广西植物2017，Vol.37Issue(5)：634-641,8.DOI:10.11931/guihaia.gxzw201604017
广西柳树桑寄生内生真菌的分离鉴定与抗肿瘤活性菌株筛选
Isolation, identification and antitumor activity of endophytic fungi in Taxilli herba from Salix babylonica in Guangxi
摘要
Abstract
Taxilli herba, the branches with leaves of T.chinensis (DC.) Danser, is a traditionally used Chinese medicine.T.herba has been used to cure powerless bones and tendons, cancer, virus infection, rheumatic arthralgia and so on.However, the endophytic fungi of T.herba has never been studied previously.In this article, the diversity of endophytic fungi in T.herba from Salix babylonica and their antitumor activity were researched.Endophytic fungi were isolated and purified from healthy stems, leaves and roots of Taxilli herba from Salix babylonica in Qinzhou and Guangxi, China.They were identified by morphological characterization firstly, and then the DNA of endophytes was extracted and subjected to ITS sequence analysis.The antitumor activity of ethyl acetate extracts from eight endophytic fungi was tested by MTT method using cancer cell A549 and H460.After preliminary separation, 27 strains of endophytes were isolated from Taxilli herba, and they belong to seven orders, nine genera and fifteen species respectively.Two strains, jin2 and jin30, share sequence max identity of ≤97% with available ITS sequences in NCBI database, and they formed a independent clade in Phylogenetic tree of ITS-rDNA sequences of existing fungi.Therefore, jin2 and jin30 maybe identified as novel species.Pestalotiopsis and Diaporthe were the preponderant genera [accounting for colonization frequencies (CF) 25.9% for each], and all the strains of Pestalotiopsis were isolated from roots.The isolating frequency of Neofusicoccum, Phomopsis and Guignardia were fewer, with three strains for each respectively.Other rare isolated fungi were Penicillium, Fusarium, Colletotrichum and Peyronellaea, with one strain for each.Tissue specificity of endophytes was also observed.For example, Penicillium sp.and Fusarium sp.colonized roots exclusively, Diaporthe arecae, Colletotrichum sp.can only colonized stems, and Guignardia sp.and Peyronellaea sp.only colonized leaves.The antitumor activity showed that one strain, which most related to Pestalotiopsis protearum with ITS sequence similarity at 100%, inhibited the growth of A549 and H460.The ethyl acetate extracts from fungus Gen24 inhibited the growth of A549 and H460 cell at concentration of 800 μg·mL-1, with the inhibiting rate to A549 reached 56.92%, and the inhibiting rate to H460 reached 70.11%.Our research showed the diversity of endophytes from Taxilli herba, and the antitumor endophytes and compounds can be further studied.关键词
柳树/桑寄生/内生真菌/拟盘多毛孢菌/抗肿瘤Key words
Salix babylonica/Taxilli Herba/endophytic fungi/Pestalotiopsis/antitumor分类
农业科技引用本文复制引用
龚斌,巫鑫,韦婷,廖日权,苏本伟,宋静静,江国焕,朱开昕..广西柳树桑寄生内生真菌的分离鉴定与抗肿瘤活性菌株筛选[J].广西植物,2017,37(5):634-641,8.基金项目
国家自然科学基金(81173537, 31560727) （81173537, 31560727）
广西自然科学基金(2014GXNSFBA118135) （2014GXNSFBA118135）
钦州市科技开发项目(201322034) [Supported by the National Natural Science Foundation of China (81173537, 31560727) （201322034）
Guangxi Natural Science Foundation (2014GXNSFBA118135) （2014GXNSFBA118135）
Science and Technology Development Program of Qinzhou (201322034)]. （201322034）
