濒危桃儿七根中产鬼臼毒素内生真菌的筛选及分子鉴定  

Screening and molecular identification of podophyllotof xin-producing endophytic ungi from root of endangered Sinopodophyllumhexandrum

作　　者：王婷[1] 马越兴[1] 叶耀辉[1] 郑红梅[1] 张博文[1] 张恩慧[1] 

机构地区：[1]江西中医药大学,江西南昌330004

出　　处：《江西农业大学学报》2017年第2期402-408,共7页Acta Agriculturae Universitatis Jiangxiensis

基　　金：江西省自然科学基金项目(20132BAB205080);江西省专利研发引导与产业化示范专项(省自然科学基金计划)(20143BBM26057)~~

摘　　要：濒危藏药桃儿七根及根茎中含有鬼臼毒素,筛选和鉴定桃儿七根部内生真菌,以期获得产鬼臼毒素类成分的内生菌种,为缓解该药用资源匮乏现状,保护桃儿七植物资源和发现新的天然活性菌株提供参考。采用组织块法对桃儿七根部的内生真菌进行分离,并利用TLC和HPLC检测其代谢产物中的鬼臼毒素类成分,检测波长254 nm,流动相乙腈-水(40∶60)。同时通过形态学和分子生物学相结合的方法对产鬼臼毒素类成分的内生真菌进行鉴定。结果共分离得到8株内生真菌,其中菌株G-1,G-2,G-3,G-4,G-5的发酵液中含有鬼臼毒素类成分,分别鉴定为Pleosporales sp.MF565(100%)、Pseudallescheria sp.T55(100%)、Hypocreales sp.MF489(100%)、Hypocreales sp.LF213(100%)、Cyphellophorareptans strain CBS 458.92(100%)。The root and rhizome of endangered Tibetan SinopodophyUumhexandrum contains podophyllotoxin, screening and identification of endophytic fungi from rootof Sinopodophyllumhexandrum in were carried out to obtain endophytic strains producing podophyllotoxin compounds and provide the reference to alleviate the shortage of the medicinal resources and protect plant resources and discover new natural activitystrains.The endophytic fungi from root of Sinopodophyllumhexandrum were isolated by using the tissue method, the metabolites of podophyllotoxin compounds were detected by thin layer chromatography and high performance liquid chromatography, the detection wave length was 254 nm, mobile phase was acetonitrile-water （ 40： 60）. Combined with morphological and molecular biology methods the podophyllotoxin compounds, which produced endophytic fungi were identified.8 strains of endophytic fungi were isolated, in which G-1, G-2, G-3, G-4, G-5 strains in the fermentation broth contained podophyllotoxin compounds.They were identified as Pleosporales sp.MF565 （ 100% ）, Pseudallescheria sp. T55 （ 100% ）, Hypocreales sp. MF489 （ 100% ）, Hypocreales sp. LF213 （100%） and Cyphellophorareptans strain CBS 458.92 （ 100% ）.

关 键 词：桃儿七 内生真菌 鬼臼毒素 分子鉴定 

分 类 号：R281[医药卫生—中药学]