一株苦参内生真菌的抑菌特性及活性成分的结构鉴定

doi:10.3864/j.issn.0578-1752.2011.15.007

摘要/Abstract

摘要： 【目的】分离药用植物苦参内生真菌，明确其抑菌特性，确定内生真菌BS001发酵产生抗菌物质的种类及结构。【方法】采用常规的组织分离法对内生真菌进行分离，平板菌块法测定抑菌特性，捷克八层析试验确定其极性，柱层析及冷冻干燥的方法对其分离纯化，并利用TLC生物自显影进行跟踪，采用高效液相色谱法（HPLC）测定其纯度，1H-NMR、13C-NMR以及液相色谱-质谱（LC-MS）等方法对其进行结构鉴定。【结果】从苦参种子中分离出1株内生真菌菌株，编号为BS001。该菌株对辣椒疫霉病菌、番茄炭疽病菌、油菜菌核病菌、苹果斑点落叶病菌等病原菌具有抑制效果，其中对番茄炭疽病菌的抑菌直径达2.734 cm。其发酵液经进一步分离纯化后，用波谱学技术对其抑菌活性成分进行鉴定，结果表明其结构为6,7-(2′E)dibutenyl-5,8-dihydroxy-(Z)-cyclooct- 2-ene-1,4-dione。【结论】苦参中存在1株对多种病原菌具有抑制作用的内生真菌，经鉴定为土曲霉（Aspergillus terreu）。在该菌株发酵液中得到一种新抗菌活性物质，可用于生物药物及生物农药的开发。

关键词: 苦参, 内生真菌, 抑菌物质, 结构鉴定, 分离, 生物自显影

Abstract: 【Objective】The objective of this study is to isolate the endophytic fungi from Sophora flavescens and define its antimicrobial action. Also the identification of the category and structure of the active substance fermented from Sophora flavescens was conducted. 【Method】Tissue isolation and selective culture medium method was used to isolate the endophytic fungi. Plate method was applied to measure the antimicrobial characters. Eight kinds of solvent system was used to define its polar. Column chromatography and freeze drying, TLC biological autoradiography and HPLC were employed to isolate and purify, trace, and measure the purity, respectively. The structure was analyzed using 1H-NMR, 13C-NMR and LC-MS. 【Result】 An endophytic fungus strain was isolated from Sophora flavescens seed, numbered BS001. It inhibited the growth of Phytophthora capsici, Colletotrichum coccodes, Sclerotinia sclerotiorum, and Alternaria mali. The inhibition to Colletotrichum coccodes was the biggest with the antibacterial diameter of 2.734 cm. After isolating and purifying the fermentation liquor of BS001, the structure of bacteriostatic active composition was 6,7-(2′E)dibutenyl-5,8-dihydroxy-(Z)-cyclooct-2-ene-1,4-dione, which identified by spectroscopy. 【Conclusion】An endophytic fungus, BS001, which has inhibition to some pathogens, was got from Sophora flavescens. Also a new antimicrobial substance 6,7-(2′E)dibutenyl-5,8-dihydroxy-(Z)-cyclooct-2-ene-1,4-dione was extracted from fermented broth of BS001. This antmicrobial substance can be used for development of biopharmaceuticals and biopesticides.

Key words: Sophora flavescens, endophytic fungus, antimicrobial substance, structure identification, isolation, biological autoradiography

何璐, 纪明山, 王勇, 徐洪波, 齐补坤. 一株苦参内生真菌的抑菌特性及活性成分的结构鉴定[J]. 中国农业科学, 2011, 44(15): 3127-3133 .

HE Lu, JI Ming-Shan, WANG Yong, XU Hong-Bo, QI Bu-Kun. Antimicrobial Action of an Endophytic Fungus from Sophora flavescens and Structure Identification of Its Active Constituents[J]. Scientia Agricultura Sinica, 2011, 44(15): 3127-3133 .

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