23株酿酒酵母ISSR指纹图谱分析及SCAR标记的建立

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

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(sequence characterized amplifi ed 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。

Objective： To construct inter-simple sequence repeat（ISSR） DNA f ingerprinting database of Saccharomyces cerevisiae and to establish corresponding specif ic sequence characterized amplif ied region（SCAR） markers for use in the classif ication, identif ication and analysis of the genetic relationship among S. cerevisiae strains and patent protection. Methods： On the basis of ISSR DNA f ingerprinting analysis, 23 Saccharomyces cerevisiae strains were clustered by UPGMA and their classif ication and identif ication were conducted. Then two specif ic ISSR bands from strains 9 and 15 were converted into two SCAR markers for rapid strain identif ication. 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 coeff icient of 0.85. Furthermore, two specif ic bands were converted into SCAR marker for strain identif ication. Conclusion： The development of ISSR f ingerprints and SCAR markers for rapid identif ication of Saccharomyces cerevisiae strains is signif icant for industrial production because there is no signif icant difference in genetic background of Saccharomyces cerevisiae strains and it is often occurs that the same strains are called different names.