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基于聚合酶链式反应的新型酵母鉴定方法。

New PCR-based methods for yeast identification.

作者信息

Hierro N, González A, Mas A, Guillamón J M

机构信息

Departament de Bioquímica i Biotecnologia, Facultat d'Enologia, Unitat d'Enologia, Centre de Referència en Tecnologia d'Aliments (CeRTA), Universitat Rovira i Virgili, Ramón y Cajal, Tarragona, Spain.

出版信息

J Appl Microbiol. 2004;97(4):792-801. doi: 10.1111/j.1365-2672.2004.02369.x.

Abstract

AIMS

To characterize reference yeast strains and identify indigenous strains isolated from wine fermentations by PCR methods.

METHODS AND RESULTS

We compared several PCR techniques for yeast identification. We used oligonucleotide primers that are complementary to (i) intron splice sites, (ii) REP and (iii) ERIC elements to produce PCR fingerprints that display specific patterns between the different yeast species. These three techniques were used to characterize 41 reference yeast strains belonging to 15 different species and to identify 40 indigenous strains isolated from grape must and wine fermentations. Species-specific banding patterns were obtained with the three PCR-techniques with different degrees of intraspecific differentiation depending on the method. By comparing the PCR fingerprints of unknown isolates with those produced by reference strains, we identified yeast strains isolated from an industrial wine fermentation.

CONCLUSIONS

All three PCR techniques are rapid, reliable and simple methods of yeast identification. As far as we know, this is the first time that the primers designed for amplifying repetitive elements in bacteria have been successfully used in yeast.

SIGNIFICANCE AND IMPACT OF THE STUDY

Industry needs rapid, reliable and simple methods of yeast identification. The proposed PCR techniques will allow to achieve this objective.

摘要

目的

通过PCR方法对参考酵母菌株进行特征分析,并鉴定从葡萄酒发酵中分离出的本土菌株。

方法与结果

我们比较了几种用于酵母鉴定的PCR技术。我们使用与(i)内含子剪接位点、(ii)REP和(iii)ERIC元件互补的寡核苷酸引物来产生PCR指纹图谱,这些指纹图谱在不同酵母菌种之间呈现出特定模式。这三种技术用于对属于15个不同物种的41株参考酵母菌株进行特征分析,并鉴定从葡萄汁和葡萄酒发酵中分离出的40株本土菌株。根据方法不同,这三种PCR技术获得了具有不同种内分化程度的物种特异性条带模式。通过将未知分离株的PCR指纹图谱与参考菌株产生的指纹图谱进行比较,我们鉴定了从工业葡萄酒发酵中分离出的酵母菌株。

结论

所有三种PCR技术都是快速、可靠且简单的酵母鉴定方法。据我们所知,这是首次将设计用于扩增细菌中重复元件的引物成功应用于酵母。

研究的意义和影响

行业需要快速、可靠且简单的酵母鉴定方法。所提出的PCR技术将有助于实现这一目标。

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