Department of Biology and Biochemistry, University of Bath, Bath, UK.
FEMS Yeast Res. 2011 Nov;11(7):552-63. doi: 10.1111/j.1567-1364.2011.00745.x. Epub 2011 Aug 26.
The Saccharomyces genus (previously Saccharomyces sensu stricto) formally comprises Saccharomyces arboricola, Saccharomyces bayanus, Saccharomyces cariocanus, Saccharomyces cerevisiae, Saccharomyces kudriavzevii, Saccharomyces mikatae, Saccharomyces paradoxus and Saccharomyces pastorianus. Species-specific primer pairs that produce a single band of known and different product size have been developed for each member of the clade with the exception of S. pastorianus, which is a polyphyletic allopolyploid hybrid only found in lager breweries, and for which signature sequences could not be reliably created. Saccharomyces cariocanus is now regarded as an American variant of S. paradoxus, and accordingly a single primer pair that recognizes both species was developed. A different orthologous and essential housekeeping gene was used to detect each species, potentially avoiding competition between PCR primers and overlap between amplicons. In multiplex format, two or more different species could be identified in a single reaction; double and triple hybrids could not always be correctly identified. Forty-two unidentified yeasts from sugar cane juice fermentations were correctly identified as S. cerevisiae. A colony PCR method was developed that is rapid, robust, inexpensive and capable of automation, requires no mycological expertise on the part of the user and is thus useful for large-scale preliminary screens.
酿酒酵母属（以前称为酿酒酵母狭义种）正式包括酿酒酵母、贝酵母、卡氏酵母、酿酒酵母、克鲁维酵母、米曲霉、反常酵母和巴氏酵母。除了巴氏酵母，该属的每个成员都开发了具有特定引物对，这些引物对产生已知和不同大小的单一产物。巴氏酵母是仅在贮藏啤酒厂中发现的多倍体异源杂种，无法可靠地创建其特征序列。卡氏酵母现在被认为是反常酵母的一个美洲变体，因此开发了一种能够识别这两个物种的单一引物对。使用不同的直系同源和必需的管家基因来检测每个物种，从而潜在地避免了 PCR 引物之间的竞争和扩增子之间的重叠。在多重格式中，可以在单个反应中鉴定出两种或更多种不同的物种；但不能总是正确识别双杂交和三杂交。从甘蔗汁发酵中鉴定出的 42 种未鉴定酵母均被正确鉴定为酿酒酵母。开发了一种快速、稳健、廉价且可自动化的菌落 PCR 方法，不需要用户具备真菌学专业知识，因此非常适合大规模的初步筛选。
