铁皮石斛内生菌的分离及代谢产物活性的初步研究  

Preliminary study on separation of endophytes from dendrobium officinale and activity of metabolites

作　　者：李俊峰[1] 刘文洪[1] 张贝贝[1] 叶志青[1] 

机构地区：[1]浙江中医药大学,杭州310053

出　　处：《中华中医药杂志》2016年第3期970-974,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.31270051);浙江省教育厅重点项目(No.Z201121522);浙江省中医药科技计划项目(No.2009CA003)~~

摘　　要：目的:分离纯化铁皮石斛的内生菌,对其代谢产物进行抑菌活性和抗肿瘤活性的研究。方法:采用组织块分离法分离铁皮石斛的内生菌,通过形态学观察,初步鉴定分离的内生菌。采用对峙法和牛津杯法分别对内生菌及其发酵代谢产物进行抑菌实验;采用MTT法检测内生菌TG2代谢产物的抗肿瘤活性。结果:共分离得到内生菌31株,其中根9株、茎17株、叶5株。内生菌中真菌22株,细菌9株;且细菌中革兰氏阴性菌7株,革兰氏阳性菌2株。所得31株铁皮石斛内生菌,其中有14株对至少1种供试菌有抑菌作用,占总分离菌株的45.16%。8株内生菌代谢产物的乙酸乙酯相对至少1种供试菌有抑菌作用,占总分离菌株的25.81%。MTT实验表明,内生菌TG2代谢产物具有良好的抗肿瘤效果,且呈剂量依懒性。结论:铁皮石斛含有丰富的内生菌资源,部分内生菌具有代谢抑菌和抗肿瘤活性物质的特性,这可为寻找新型抑菌抗肿瘤物质提供资源。Objective： To separate the endophytes from the dendrobium officinale and study the antimicrobial and antitumor activity of its metabolites. Methods： Endophytes were separated from dendrobium officinale by using tissue separation method and identified by using morphological observation. Antimicrobial tests for endophytes and its metabolites were carried out by using confrontation method and Oxford cup method. The antitumor activity of TG2 as the metabolite of endophytes was detected by using MTT assay. Results： A total of 31 strains of endophytes were separated and obtained, including 9 strains from root, 17 strains from stem and 5 strains from leaf. There were 22 strains of fungi and 9 strains of bacteria in endophytes. Among bacteria strains, there were 7 strains of gram-negative bacteria and 2 strains of gram-positive bacteria. Among 31 strains of endophyte isolated from dendrobium officinale, there were 14 strains which accounted for 45.16% of all separated bacterial strains showed the antimicrobial activity against at least one kind of tested microorganisms, and the ethyl acetate in metabolites of 8 strains of endophyte which accounted for 25.81% of all separated bacterial strains showed the antimicrobial activity against at least one kind of tested microorganisms. MTT results showed that TG2 as a metabolite of endophyte showed excellent anti-tumor activity. Conclusions： Dendrobium officinale contained rich resources of endophytes, and some endophytes showed excellent antitumor activity and bacteriostatic activity, which could provide resources for looking for new antimicrobial and antitumor products.

关 键 词：铁皮石斛 内生菌 分离 抑菌活性 抗肿瘤活性 

分 类 号：R915[医药卫生—微生物与生化药学]