Biology Department, Federal University of Lavras, Lavras, MG, Brazil.
Lett Appl Microbiol. 2010 Aug;51(2):131-7. doi: 10.1111/j.1472-765X.2010.02868.x. Epub 2010 May 14.
To develop species-specific primers capable of distinguishing between three important yeast species in alcoholic fermentation: Saccharomyces bayanus, Saccharomyces cerevisiae and Saccharomyces pastorianus.
Two sets of primers with sequences complementary to the HO genes from Saccharomyces sensu stricto species were used. The use of the ScHO primers produced a single amplificon of c. 400 or 300 bp with species S. cerevisiae and S. pastorianus, respectively. The second pair of primers (LgHO) was also constructed, within the HO gene, composed of perfectly conserved sequences common for S. bayanus species, which generate amplicon with 700 bp. No amplification product was observed in the DNA samples from non-Saccharomyces yeasts. Saccharomyces species have also been characterized via electrophoretic karyotyping using pulsed-field gel electrophoresis to demonstrate chromosomal polymorphisms and to determine the evolutionary distances between these species.
We conclude that our novel species-specific primers could be used to rapidly and accurately identify of the Saccharomyces species most commonly involved in fermentation processes using a PCR-based assay.
The method may be used for routine identification of the most common Saccharomyces sensu stricto yeasts involved in industrial fermentation processes in less than 3 h.
开发能够区分酒精发酵中三种重要酵母的种特异性引物:贝酵母、酿酒酵母和巴氏酵母。
使用与酿酒酵母种的 HO 基因互补的两套引物。使用 ScHO 引物可分别产生约 400 或 300 bp 的单扩增子,用于鉴定酿酒酵母和巴氏酵母。第二对引物(LgHO)也构建在 HO 基因内,由贝酵母种共有的完全保守序列组成,产生约 700 bp 的扩增子。在非酿酒酵母的 DNA 样本中未观察到扩增产物。还使用脉冲场凝胶电泳的电泳核型分析对酿酒酵母种进行了特征描述,以证明染色体多态性,并确定这些种之间的进化距离。
我们得出结论,我们的新型种特异性引物可以通过基于 PCR 的检测方法,快速准确地鉴定发酵过程中最常见的酿酒酵母种。
该方法可在不到 3 小时的时间内用于常规鉴定工业发酵过程中最常见的酿酒酵母种。