Characterization of fungal community and dynamics during the traditional brewing of Wuyi Hong Qu glutinous rice wine by means of multiple culture-independent methods
Characterization of fungal community and dynamics during the traditional brewing of Wuyi Hong Qu glutinous rice wine by means of multiple culture-independent methods
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多种非培养方法传统酿造武夷红曲糯米酒过程中真菌群落和动态的表征
DOI：
10.1016/j.foodcont.2015.01.046
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发表时间：
2015-08-01
期刊：
影响因子：
6
通讯作者：
Ni, Li
中科院分区：
文献类型：
--
作者：
Lv, Xu-Cong;Cai, Qi-Qi;Ni, Li
本研究采用巢式PCR-变性梯度凝胶电泳（PCR- dgge）、18S核糖体RNA （rRNA）基因克隆文库和种特异性定量PCR （qPCR）等非培养方法，对武义红曲传统发酵过程中的真菌群落及其动态进行了研究。巢式PCR-DGGE和18S rRNA基因克隆文库显示，不同酿造阶段的优势菌群存在差异。随着发酵的进行，在酿酒早期检测到的部分真菌种类（如红曲霉、米根霉、吉列蒙毕赤酵母和纤维酵母菌）的相对比例呈下降趋势，而酿酒后期则成为优势菌种。本研究发现，PCR-DGGE指纹图谱和18S rRNA基因克隆文库具有相似的结果。然而，两种不同分子生物学方法检测真菌种类的差异表明，巢式PCR-DGGE和18S rRNA基因克隆文库相结合的方法比单独使用一种方法更能全面地了解真菌的动态。最后，对巢式PCR-DGGE和18S rRNA基因克隆文库检测到的优势真菌种类进行定量分析。结果表明，在酿造初期，米曲霉菌数量急剧增加并占主导地位，但从第10天到传统酿造结束，米曲霉数量持续下降。在传统酿造过程的后期，酿酒酵母在早期急剧增加，成为主要酵母菌种。本研究首次采用多元非培养方法对武夷红曲糯米酒传统酿造过程中的真菌群落和动态进行了详细评价，对科学认识酒的酿造机理，选择适宜的有益菌种，提高酒的品质具有重要价值。(C) 2015 Elsevier Ltd.版权所有。
The objective of this study was to investigate the fungal community and dynamics during the traditional fermentation of Wuyi Hong Qu (black-skin-red-koji) glutinous rice wine using culture-independent approaches based on nested PCR-denaturing gradient gel electrophoresis (PCR-DGGE), 18S ribosomal RNA (rRNA) gene clone libraries and species-specific quantitative PCR (qPCR). Both of nested PCR-DGGE and 18S rRNA gene clone libraries revealed that the predominant microorganisms varied in different brewing phases. The relative proportions of some fungal species (such as Monascus purpureus, Rhizopus oryzae, Pichia guilliermondii and Saccharomycopsis fibuligera) detected at the early brewing stage decreased as the fermentation progressed, while Saccharomyces cerevisiae became the dominant species at the later stage. The present study revealed that PCR-DGGE fingerprint and 18S rRNA gene clone libraries presented the similar results. Nevertheless, the differences of the fungal species detected by the two different molecular biological methods demonstrated that the combined approach of nested PCR-DGGE and 18S rRNA gene clone libraries would give a more comprehensive profile of the fungal dynamics than either alone. Finally, species-specific qPCR was also performed to quantify some dominant fungal species detected by nested PCR-DGGE and 18S rRNA gene clone libraries. Results showed that R. oryzae increased sharply and dominated during the early brewing period, but decreased continuously from day 10 to the end of the traditional brewing. S. fibuligera and P. guilliermondii were the dominant yeast species at early stage, while S. cerevisiae increased sharply during the early phase and became the primary species at the end of the traditional brewing process. This study provides the first detailed evaluation of fungal community and dynamics in Wuyi Hong Qu glutinous rice wine traditional brewing process using multiplex culture-independent methods, which would show great value in scientifically understanding of the brewing mechanism and be useful for the selection of the suitable and beneficial fungal strains to improve the wine quality. (C) 2015 Elsevier Ltd. All rights reserved.