基于rDNA ITS-1序列酿酒酵母的系统分类学研究

为了准确快速地为酵母生产提供优良菌株,采用核糖体DNA ITS-1(Internal transcribed spacer 1)序列分析法,对27株酵母菌(其中24株为酿酒酵母属中的酿酒酵母(S.cerevisiae),其他3株分别为汉逊酵母属中的多形酵母(H.polymorph)、克鲁维酵母属中的乳清酵母(K.marxianus)和红酵母属中的粘质酵母(R.mucilaginosa))进行了系统分类学研究,提取这27株酵母菌的基因组DNA并进行PCR扩增,采用琼脂糖和非变性聚丙烯酰胺凝胶电泳检测,能将酵母菌株鉴定到属的水平;对PCR扩增序列进行克隆和测序,结果表明大多数酿酒酵母菌株之间的ITS-1序列有差异,主要表现在序列中的一段Ploy-T上,最少的有7个T,最多的有13个T。研究还用Phylip 3.6等DNA序列分析软件对不同ITS-1序列的菌株进行了系统发育分析,确定出不同菌株之间的亲缘关系,并纠正了长期认为是酿酒酵母的两株酵母菌。

Phylogenetic reclassification analysis of S.cerevisiae based on the internal transcribed spacer 1(ITS-1) sequence of rDNA

In order to supply high quality yeast strains rapidly and exactly in production,the number of 27 yeast strains are researched by analyzing phylogenetic position,24 yeast strains of which are S.cerevisiae,the others are H.polymorph,K.marxianus and R.mucilaginosa respectively.The internal transcribed spacer sequences 1,part of 18S rDNA and 5S rDNA of yeast strains are amplified by PCR after DNA isolation and primer design.The strains between genus can be identified by AGE,and part of species can be identified by PAGE.By cloning and sequencing of the ITS-1 sequences,there are differences between many S.cerevisiae,except that few of them are same completely.The most diversity are series of sequences of Poly-T between 7 and 13.The taxonomic relationships between them are indicated by Phylip 3.6 with the different ITS-1 sequences of S.cerevisiae strains.