentation process of Fuzhuan Brick Tea by DGGE technology

为研究茯砖茶发花过程中真菌群落结构和种类,对发花过程中不同时段真菌落18S rDNA高变区进行扩增,并对真菌变性梯度凝胶电泳（DGGE）图谱中条带进行克隆、测序和序列比对分析。结果表明：茯砖茶发花前后时段其优势菌不同,以发花过程的第10天为临界点,0～8 d和10～14 d,前后存在2个差异较大的真菌优势种群结构的演变,10天前以好干性酵母菌、毕赤酵母、假丝酵母为优势菌,10 d后以阿姆斯特丹散囊菌、好干性酵母菌为优势菌;18S rDNA高变区比对结果表明,茯砖茶发花过程中有好干性酵母、汉逊德巴利酵母、酿酒酵母、假丝酵母菌、热带假丝酵母、路德酵母、毕赤酵母、牧草红酵母、隐球酵母、阿姆斯特丹散囊菌、曲霉、毛霉、安大略假单胞菌、真皮毛孢子菌、青霉、白地霉、灰绿曲霉、蜡叶散囊菌等;比对结果表明存在3株好干性酵母菌和5株阿姆斯特丹散囊菌,说明优势菌存在多种生态类型。采用DGGE指纹图谱能更系统、更真实地反映茯砖茶发花发酵过程中真菌群落结构和多样性动态变化。

To explore fungal community structure and species composition in Fahua-fermentation process of Fuzhuan Briek Tea, the fungal variable region of 18S rDNA was amplified during dif- ferent periods of Fahua-fermentation process. Amplified products were separated by denaturing gradient gel electrophoresis （DGGE） and the bands were cloned, sequenced and aligned. The results show that two distinctive kinds of fungal communities, correlating to the fermentation peri- ods, 0-8 d and 10-14 d respectively, were found. The dominant fungi of the former band in- elude Wallemia muriae, Pichia kudriavzevii, Candida sp. , and that of the latter include Euroti- um amstelodarni and IV. muriae. The results of variable regions of 18S rDNA alignment showed that IV. sebi, IV. muriae, Debaryomyces hansenii, Saccharomyces cerevisiae, Candida sp., C. tropicalis, Lodderomyces sp. , P. kudriavzevii, Rhodotorula graminis, Rhizomucor pusillus, Cryptococcus sp. , E. amstelodami, Aspergillus penicillioides, R. miehei, C. ontarioensis, Tricho- sporon dermatis, Penicillium decumbens, Galactomyces geotrichum, A. glaucus, E. herbariorum were detected. The 18S rDNA alignment results also confirmed that there were 3 strains of IV. se- bi and 5 strains of E. amstelodami, displaying a wide variety of ecotypes within, the dominant fungi. This study for the first time shows a detailed soectrum of funzi durinz the Fahua-fermentationprocess. DGGE fingerprint is an important technique to analyze the comprehensive, detailed fun- gat community structure and diversity dynamics of Fuzhuan Brick Tea during this process.