Application of Multi Locus Sequence Typing to the analysis of the biodiversity of indigenous Saccharomyces cerevisiae wine yeasts from Lebanon
Application of Multi Locus Sequence Typing to the analysis of the biodiversity of indigenous Saccharomyces cerevisiae wine yeasts from Lebanon
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DOI：
10.1111/j.1365-2672.2006.02817.x
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发表时间：
2006-04-01
影响因子：
4
通讯作者：
Gaillardin, C
中科院分区：
文献类型：
--
作者：
Ayoub, MJ;Legras, JL;Gaillardin, C
目的：评价多位点序列分型技术(MLST)在葡萄酒酵母菌株多样性研究中的适用性。方法和结果：对代表看家基因、未知功能的开放阅读框(ORF)或基因间隔区的26个基因座的DNA片段进行了扩增和测序。对从黎巴嫩传统酿酒厂分离的65株酿酒酵母、商品葡萄酒菌株和亚洲分离株等84株酿酒酵母进行了7个基因座的检测。总的序列多样性为2.05%，由单核苷酸多态组成，其中60%为杂合态。对同一组菌株进行了6个多态位点微卫星分型和增量间扩增。结论：MLST图谱的聚类性明显区分了亚洲菌株群与黎巴嫩和欧洲商品菌株之间的亲缘关系。与微卫星或三角间分型(>；99%)相比，目前的MLST方案对近缘葡萄酒酵母的区分性较小(92.27%)。研究的意义和影响：MLST是一种高度可靠的相关性推断方法，有望用于葡萄酒酵母分型。
Aims: To assess suitability of Multi Locus Sequence Typing (MLST) for investigating the biodiversity of wine yeast strains. This method was compared with established ones like microsatellite analysis or amplification of genomic regions flanked by repeated (delta) elements.Methods and Results: DNA fragments were amplified and sequenced for 26 loci representing housekeeping genes, open reading frames (ORFs) of unknown functions or intergenic regions. A set of seven loci was tested on 84 Saccharomyces cerevisiae strains, including 65 strains isolated from traditional wineries in Lebanon, commercial wine strains and Asian isolates. An overall sequence diversity of 2.05% was observed, consisting of single nucleotide polymorphisms, 60% of them occurring in a heterozygous state. The number of polymorphic sites per locus varied between 4 and 14. The same set of strains was analysed by microsatellite typing on six polymorphic loci and by interdelta amplification.Conclusions: Clustering of MLST profiles clearly differentiated the Asian group of strains from Lebanese and European commercial strains that appear closely related. The current MLST scheme appears less discriminatory (92.27%) on closely related wine yeasts than microsatellite or interdelta typing (> 99%).Significance and Impact of the Study: MLST is a highly reliable method for relatedness inference and promising for wine yeast typing.