不同生长年限山豆根内生菌群落特征及其与药材主要有效成分的相关性分析

    Correlations of Endophytic Microbial Community with Main Active Components of Sophora tonkinens of Different Growth Years

    • 摘要:
      目的 探究桂产道地药材山豆根内生菌群落特征及其与药材主要活性成分的关联,明确影响药效成分的关键功能菌群,为建立基于微生物群落调控的山豆根优质栽培技术奠定基础。
      方法 采用高通量测序技术分析2年(R2)、4年(R4)、6年(R6)生山豆根内生细菌和真菌群落结构,结合高效液相色谱法(HPLC)测定生物碱类有效成分含量,运用Spearman相关性分析揭示菌群与成分间的关系。
      结果 随着种植年限的增加,山豆根内生细菌的丰富度和多样性均显著升高,而内生真菌的多样性和丰富度变化较小。内生优势细菌门均为变形菌门(Proteobacteria),优势真菌门为子囊菌门(Ascomycota)。优势细菌属因年限而异:R2和R4优势菌属均为产碱菌科未分类属(unclassified_f_Alcaligenaceae),丰度相对较高、分别为89.35% 和60.09%;R6优势菌属为假单胞菌属(Pseudomonas),相对丰度为52.20%。优势真菌属也随年限变化显著,其中R2优势菌属为拟折孢属(Minimelanolocus),R4优势菌属为柯达酵母属(Codinaea),R6优势菌属为未分类真菌属(unclassified_k_Fungi)。有效成分含量分析显示,R4山豆根中苦参碱含量为0.30 mg/g,显著高于R6(P<0.01);氧化苦参碱含量为16.26 mg/g,极显著高于R2和R6(P<0.001);金雀花碱和N-甲基金雀花碱在R6中的含量分别为0.99 mg/g和0.052 mg/g,均显著高于R4(P<0.01)。相关性分析表明,产碱菌科未分类属、假单胞菌属及泛菌属(Pantoea)等内生细菌与山豆根的有效成分含量显著相关(P<0.05),赤壳属(Dactylonectria)内生真菌对山豆根有效成分的积累也有显著影响(P<0.05)。
      结论 明确与山豆根有效成分密切相关的潜在内生细菌和真菌,揭示特定功能菌群在调控药材品质形成中的关键作用,为深入解析内生菌-宿主互作影响药材品质的机理及制定优质栽培策略提供了科学依据。

       

      Abstract:
      Objective To explore the community characteristics of endophyte in Sophora tonkinens and its correlation with the main active components of medicinal materials. To clarify the key functional bacteria that affect the pharmacodynamic components, and lay a foundation for the establishment of high-quality cultivation techniques of S. tonkinens based on microbial community regulation.
      Method High-throughput sequencing was employed to analyze the endophytic bacterial and fungal community structures in 2-year-old (R2), 4-year-old (R4), and 6-year-old (R6) S. tonkinens. This was combined with the determination of alkaloid component content using high-performance liquid chromatography (HPLC). Spearman correlation analysis was then used to elucidate the relationships between the microbial communities and the bioactive components.
      Result With the increase of planting years, the richness and diversity of endophytic bacteria increased significantly, while the diversity and richness of endophytic fungi changed little. The dominant phylum of endophytic bacteria was Proteobacteria, and the dominant phylum of fungi was Ascomycota. The dominant bacterial genera varied with years: R2 and R4 were unclassified_f__Alcaligenaceae, with relatively high abundance of 89.35% and 60.09%, respectively. R6 was Pseudomonas, and the relative abundance was 52.20%. The dominant fungal genera also changed significantly with years, the dominant genus of R2 was Minimelanolocus, the dominant genus of R4 was Codinaea, and the dominant genus of R6 was unclassified_k__Fungi. The content analysis of effective components showed that the content of matrine in R4 was 0.30 mg/g, which was significantly higher than R6 (P< 0.01). The content of oxymatrine was 16.26 mg/g, which was significantly higher than that of R2 and R6 (P< 0.001). The content of cytisine and N-methylcytisine in R6 was 0.99 mg/g and 0.052 mg/g, respectively, both significantly higher than that in R4 (P< 0.01). Correlation analysis showed that endophytic bacteria such as unclassified_f__Alcaligenaceae, Pseudomonas and Pantoea were significantly correlated with active ingredients (P< 0.05). Endophytic fungi of Dactylonectria also had a significant effect on the accumulation of active ingredients (P< 0.05).
      Conclusion This study clarified the potential endophytic bacteria and fungi closely related to the active components of S. tonkinens, revealed the key role of specific functional bacteria in regulating the quality formation of medicinal materials, and provided a scientific basis for further analysis of the mechanism of endophytic bacteria-host interaction affecting the quality of medicinal materials and the formulation of high-quality cultivation strategies.