[關鍵詞]
[摘要]
目的 研究延胡索塊莖內生真菌菌群結構及其與延胡索乙素含量的相關性。方法 通過形態(tài)和分子手段相結合的方法對來自8個產地(浙江磐安、浙江東陽、浙江縉云、陜西三合鎮(zhèn)、陜西董家營、重慶開縣、安徽宣城、河南信陽)的延胡索塊莖,進行內生真菌的分離、鑒定及菌群結構分析。同時采用HPLC法測定各產地延胡索塊莖延胡索乙素含量。通過Excel、SPSS軟件分析它們的相關性,以優(yōu)勢屬和種不同水平建立多元線性回歸方程。結果 共分離得到的1 742株內生真菌劃分為19個分類單元,分屬于3個門5個綱10個目14個科15個屬,其中14個鑒定到種水平,5個鑒定到屬水平;8個產地延胡索塊莖中延胡索乙素含量均高于《中國藥典》2015年版標準;浙江產地延胡索塊莖中內生真菌豐富度(S)、內生真菌多樣性指數(shù)(H')和延胡索乙素含量極顯著或顯著高于其他產地;H'值和延胡索乙素含量呈顯著性正相關,木霉屬內生真菌T83與延胡索乙素含量呈最大正相關(相關系數(shù)為0.793)。結論 延胡索塊莖中存在豐富的內生真菌資源;木霉屬內生真菌T83作為優(yōu)勢菌可能與延胡索乙素的積累有關。
[Key word]
[Abstract]
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.
[中圖分類號]
R282.2
[基金項目]
國家自然科學基金面上項目(81673528);浙江省一流學科A類(中藥學)開放基金重點項目(Ya2017001)