延胡索块茎中内生真菌菌群结构及其与延胡索乙素含量的相关性研究

目的研究延胡索块茎内生真菌菌群结构及其与延胡索乙素含量的相关性。方法通过形态和分子手段相结合的方法对来自8个产地(浙江磐安、浙江东阳、浙江缙云、陕西三合镇、陕西董家营、重庆开县、安徽宣城、河南信阳)的延胡索块茎,进行内生真菌的分离、鉴定及菌群结构分析。同时采用HPLC法测定各产地延胡索块茎延胡索乙素含量。通过Excel、SPSS软件分析它们的相关性,以优势属和种不同水平建立多元线性回归方程。结果共分离得到的1742株内生真菌划分为19个分类单元,分属于3个门5个纲10个目14个科15个属,其中14个鉴定到种水平,5个鉴定到属水平;8个产地延胡索块茎中延胡索乙素含量均高于《中国药典》2015年版标准;浙江产地延胡索块茎中内生真菌丰富度(S)、内生真菌多样性指数(H′)和延胡索乙素含量极显著或显著高于其他产地;H′值和延胡索乙素含量呈显著性正相关,木霉属内生真菌T83与延胡索乙素含量呈最大正相关(相关系数为0.793)。结论延胡索块茎中存在丰富的内生真菌资源;木霉属内生真菌T83作为优势菌可能与延胡索乙素的积累有关。

Diversity and community structure of endophytic fungi in Corydalis yanhusuo tuber and their correlations with tetrahydropalmatine content

Objective To analyze the diversity and community structure of endophytic fungi in Corydali yanhusuo tuber and their correlations with tetrahydropalmatine content. Methods The endophytic fungi in C. yanhusuo tuber from eight localities (Pan''an of Zhejiang Province, Dongyang of Zhejiang Province, Jinyun of Zhejiang Province, Sanhe of Shanxi Province, Dongjiaying of Shanxi Province, Kaixian of Chongqing, Xuancheng of Anhui Province and Xinyang of Henan Province) were isolated and identified according to the morphology and ITS sequences, so as to analyze the community structure of endophytic fungi; HPLC was used to determine tetrahydropalmatine content, and Excel and SPSS were used to analyze their correlations and establish multiple linear regression equations at different levels of dominant genera and species. Results A total of 1 742 endophytic fungi were isolated and divided into 19 taxa (14 were identified to species level and five to genera level), belonging to 3 phyla, 5 classes, 10 orders, 14 families and 15 genera; The tetrahydropalmatine content in C. yanhusuo tuber from eight localities was higher than the standard of Chinese Pharmacopoeia (2015 edition); The richness (S) and diversity index (H'') of endophytic fungi in C. yanhusuo tuber and tetrahydropalmatine content in Zhejiang Province were extremely significant or significantly higher than those in the other localities. The diversity index (H'') was significantly positive correlated with the tetrahydropalmatine content; There was the largest positive correlation between the tetrahydropalmatine content and T83 (Trichoderma sp.) (correlation coefficient:0.793). Conclusion There are abundant endophytic fungi resources in C. yanhusuo; T83 as a dominant endophytic fungus probably related to the accumulation of tetrahydropalmatine.