华泽兰内生真菌的分离及其抑菌、抗肿瘤活性  

Isolation of endophytic fungi from Eupatorium chinense and their antimicrobial, antitumor activities

作　　者：张建芬[1] 林璟 徐好仪 张梦捷 ZHANG Jian-fen;LIN Jing;XU Hao-yi;ZHANG Meng-jie(College of Biology and Environmental Engineering,Zhejiang Shuren University,Hangzhou 310015,China)

机构地区：[1]浙江树人大学生物与环境工程学院

出　　处：《中成药》2020年第1期133-138,共6页Chinese Traditional Patent Medicine

基　　金：浙江省自然科学基金项目(LY18C010002);国家级大学生创新创业训练计划项目(201811842016)

摘　　要：目的研究华泽兰内生真菌的分离及其抑菌、抗肿瘤活性。方法采用组织分离法分离内生真菌,通过形态学观察和其ITS(5.8S)序列的分子生物学方法鉴定内生真菌,KB纸片法和MTT法分别对内生真菌代谢产物的乙酸乙酯提取物进行抑菌实验和体外抗肿瘤实验,PCR法分析和检测内生真菌PKS和NRPS基因。结果从中分离得到8株内生真菌,分别鉴定为禾谷镰刀菌、间座壳属、链格孢属、层生镰刀菌、葡萄座腔菌属小新壳梭孢、细极链格孢、首都叶点霉。菌株EC102、EC103、EC107、EC108、EC111对金黄色葡萄球菌、大肠杆菌、枯草芽孢杆菌、面包酵母都具有很好的抑菌活性,EC102、EC107、EC108、EC109对人乳腺癌细胞株有较好的抗肿瘤活性。从华泽兰内生菌株中发现4个NRPS基因片段和1个PKS基因片段。结论分离得到的内生真菌具有潜在的抑菌和抗肿瘤活性,且能产生NRPS基因和PKS基因。AIM To study the isolation of endophytic fungi from Eupatorium chinense and their antimicrobial and antitumor activities.METHODS The endophytic fungi were isolated by tissue separation method, and identified by morphological observation and molecular biological methods. The antimicrobial activity from the isolated endophytic fungi was detected using filter paper method. And the antitumor activity was detected by MTT assay. PKS and NRPS genes were analyzed by PCR.RESULTS Eight strains of the endophytic fungi were isolated and identified as Fusarium graminearum, Diaporthe sp., Alternaria sp., Fusarium proliferatum, Neofusicoccum parvum, Alternaria tenuissima, and Phyllosticta capitalensis, respectively. The strains of EC102, EC103, EC107, EC108 and EC111 had stronger bacteriostatic activities on S. aureus, B. subtilis, E. coli, and S. cerevisiae. The EC102, EC107, EC108 and EC109 had stronger antitumor activities to human breast cancer cell. Four NRPS gene fragments and one PKS gene fragment were found from the endogenous strain.CONCLUSION The isolated endophytic fungi are potential antimicrobial and antitumor resource, and can produce NRPS and PKS genes.

关 键 词：华泽兰 内生真菌 抑菌 抗肿瘤 

分 类 号：R284.1[医药卫生—中药学]