陈化年份与真菌催陈对广陈皮生物活性物质及真菌群落结构的影响

Study on Changes of Volatile Flavor Substances and Microbial Communities in Citri Reticulatae Pericarpium Under Different Years and Microbial Inoculation

摘要:

为探究不同陈化年份及真菌菌株对广陈皮生物活性物质及真菌组成影响，以陈化0、1、2、3、8、16年陈皮以及接种了Cladosporium perangustum CP16-M1、Aspergillus versicolor CP16-M2、Penicillium brocae CP16-M4的陈皮为材料，采用GC-MS检测与高通量测序等方法，测定分析陈皮总黄酮、挥发性物质、表面真菌“属”水平分布特点。结果显示，陈化8年是总黄酮积累关键期；挥发性成分以萜烯D-柠檬烯为主，以及醇类、酚类等共40种。16年陈皮中挥发性成分共30种，其中的L -香芹酮（含量2.36%）可能是老陈皮“薄荷香”的物质基础。接种CP16-M2、CP16-M4后总黄酮较对照组显著提升（P < 0.05）；接种菌株增加挥发性成分种类，CP16-M4使广陈皮特有成分N-甲基邻氨基苯甲酸甲酯含量上升至37.91%。真菌组成方面，Xeromyces丰度随陈化时间而增加，陈化16年时丰度达95.82%；新制陈皮优势真菌为Clavispora（50.37%）与其他年份差异最大。接种CP16-M1、CP16-M2、CP16-M4后，优势菌属分别为Hanseniaspora（44.54%）、Clavispora（34.74%）、Xeromyces（33.35%）。Penicillium，Aspergillus分别与D-柠檬烯，3-蒈烯呈强正相关（P < 0.001）。综上，陈皮挥发性成分与真菌组成存在相关性；菌株的接种可改变陈皮真菌丰度。本研究为解析陈化机制、定向调控品质提供理论支撑。

Abstract:

To investigate the effects of different aging years (0, 1, 2, 3, 8, and 16 years) and inoculation with three fungal strains on the bioactive substances and microbial community of Citri Reticulatae Pericarpium, the strains used were Cladosporium perangustum CP16-M1, Aspergillus versicolor CP16-M2, and Penicillium brocae CP16-M4.GC-MS and high-throughput sequencing were used to analyze total flavonoids, volatile compounds, and quality-related microorganisms in the samples. The results showed that the 8-year-aged sample was a critical period for total flavonoid accumulation. Terpene D-limonene was the main volatile component, and a total of 40 compounds including alcohols and phenols were identified.The 16-year-aged sample contained 30 volatile components, among which L-carvone (2.36%) might be the material basis for the "minty aroma" of aged Citri Reticulatae Pericarpium. After inoculation with CP16-M2 and CP16-M4, the total flavonoid content was significantly higher than that in the control group (P < 0.05). Inoculation with the three strains increased the number of volatile components, and CP16-M4 increased the content of methyl N-methylanthranilate—a unique component of Citri Reticulatae Pericarpium to 37.91%..The abundance of Xeromyces increased with aging time, reaching 95.82% in the 16-year-aged sample. Clavispora, the dominant fungus in the fresh sample (50.37%), showed the greatest difference from samples of other years. After inoculation with the three strains, the dominant genera were Hanseniaspora (44.54%), Clavispora (34.74%), and Xeromyces (33.35%), respectively. Penicillium and Aspergillus showed a strong positive correlation with D-limonene and 3-carene, respectively (P < 0.001). In conclusion, there is a correlation between the volatile components and microbial community. Inoculation with the three strains isolated from 16-year-aged Citri Reticulatae Pericarpium can alter the microbial community, providing theoretical support for elucidating the aging mechanism and directional regulating the quality.