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基于挥发性组学与微生物组学的绿茶和红茶米酒品质差异研究

Differential Volatiles and Sensory Quality of Green Tea- and Black Tea-infused Rice Wines

  • 摘要:
    目的 探究绿茶和红茶添加对茶米酒风味品质、微生物群落演替及潜在功能活性的影响,为差异化风味发酵饮品开发提供科学依据。
    方法 以同源鲜叶制备的绿茶与红茶为基质,在相同酿造体系下制备绿茶米酒(W-GT)和红茶米酒(W-BT),并设空白对照(不加茶汤,以等量蒸馏水代替,W-C),对3组米酒样品进行感官审评。通过16S rRNA及ITS高通量测序解析细菌与真菌群落结构,利用PICRUSt2进行功能预测。运用顶空固相微萃取-气相色谱-质谱联用(headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry, HS-SPME-GC-MS)结合相对气味活度值(relative odor activity value, rOAV)鉴定特征风味组分,并整合微生物-代谢物关联分析揭示核心菌群对风味形成的调控作用。
    结果 感官审评结果表明,W-GT呈现清澈的黄绿色,滋味清爽且具幽雅清香;W-BT色泽橙黄明亮,口感醇厚饱满,蜜香显著。微生物组学分析显示,W-BT组的功能基因总丰度最高,尤其在代谢通路方面表现突出,表明其在代谢活性和功能多样性方面具有显著优势;W-GT组的功能基因则在遗传信息处理与耐药相关通路高度富集。16S rRNA在门水平上,厚壁菌门(p_Firmicutes)在W-GT组中占据主导地位;属水平上,W-BT与W-GT组均以明串珠菌属(g_Leuconostoc)为优势菌。ITS在门水平上,3组茶酒均以毛霉门(p_Mucoromycota)为优势菌;属水平上,酵母属(g_Saccharomyces)和毛霉属(g_Saccharomyces)在W-BT组中丰度最高,而W-GT组则展现出更高的真菌多样性。GC-MS共鉴定出922种挥发性化合物,以酯类、萜类、酮类为主。rOAV风味分析结果显示,3组米酒共有rOAV核心特征组分包括β-紫罗兰酮、3-甲基戊醛、(Z,Z)-3,6-壬二烯醛、(E,Z)-2,6-壬二烯醛、8-壬烯醛、(E)-2-己烯醛、(E,E)-3,5-辛二烯-2-酮、(E)-β-紫罗兰酮、反式,顺式-2,6-壬二烯-1-醇和1-壬烯-3-酮,其中β-紫罗兰酮的rOAV值为18173.7522879.45,在3组米酒中占绝对优势。差异代谢物的类别特征分析结果显示,W-GT以萜类物质显著上调为特征,W-BT则表现为酚类与醛类物质的富集。微生物-代谢物关联分析揭示,分枝杆菌属、Blautia等核心菌群分别参与了绿茶米酒和红茶米酒特征风味物质的形成。
    结论 本研究从“茶-菌-酒”互作角度阐释了绿、红茶米酒品质差异的形成机制,绿茶通过其活性成分抑制微生物代谢活性,同时驱动萜类风味物质的富集;红茶则通过发酵转化多酚、富集产香益生菌而促进酚类与醛类风味积累。

     

    Abstract:
    Objective Effects of rice wine fermented with green tea or black tea on the sensory quality, microbial community, and functional activities of the resulting products were studied.
    Method Rice wines were brewed with green tea (W-GT) or black tea (W-BT) made from a same batch of fresh leaves, as well as control (W-C) without the tea addition but an equal volume of distilled water. On the prepared wines, sensory evaluation was conducted by an expert panel, microbe structures analyzed by 16S rRNA and ITS high-throughput sequencings, and functional activities predicted using PICRUSt2. The headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry (HS-SPME-GC-MS) was employed to identify characteristic flavor components with relative odor activity value (rOAV) determined. Correlation between the microbes and metabolites was used to unveil microbial roles in the wine flavor formation.
    Result The sensory panel indicated that the yellowish-green W-GT had a refreshing taste with a delicate, elegant green-tea aroma, while the bright orange-yellow W-BT exhibited a mellow flavor with a pronounced honey-like aroma and full-bodied mouthfeel. W-BT was abundant on the functional genes, particularly those in the metabolic pathways, whereas W-GT was highly enriched in the functional genes related to genetic information processing and drug resistance. Based on 16S rRNA, Firmicutes was the dominant phylum in W-GT, and Leuconostoc the prevailing genus in both W-BT and W-GT. The ITS sequencing showed that Mucoromycota dominated at the phylum level among all, Saccharomyces and Mucor abundant at the genus level in W-BT, and W-GT rich in fungal diversity. A total of 922 volatiles, mainly esters, terpenoids, and ketones, were identified by GC-MS. The top 10 characteristic components scored on rOAV were β-ionone, 3-methylpentanal, (Z,Z)-3,6-nonadienal, (E,Z)-2,6-nonadienal, 8-nonenal, (E)-2-hexenal, (E,E)-3,5-octadien-2-one, 1-Nonen-3-one, (E)-β-ionone, and trans, cis-2,6-nonadien-1-ol. Among them, β-ionone was significantly higher on the values ranging from 18,173.75 to 22,879.45. On differential metabolites, W-GT was significantly upregulated on terpenoids, while W-BT enriched on phenols and aldehydes. The correlations between the core microbiota of Mycobacterium in W-GT and Blautia in W-BT were involved in the formation of the characteristic flavor compounds in the two tea-infused rice wines.
    Conclusion From the perspective of tea–microbe–wine interactions, the cause of quality differentiations of W-GT and W-BT seemed apparent. The microbes from the added green tea regulated the metabolic activity that enriched the terpenoid formation in W-GT; whereas the polyphenols from the fermented black tea and the aldehydes formed from the aroma-producing probiotics augmented the opulent flavor in W-BT.

     

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