23 株酿酒酵母ISSR指纹图谱分析及SCAR标记的建立

目的：构建酿酒酵母菌株的简单重复序列间多态性指纹图谱数据库并建立序列特异性扩增区（sequencecharacterized amplified region，SCAR）标记技术，为酿酒酵母菌株的分类、遗传亲缘关系鉴定及菌种专利保护提供可靠的DNA分子标记技术依据。方法：在简单重复序列间多态性（inter-simple sequence repeat，ISSR）指纹数据分析基础上进行聚类分析并对菌种进行分类鉴定，同时将酿酒酵母菌株9号和15号中扩增获得的ISSR特异性DNA带转化为可以直接用于菌株快速鉴定的SCAR分子标记。结果：构建23 株酿酒酵母的ISSR指纹图谱，并在相似系数为0.85水平上将23 个供试菌株分为3大类，其中，1、2、4、7、15、16、17、19、20、21、23聚为第一类；10、11、12、13、14、18号菌株聚为第二类且10号和11号菌为同一菌株；3、5、6、8、9、22号菌聚为第三类且属于同一菌株。此外，利用所获得的2 个特异性条带成功转化为序列特异性扩增区分子标记。结论：在生产上酿酒酵母菌株遗传背景差异不大，常存在同物异名现象，而采用ISSR指纹及其SCAR分子标记技术快速鉴定酿酒酵母菌株在工业生产上具有重要意义。

ISSR Fingerprint Analysis and SCAR Marker of 23 Strains of Saccharomyces cerevisiae

Objective: To construct inter-simple sequence repeat (ISSR) DNA fingerprinting database of Saccharomyces cerevisiae
and to establish corresponding specific sequence characterized amplified region (SCAR) markers for use in the classification,
identification and analysis of the genetic relationship among S. cerevisiae strains and patent protection. Methods: On the basis of
ISSR DNA fingerprinting analysis, 23 Saccharomyces cerevisiae strains were clustered by UPGMA and their classification and
identification were conducted. Then two specific ISSR bands from strains 9 and 15 were converted into two SCAR markers for
rapid strain identification. Results: The 23 strains could be divided into three categories. Of these, category I included strains 1, 2, 4, 7, 15,
16, 17, 19, 20, 21 and 23, category II included strains 10, 11, 12, 13, 14 and 18 in which strains 10 and 11 were the same, and category Ⅲ
included strains 3, 5, 6, 8, 9 and 22 which belonged to the same strain with a similarity coefficient of 0.85. Furthermore, two specific bands
were converted into SCAR marker for strain identification. Conclusion: The development of ISSR fingerprints and SCAR markers for
rapid identification of Saccharomyces cerevisiae strains is significant for industrial production because there is no significant difference in
genetic background of Saccharomyces cerevisiae strains and it is often occurs that the same strains are called different names.