Instituto Nacional de Tecnología Agropecuaria (INTA), EEA Mendoza and EEA La Consulta, San Martín 3853, 5507, Lujan de Cuyo, Mendoza, Argentina.
Curr Microbiol. 2010 Dec;61(6):506-14. doi: 10.1007/s00284-010-9645-y. Epub 2010 Apr 21.
Three molecular typing techniques were applied to assess the molecular relationships of Saccharomyces cerevisiae strains isolated from winery equipment, grapes, and spontaneous fermentation in a cellar located in "Zona Alta del Río Mendoza" (Argentina). In addition, commercial Saccharomyces strains widely used in this region were also included. Interdelta PCR typing, mtDNA restriction analysis, and microsatellite (SSR) genotyping were applied. Dendrograms were constructed based on similarity among different patterns of bands. The combination of the three techniques discriminated 34 strains among the 35 isolates. The results of this study show the complex relationships found at molecular level among the isolates that share the same ecological environment, i.e., the winemaking process. With a few exceptions, the yeast isolates were generally clustered in different ways, depending on the typing technique employed. Three clusters were conserved independently of the molecular method applied. These groups of yeasts always clustered together and had high degree of similarity. Furthermore, the dendrograms mostly showed clusters combining strains from winery and fermentation simultaneously. Most of the commercial strains included in this study were clustered separately from the other isolates analyzed, and just a few of them grouped with the strains mainly isolated from spontaneous fermentation. Only one commercial strain was clustered repetitively with a noncommercial strain isolated from spontaneous fermentation in the three dendrograms. On the other hand, this study has demonstrated the importance of selecting an appropriate molecular method according to the main objectives of the research.
三种分子分型技术被应用于评估来自阿根廷门多萨高地(“Zona Alta del Río Mendoza”)酿酒厂设备、葡萄和酒窖自然发酵的酿酒酵母菌株的分子关系。此外,还包括该地区广泛使用的商业酿酒酵母菌株。应用了 Interdelta PCR 分型、mtDNA 限制分析和微卫星(SSR)基因分型。基于不同带型模式之间的相似性构建了系统发育树。三种技术的组合将 35 个分离株中的 34 个菌株区分开来。这项研究的结果表明,在相同生态环境(即酿酒过程)中发现的分离株之间存在复杂的分子水平关系。除了少数例外,酵母分离株通常根据所采用的分型技术以不同的方式聚类。有三个聚类独立于所应用的分子方法被保留下来。这些酵母群总是聚集在一起,具有高度的相似性。此外,系统发育树大多显示出将酿酒厂和发酵同时分离的菌株组合在一起的聚类。本研究中包括的大多数商业菌株与分析的其他分离株分开聚类,只有少数与主要从自然发酵中分离的菌株聚类。只有一个商业菌株在三个系统发育树中与一个从自然发酵中分离的非商业菌株重复聚类。另一方面,这项研究表明了根据研究的主要目标选择适当的分子方法的重要性。