食品科学 ›› 2012, Vol. 33 ›› Issue (15): 195-200.

• 生物工程 • 上一篇    下一篇

几株酵母菌的分子系统学鉴定

王泽举,王汝瑱,孙 悦,杨 莹,刘延琳*   

  1. 西北农林科技大学葡萄酒学院,陕西省葡萄与葡萄酒工程技术研究中心
  • 收稿日期:2011-07-01 修回日期:2012-07-19 出版日期:2012-08-15 发布日期:2012-09-07
  • 通讯作者: 刘延琳 E-mail:yanlinliu@nwsuaf.edu.cn
  • 基金资助:

    国家葡萄产业技术体系项目

Molecular Phylogenic Identification of Wine-Related Yeasts

  • Received:2011-07-01 Revised:2012-07-19 Online:2012-08-15 Published:2012-09-07
  • Contact: Yanlin Liu E-mail:yanlinliu@nwsuaf.edu.cn

摘要: 通过对酵母菌5.8S核糖体RNA的ITS区域PCR扩增及限制性酶切片段多态性(RFLP)的图谱分析以和26S rDNA D1/D2区及5.8S-ITS区的序列分析,共鉴定13株分离自新疆鄯善地区的葡萄酒相关酵母菌。5.8S-ITS区的4种内切酶(HaeⅢ、Hin6Ⅰ、HinfⅠ、AluⅠ)的酶切分析产出4种特异性图谱。根据26S rDNA D1/D2区的序列分析和BLAST比对,将供试菌株鉴定为4个属4个种,分别为Saccharomyces cerevisiae、Candida zemplinina、Hanseniaspora uvarum、Pichia kluyveri var. kluyveri;而根据5.8S-ITS-RFLP及5.8S-ITS序列分析,将供试菌种鉴定为Saccharomyces cerevisiae、Candida zemplinina、Hanseniaspora uvarum、Issatchenkia terricola 4个属4个种。3种方法对供试的10个菌株的鉴定结果一致,而对另外3株的鉴定结果不同。

关键词: 葡萄酒相关酵母, 分子系统学, 鉴定, 5.8S-ITS-RFLP, 26S rDNA D1/D2序列分析, 5.8S-ITS序列分析

Abstract: In this study, thirteen wine-related wild yeast strains from Shanshan, Xijiang were identified by PCR-RFLP (restriction fragment length polymorphism) and sequence analysis of the 5.8S-ITS region and the 26S rDNA D1/D2 domains. The PCR amplified 5.8S-ITS region were digested with four restriction endonucleases, Hae Ⅲ, Hin6Ⅰ, HinfⅠand AluⅠ to obtain 4 specific patterns. According to the sequence analysis of the 26S rDNA D1/D2 domains and BLAST alignment, the 13 yeast strains were identified as Saccharomyces cerevisiae, Candida zemplinina, Hanseniaspora uvarum and Pichia kluyveri var. kluyveri, while they were identified as Saccharomyces cerevisiae, Candida zemplinina, Hanseniaspora uvarum and Issatchenkia terricola according to the PCR-RFLP and sequence analysis of the 5.8S-ITS region. The same identification results for 10 of the 13 isolated yeast strains were obtained using the three methods, whereas inconsistent identification results were obtained for other three.

Key words: wine-related yeasts, molecular phylogeny, identification, 5.8S-ITS-RFLP, 26S rDNA D1/D2 sequence, 5.8S-ITS sequence

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