具有抗氧化活性的北五味子内生真菌的筛选及鉴定-中国药房.pdfVIP

具有抗氧化活性的北五味子内生真菌的筛选及鉴定 ＊ # 赵 玥 ，秦 源，李 娜，李志鹏，范志强，于祥勇，贾景明（沈阳药科大学中药学院，沈阳 110016） 中图分类号 R282.71 文献标志码 A 文章编号 1001-0408（2015）31-4384-05 DOI 10.6039/j.issn. 1001-0408.2015.31.21 摘 要 目的：对具有抗氧化活性的北五味子内生真菌进行筛选及鉴定。方法：利用组织分离法，从野生北五味子的根、茎、叶及 果实中分离内生真菌，分别采用1，1-二苯基-2-三硝基苯肼（DPPH ）自由基清除法及羟自由基清除法筛选具有抗氧化活性的内生 真菌；并对菌株的总DNA 进行提取，利用通用引物ITS 1和ITS4对菌株18S rDNA ITS 序列进行扩增和测序，将测序结果进行同源 性比对分析，确定活性菌株的分类地位。结果：从北五味子中共分离得到23株内生真菌。其中，从根部分离出的编号为GSR-12 的菌株表现出较强的抗氧化活性，其DPPH 自由基清除率和羟自由基清除率分别为87.96％和82.31％；经比对分析后鉴定GSR-12 为粉红粘帚霉（Clonostachys rosea）。结论：分离自野生北五味子根部的1株粉红粘帚霉具有较强的抗氧化活性。 关键词 北五味子；内生真菌；抗氧化活性；鉴定；粉红粘帚霉 Screening and Identification of Endophytic Fungi from Schisandra chinensis with Antioxidant Activity ZHAO Yue ，QIN Yuan ，LI Na ，LI Zhi-peng ，FAN Zhi-qiang ，YU Xiang-yong ，JIA Jing-ming （School of TCM ， Shenyang Pharmaceutical University ，Shenyang 110016，China ） ABSTRACT OBJECTIVE ：To screen and identify endophytic fungi from Schisandra chinensis with antioxidant activity. METH- ODS ：The tissue isolation skill was used to isolate endophytic fungi from roots ，leaves ，stems and fruits of S. chinensis. And anti- oxidant activity of endophytic fungi were screened by DPPH radical scavenging assay and hydroxyl radical scavenging assay. The to- tal DNA were extracted ；the 18S rDNA ITS were amplified and sequenced with primer ITS1 and ITS4；the results of sequencing were analyzed comparatively based on homology to confirm the classification of active strains. RESULTS ：23 strains were isolated from S. chinensis. GSR-12，isolated from roots of S. chinensis ，had strong antioxidant activities. The scavenging rate on DPPH and the hydroxyl radical were 87.96％and 82.31％respectively. GSR- 12 strain was identified as Clonostachys rosea by analyzed com- paratively. CONCLUSIONS ：1strain of C. rosea ，isolated from roots o