田间施药对自然发酵葡萄酒酵母菌群落结构的影响

Effect of pesticide application on the community structure of wine yeasts during spontaneous fermentation

摘要:

【背景】酵母菌是葡萄酒发酵过程中一类非常重要的微生物，其多样性及群体组成对葡萄酒的质量有重要贡献。影响葡萄酒中酵母菌组成的因素有很多，但目前尚未见葡萄园田管理对葡萄酒酵母菌群落结构影响方面的报道。【目的】探索田间施药对自然发酵葡萄酒酵母菌群落结构的影响。【方法】采用分离培养、常规分子生物学鉴定和Illumina MiSeq宏基因组测序结合的方法分析不同样品中的酵母菌群落结构情况。【结果】从不使用内吸收型化学农药的葡萄样品自然发酵液中分离鉴定出Pichia、Hanseniaspora、Schizosaccharomyces、Candida、Saccharomyces、Zygoascus、Issatchenkia等7个属8个种的酵母菌，宏基因组测序结果表明有Pichia (29.42%)、Saccharomyces (21.91%)、Issatchenkia (17.99%)、Hanseniaspora (12.10%)、Candida (7.47%)、Zygosaccharomyces (5.32%)、Schizosaccharomyces (3.07%)、Aureobasidium (0.29%)等属的酵母菌参与发酵；使用常规化学农药的葡萄样品自然发酵液中分离鉴定出Pichia、Hanseniaspora、Schizosaccharomyces、Candida、Cryptococcus等5个属6个种的酵母菌，宏基因组测序结果表明有Pichia (41.66%)、Hanseniaspora (21.54%)、Candida (19.11%)、Zygosaccharomyces (7.78%)、Schizosaccharomyces (4.04%)、Cryptococcus (3.21%)、Saccharomyces (1.12%)、Aureobasidium (0.49%)等属的酵母菌参与发酵。【结论】两样品中酵母菌比例有显著差异，表明在酿酒葡萄的园田管理中化学农药的使用对自然发酵葡萄酒的酵母菌群落结构有较大影响。

Abstract:

[Background] Yeast is a very important microorganism in wine fermentation. Its diversity and population composition play an important role in wine quality. There are many factors that affect the distribution and presence of yeast flora in wine. However, the influence of grape field management, for example the use of pesticides, on wine yeast community structure has not been reported yet. [Objective] We studied the effect of pesticide application on the community structure of wine yeasts during spontaneous fermentation. [Methods] Methods of pure culture, molecular biology identification and Illumina MiSeq metagenomic sequencing were used. [Results] In the spontaneous fermentation broth of grape samples without internal absorption chemical pesticides, 7 genuses (8 species) of yeast strains including Pichia, Hanseniaspora, Schizosaccharomyces, Candida, Saccharomyces, Zygoascus, and Issatchenkia were isolated and identified, while the result of metagenomic sequencing confirmed the dominance of Pichia (29.42%), Saccharomyces (21.91%), Issatchenkia (17.99%), Hanseniaspora (12.10%), Candida (7.47%), Zygosaccharomyces (5.32%), Schizosaccharomyces (3.07%), and Aureobasidium (0.29%). In the spontaneous fermentation broth of grape samples with conventional chemical pesticides, 5 genuses (6 species) of yeast strains including Pichia, Hanseniaspora, Schizosaccharomyces, Candida, and Cryptococcus were isolated and identified, while the result of Illumina MiSeq sequencing showed that Pichia (41.66%), Hanseniaspora (21.54%), Candida (19.11%), Zygosaccharomyces (7.78%), Schizosaccharomyces (4.04%), Cryptococcus (3.21%), Saccharomyces (1.12%), and Aureobasidium (0.49%) were involved in the fermentation. [Conclusion] Yeast community compositions in the two samples were significantly different, indicating that pesticide application on grapes had great influence on the community structure of wine yeasts during spontaneous fermentation.