杜仲叶和果实中内生真菌的分离及抑菌活性.pdfVIP

中草药 Chinese Traditional and Herbal Drugs 第 47 卷 第 16 期 2016 年 8 月 • 2921 • 杜仲叶和果实中内生真菌的分离及抑菌活性 张维瑞，郭秀春，李 钦，闫兴民，李家梦，温艳华，袁王俊* 河南大学中药研究所，河南 开封 475004 摘 要：目的 从杜仲Eucommia ulmoides 叶和果实中分离具有抑菌作用的内生真菌。方法 采用平板培养法从杜仲叶片和果 实中分离内生真菌，通过对峙法和生长速率法考察这些内生真菌的抑菌活性，并通过 HPLC 法考察其具有代谢产生杜仲有效活 性成分的能力。对具有良好抑菌效果和能够代谢杜仲活性成分的内生真菌进行 ITS 序列测定，鉴定其归属。结果 从健康叶片 和果实中分离到 52 株内生真菌，其中 2 个菌株的提取液中含有绿原酸，11 个菌株至少对 4 种病原菌具有抑制作用，其中 29 号菌株（链格孢属Alternaria sp. ）对芽孢杆菌Fusarium graminearum 的抑制效果最好，其抑制率高达 82.6%。通过与GenBank 数据比对，这些内生真菌分别属于链格孢属Alternaria 、黑孢属Niqrospora 和座囊菌纲（Dothideomycetes ）。结论 从杜仲叶和 果实中分离到 11 株具有抑菌作用的内生真菌，这些具有抑菌作用的内生真菌可用于生物制药和农业产业。 关键词：杜仲；内生真菌；抑菌活性；分子鉴定；芽孢杆菌 中图分类号：R282.15 文献标志码：A 文章编号：0253 - 2670(2016)16 - 2921 - 06 DOI: 10.7501/j.issn.0253-2670.2016.16.025 Antimicrobial activity of endophytic fungi isolated from leaf and fruit of Eucommia ulmoides ZHANG Wei-rui, GUO Xiu-chun, LI Qin, YAN Xing-min, LI Jia-meng, WEN Yan-hua, YUAN Wang-jun Institute of Chinese Materia Medica, Henan University, Kaifeng, 475004, China Abstract: Objective To isolate endophytic fungi from the leaves and fruits of Eucommia ulmoides and evaluate their antimicrobial activity.Methods The endophytic fungi from the leaves and fruits of E . ulmoides were isolated by plate culture method, the antimicrobial activity was evaluated by standoff method, and the ability of producing bioactive compounds of E . ulmoides was investigated by HPLC. Results A total of 52 endophytic fungi were isolated from segments of healthy the leaves and fruits E . ulmoides. Extract of two strains had a retention time identical to that of authentic chlorogenic acid. Eleven out of 52 strains exhibited antimicrobial activity to four or more fungal pathogens. The strain 29 (Alternaria SP.) showed maximum inhibition against Fusarium graminearum, and th