一株多花黄精内生真菌的鉴别及其抗菌代谢产物

【目的】药用植物内生真菌是一类重要的微生物资源,能代谢产生多种生物活性物质。本研究从浙江庆元百山祖自然保护区多花黄精(Polygonatum cyrtonema)分离获得1株具有抗菌活性的菌株zjqy610。【方法】通过形态和ITS rDNA序列分析,鉴定为变灰青霉(Penicillium canescens)。采用正相硅胶柱层析和凝胶(Sephadex LH-20)柱层析,以紫外光或碘蒸汽显迹,配合活性追踪等,从zjqy610发酵液中分离获得3个具抗真菌活性的化合物。【结果】通过质谱和核磁共振波谱技术分别将其结构鉴定为:乙基氧苯氨基亚胺乙酸(o-acetylbenzeneamidinocarboxylic acid)、灰黄霉素(griseofulvin)和[1,2-b]呋喃2-甲基3-羧甲基4-羟基-5-甲氧基萘(naphtho[1,2-b]furan-3-carboxylic acid,4-hydroxy-5-methoxy-2-methyl-)。抑菌活性测试表明,3个化合物对多种植物病原真菌具有抑制活性,其中化合物zjqy610D-4对番茄灰葡萄孢、圆形炭疽菌、泻根亚隔孢壳和核盘菌4种病原真菌的活性最强,半抑制浓度EC50分别为0.68、0.38、0.91和0.61mg/L。【结论】该化合物具有开发成农用抗生素的价值。

Isolation and identification of an endophytic fungus of Polygonatum cyrtonema and its antifungal metabolites

Abstract: [Objective] Endophytic fungi in medicinal plants produce a variety of bioactivity compounds. In this study, an endophytic fungus zjqy610 with antifungal activity was isolated from Polygonatum cyrtonema in Zhejiang Qingyuan Baishanzu Mountain nature reserve. [Methods] Strain zjqy610 was identified as Penicillium canescens based on the morphology and its rDNA sequence analysis. Three antifungal compounds were isolated from the fermentation broth of zjqy610 through normal-phase silica gel column chromatography and gel (Sephadex LH-20) column chromatography, traced by ultraviolet light or iodine vapor with bioassay-guided fractionation. [Results] These three compounds were elucidated as o-acetylbenzeneamidinocarboxylic acid (zjqy610B-g-3), griseofulvin (zjqy610D-4) and naphtho [1,2-b] furan-3 -carboxylic acid, 4-hydroxy-5-methoxy-2-methyl-(zjqy610F-2) based on mass spectrometry and nuclear magnetic resonance spectroscopy. The antifungal activity assays showed that the three compounds had inhibitory to variety of plant pathogenic fungi. Compound zjqy610D-4 had strong antifungal activity against Botrytis cinerea, Colletotrichum orbiculare, Didymella bryoniae and Sclerotinia sclerotiorum. EC50 was 0.68, 0.38, 0.91 and 0.61 mg/L, respectively. [Conclusion] Zjqy610D-4 is deserved to develop an agricultural antibiotics.