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      不同品种雪茄烟叶发酵前后叶面微生物及化学成分变化

      Changes in Phyllosphere Microorganisms and Chemical Components of Different Cigar Tobacco Varieties Before and After Fermentation

      • 摘要: 雪茄烟叶品质受品种特性及发酵微生物的共同影响。本研究旨在解析不同品种雪茄烟叶发酵过程中叶面微生物群落结构特征及其与主要化学成分的关联性,为阐明品种特色形成的微生物学机制提供依据。以优质雪茄烟品种青雪107(QX)和云雪6号(YX)为试验材料,在恒温恒湿条件下进行发酵。采用高通量测序技术分析细菌16S rRNA和真菌ITS序列,结合主要化学成分测定,对比分析两品种烟叶发酵前后叶面微生物群落特征及其与化学指标的相关性。两品种发酵后叶面微生物群落结构差异显著。QX发酵后细菌与真菌群落Shannon指数显著升高、Simpson指数显著降低,新增Methylobacterium-Methylorubrum、不动杆菌属Acinetobacter和假单胞菌属Pseudomonas为优势细菌,桑帕约氏酵母属Sampaiozyma成为优势真菌;而YX发酵后细菌群落多样性降低,葡萄球菌属Staphylococcus相对丰度显著上升至绝对优势地位,其次是短波单胞菌属Brevundimonas,真菌仍以曲霉属Aspergillus和节担菌属Wallemia为主。随机森林分析进一步揭示,YX在发酵前后均具有特征细菌属假单胞菌属,而QX在发酵后拥有更多特征真菌属如Talaromyces等。共现网络分析显示,葡萄球菌属与曲霉属分别为两品种细菌和真菌群落的共有拮抗中心,而Methylobacterium-Methylorubrum和短波单胞菌属分别为QX和YX的协同中心,反映了品种特异的微生物互作模式。化学成分分析表明,发酵后YX的钾、氯、还原糖、淀粉、总氮及蛋白质含量均显著高于QX,仅烟碱含量低于QX。相关性分析表明,还原糖含量与不动杆菌属显著正相关,与葡萄球菌属负相关;钾含量与短波单胞菌属显著正相关;蛋白质含量与鞘氨醇单胞菌属正相关,与短波单胞菌属负相关。可见,关键菌群在发酵中发挥重要功能,核心节点的差异导致群落功能分化和化学成分转化的品种差异。

         

        Abstract: The quality of cigar tobacco is co-determined by variety characteristics and fermentation-associated microbiota. This study aims to analyze the structure of the phyllosphere microbial community during the fermentation of different cigar tobacco varieties and its correlation with major chemical components. The findings will provide a theoretical basis for elucidating the microbiological mechanism of variety-specific flavor formation. Two high-quality cigar tobacco varieties Qingxue 107 (QX) and Yunxue 6 (YX) were fermented under constant temperature and humidity conditions. High-throughput sequencing of bacterial 16S rRNA and fungal ITS regions, combined with chemical components analysis, was employed to compare phyllosphere microbial communities before and after fermentation between the two varieties and their correlations with chemical indices. Phyllosphere microbial communities differed significantly between the two varieties post-fermentation. QX exhibited significantly elevated Shannon and reduced Simpson indices for both bacterial and fungal communities. Methylobacterium-Methylorubrum, Acinetobacter, and Pseudomonas emerged as dominant bacterial genera, with Sampaiozyma becoming the dominant fungal genus. By contrast, YX exhibited reduced bacterial diversity post-fermentation, with Staphylococcus becoming markedly dominant, followed by Brevundimonas. Fungal communities remained predominantly Aspergillus and Wallemia. Random forest analysis further revealed that Pseudomonas was a characteristic bacterial genus for YX both before and after fermentation, whereas QX possessed more characteristic fungal genera, such as Talaromyces, after fermentation. Co-occurrence network analysis indicated that Staphylococcus and Aspergillus served as shared antagonistic hubs for bacterial and fungal communities in both varieties, respectively. Conversely, Methylobacterium-Methylorubrum and Brevundimonas acted as synergistic hubs in QX and YX, respectively, reflecting variety-specific microbial interaction patterns. Chemical analysis showed that YX had significantly higher contents of potassium, chlorine, reducing sugar, starch, total nitrogen, and protein than QX post-fermentation, whereas its nicotine content was lower. Correlation analysis revealed that reducing sugar content was significantly positively correlated with Acinetobacter but significantly negatively correlated with Staphylococcus. Potassium content showed a significant positive correlation with Brevundimonas, while protein content was positively correlated with Sphingomonas but negatively correlated with Brevundimonas. These findings demonstrate that key microbial communities play crucial functional roles during fermentation, and differences in core microbial hubs drive variety-specific variations in community function and chemical transformation.

         

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