中国葡萄酒产区酒酒球菌种质资源遗传多样性分析

doi:10.7506/spkx1002-6630-201517026

食品科学

中国葡萄酒产区酒酒球菌种质资源遗传多样性分析

金刚1，王华1,2,3，张昂4，李华1,2,3,*

1.西北农林科技大学葡萄酒学院，陕西杨凌 712100；2.西北农林科技大学，合阳葡萄试验示范站，陕西合阳 715300；
3.陕西省葡萄与葡萄酒工程技术研究中心，陕西杨凌 712100；4.秦皇岛出入境检验检疫局，河北秦皇岛 066004

出版日期:2015-09-15 发布日期:2015-09-11

Genetic Diversity Analysis of Oenococcus oeni Strains Isolated from Chinese Wines

JIN Gang1, WANG Hua1,2,3, ZHANG Ang4, LI Hua1,2,3,*

1. College of Enology, Northwest A&F University, Yangling 712100, China; 2. Heyang Experimental Demonstration Station,
Northwest A&F University, Heyang 715300, China; 3. Shaanxi Engineering Research Center for Viti-Viniculture,
Yangling 712100, China; 4. Qinhuangdao Entry-Exit Inspection and Quarantine Bureau, Qinhuangdao 066004, China

Online:2015-09-15 Published:2015-09-11

摘要/Abstract

摘要：

为了解我国葡萄酒产区酒酒球菌种质资源遗传多样性，对22 株筛选自我国不同葡萄酒产区的乳酸细菌进行了种特异性聚合酶链式反应（species-specific polymerase chain reaction，PCR）分析、16S rRNA序列分析和扩增片段长度多态性分析（amplified fragment length polymorphism，AFLP）基因分型。种特异性PCR和16S rRNA序列分析表明22 株分离株为酒酒球菌（Oenococcus oeni）。建立了O. oeni基于HindⅢ和MseⅠ为内切酶的AFLP分析体系，对16 对引物组合进行了筛选，结果表明HT-MA、HT-MT、HT-MC、HG-MA、HG-MT、HC-MT为O. oeni AFLP分析的最佳引物组合，且实验重复性在98%以上。对O. oeni的AFLP分析结果显示：22 株O. oeni分为3 个簇群，且簇群间遗传相似性系数较小。所以，以HindⅢ和MseⅠ为内切酶的AFLP技术是研究O. oeni基因分型的有效方法，我国葡萄酒产区的O. oeni具有丰富的遗传多样性，O. oeni菌株间的遗传相似性系数不仅仅与其生态地理分布有关，可能还与其所处的微生态和其他因素有关。

关键词: 酒酒球菌, 种特异性PCR, 16S rRNA序列分析, 扩增片段长度多态性分析技术

Abstract:

Oenococcus oeni (O. oeni) is considered as the key processer of malolactic fermentation (MLF), which will
influence wine quality. Indigenous microorganisms are increasingly considered as one of the factors influencing the quality
of individual wines. In order to understand the genetic diversity of O. oeni, species-specific PCR and 16S rRNA sequence
were used to identify 22 bacterial strains isolated from wines from different producing regions in China. Amplified fragment
length polymorphism (AFLP) based on HindIII and Mse I was developed to analyze the genotypes of 22 O. oeni strains. The
results of species-specific PCR and 16S rRNA sequence analysis showed that the 22 isolates were O. oeni. Double restriction
enzyme digestion and ligation system, PCR reaction system, polyacrylamide gel electrophoresis were developed to analyze
O. oeni. The combinations of 16 AFLP primers were tested. The results showed that HT-MA, HT-MT, HT-MC, HG-MA,
HG-MT, and HC-MT were the optimal primer combinations for analyzing O. oeni. The 22 O. oeni strains were analyzed
by AFLP, and cluster analysis indicated that the 22 O. oeni strains fell into three groups and their genetic similarity
was low. In conclusion, AFLP is a good method to genotype O. oeni by using HindIII and MseI. The O. oeni strains in
China have very rich genetic diversity. The genetic similarity of O.oeni strains is related to their ecological geographic
distribution and other factors.

Key words: Oenococcus oeni, species-specific polymerase chain reaction, 16S rRNA sequence, amplified fragment length polymorphism (AFLP)

中图分类号:

TS261.1

金刚，王华，张昂，李华. 中国葡萄酒产区酒酒球菌种质资源遗传多样性分析[J]. 食品科学, doi: 10.7506/spkx1002-6630-201517026.

JIN Gang, WANG Hua, ZHANG Ang, LI Hua. Genetic Diversity Analysis of Oenococcus oeni Strains Isolated from Chinese Wines[J]. FOOD SCIENCE, doi: 10.7506/spkx1002-6630-201517026.

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中国葡萄酒产区酒酒球菌种质资源遗传多样性分析

Genetic Diversity Analysis of Oenococcus oeni Strains Isolated from Chinese Wines

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