Use of specific PCR primers to identify three important industrial species of Saccharomyces genus: Saccharomyces cerevisiae, Saccharomyces bayanus and Saccharomyces pastorianus
Use of specific PCR primers to identify three important industrial species of Saccharomyces genus: Saccharomyces cerevisiae, Saccharomyces bayanus and Saccharomyces pastorianus
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DOI：
10.1111/j.1472-765x.2010.02868.x
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发表时间：
2010-08-01
影响因子：
2.4
通讯作者：
Schwan, R. F.
中科院分区：
文献类型：
--
作者：
de Melo Pereira, G. V.;Ramos, C. L.;Schwan, R. F.
目的：设计一对特异性引物，用于区分贝酵母（Saccharomyces bayanus）、酿酒酵母（Saccharomyces cerevisiae）和巴斯德酵母（Saccharomyces pastorianus）3种重要的酒精发酵酵母。ScHO引物的使用产生了c的单个扩增子。400或300 bp与S. cerevisiae和S.分别为pastorianus。第二对引物（LgHO）也在HO基因内构建，由S. bayanus物种，产生700 bp的扩增子。在来自非酵母属酵母的DNA样品中未观察到扩增产物。酵母属物种也已通过电泳核型分析，使用脉冲场凝胶电泳，以证明染色体多态性，并确定这些species.Conclusions之间的进化距离的特点：我们的结论是，我们的新的物种特异性引物可用于快速，准确地鉴定酵母属物种最常见的参与发酵过程中使用基于PCR的测定。该方法可用于在较短的时间内对工业发酵过程中最常见的严格酵母菌进行常规鉴定3小时以上。
Aim:To develop species-specific primers capable of distinguishing between three important yeast species in alcoholic fermentation: Saccharomyces bayanus, Saccharomyces cerevisiae and Saccharomyces pastorianus.Methods and Results: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.Conclusions: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.Significance and Impact of the Study: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.