红曲黄酒传统酿造体系中真菌菌群多样性分析方法的构建及其应用  

Establishment and Application of the Analysis Methods of Fungal Diversity of Traditional Fermentation Starters from Hong Qu in Glutinous Rice Wine

作　　者：吕旭聪[1,2] 刘志彬 张雯[1] 陈芳[1] 黄若兰[1] 翁星[1] 饶平凡[1] 倪莉[1] LU Xucong;Liu Zhibin;Zhang Wen;Chen Fang;Huang Ruolan;Weng Xing;Rao Pingfan;Ni Li(Institute of Food Science and Technology,Fuzhou University,Fujian Center of Excellence for Food Biotechnology,Fuzhou 350108;National Engineering Research Center of JUNCAO Technology,College of Life Science,Fujian Agriculture and Forestry University,Fuzhou 350002)

机构地区：[1]福州大学食品科学技术研究所福建省食品生物技术创新工程技术研究中心,福州350108 [2]福建农林大学生命科学学院国家菌草工程技术研究中心,福州350002

出　　处：《中国食品学报》2018年第8期214-223,共10页Journal of Chinese Institute Of Food Science and Technology

基　　金：国家自然科学基金项目(31371820;31601466)

摘　　要：基于18S rRNA基因序列的PCR-RFLP分型方法分析前期从传统酒曲中分离纯化的16株代表性真菌,发现采用Hinf Ⅰ、Hae Ⅲ和Taq Ⅰ 3种限制性内切酶可将不同类型的真菌区分开,甚至是物种极为接近的真菌菌株。比较不同PCR引物(NS1/FR1+、FF390/FR1+、NS1/GCfung和NS3+/YM951r)对不同真菌菌株的扩增效率及DGGE分离效果的影响,筛选出最适合于红曲黄酒真菌菌群分析的PCR引物——NS1/GCfung。基于18S rDNA克隆文库RFLP分型结合克隆子测序技术解析红曲黄酒传统酿造体系中的真菌菌群结构,采用引物NS1/GCfung进行PCR-DGGE分析红曲黄酒传统酿造体系中的真菌菌群动态变化,鉴定出传统酒曲以及传统酿造过程中的优势真菌,为全面解析红曲黄酒传统酿造机制,改进传统酿造工艺,提升产品品质等奠定基础。In this study, 16 representative fungal strains isolated from traditional fermentation starters were analyzed by PCR-RFLP based on 18S rRNA gene sequence. The results showed that the three restrictive endonuclease Hinf I, Hoe III and Taq I can effectively differentiate different types of fungi, and even fungal strains whose relationship are relatively close. On the other hand, the effects of different PCR primers （NS1/FRI＋, FF390/FRl＋, NS1/GCfung andNS3＋/YM951r） on the amplification efficiency and DGGE separation efficiency of different fungal strains were compared. According to the results of PCR amplification, three primer sets （FF390/FRl＋, NS1/GCfung, NS3＋/YM951r） seemed to be suitable for the investigation of fungal community structure during the traditional brewing of Hong Qu glutinous rice wine. Of which, NS1/GCfung is the most suitable primer set for PCR-DGGE analysis of fungal community structure in traditional fermentation starters for Hong Qu glutinous rice wine. Finally, the fungal flora structures during the traditional brewing of Hong Qu glutinous rice wine were analyzed based on 18S rDNA clone library RFLP typing and cloning sub- sequencing technology. Besides, PCR-DGGE technology based on NS1/GCfung was also used to analyze the fungal diver- sity in the traditional brewing system of Hong Qu glutinous rice wine. The results showed that the traditional brewing process could be improved and the product quality could be improved.

关 键 词：红曲黄酒 酒曲 真菌多样性 PCR-RFLP PCR-DGGE 

分 类 号：TS262.4[轻工技术与工程—发酵工程]