Pan Feng, Hou Kai, Gao Feng, Hu Bo, Chen Que, Wu Wei
Agronomy College, Sichuan Agricultural University, Wenjiang 611130, Chengdu, Sichuan, PR China.
Agronomy College, Sichuan Agricultural University, Wenjiang 611130, Chengdu, Sichuan, PR China.
Phytomedicine. 2014 Jul-Aug;21(8-9):1104-9. doi: 10.1016/j.phymed.2014.04.010. Epub 2014 May 20.
Steroidal alkaloids, as the major biologically active components in Bulbus Fritillariae, possess a variety of toxicological and pharmacological effects on humans. The objective of this work was to determine whether endophytic fungi isolated from fresh bulbs of Fritillaria unibracteata var. wabensis can produce one or more alkaloids like its host plant. Four classical reagents including Wagner's, iodine-potassium iodide, Mayer's and improved Dragendorff's were used for primary screening. Then thin-layer chromatography (TLC) and high performance liquid chromatography-evaporative light scattering detection (HPLC-ELSD) were employed to identify the fermentation products of the selected strains. The results showed that extract from one stain (WBS007) has positive reactions in process of primary screening. A further TLC scan and HPLC-ELSD showed that strain WBS007 had two components with the same TLC relative front (Rf) value and HPLC retention time (RT) as authentic peimisine and peiminine. In addition, strain WBS007 was identified as Fusarium sp. based on phylogenetic analysis of ITS sequences. Thus, strain WBS007 produced the bioactive ingredient peimisine and peiminine, as does its host plant, and could be used for the production of peimisine and peiminine by fermentation.
甾体生物碱作为川贝母中的主要生物活性成分,对人体具有多种毒理学和药理学作用。本研究的目的是确定从暗紫贝母鲜鳞茎中分离出的内生真菌是否能像其宿主植物一样产生一种或多种生物碱。使用了包括瓦格纳试剂、碘化钾碘试剂、迈耶试剂和改良的碘化铋钾试剂在内的四种经典试剂进行初步筛选。然后采用薄层色谱(TLC)和高效液相色谱 - 蒸发光散射检测(HPLC - ELSD)来鉴定所选菌株的发酵产物。结果表明,一株菌株(WBS007)的提取物在初步筛选过程中呈阳性反应。进一步的TLC扫描和HPLC - ELSD显示,菌株WBS007有两种成分,其TLC相对比移值(Rf)和HPLC保留时间(RT)与正品浙贝母碱和贝母素甲相同。此外,基于ITS序列的系统发育分析,菌株WBS007被鉴定为镰刀菌属。因此,菌株WBS007与其宿主植物一样产生了生物活性成分浙贝母碱和贝母素甲,可用于通过发酵生产浙贝母碱和贝母素甲。