龟裂链霉菌M527抗真菌物质的分离鉴定及其在黄瓜枯萎病防治中的应用

doi:10.16409/j.cnki.2095-039x.2016.06.016

中国生物防治学报 ›› 2016, Vol. 32 ›› Issue (6): 783-787.DOI: 10.16409/j.cnki.2095-039x.2016.06.016

龟裂链霉菌M527抗真菌物质的分离鉴定及其在黄瓜枯萎病防治中的应用

路丹丹¹, 赵艳芳¹, 马正¹, 张永勇¹, 王继栋², 俞晓平¹

1. 中国计量大学生命科学学院/浙江省生物计量及检验检疫技术重点实验室, 杭州 310018;
2. 浙江海正药业股份有限公司, 台州 318000

收稿日期:2016-04-16 出版日期:2016-12-08 发布日期:2016-12-07
通讯作者: 俞晓平,博士,研究员,E-mail:yxp@cjlu.edu.cn
作者简介:路丹丹,女,硕士研究生,E-mail:18266848927@163.com
基金资助:
国家自然科学基金（31401792）

Identification of Antifungal Compound from Streptomyces rimosus M527 and Its Application in Biocontrol of Pathogenic Fusarium Wilt on Cucumber

LU Dandan¹, ZHAO Yanfang¹, MA Zheng¹, ZHANG Yongyong¹, WANG Jidong², YU Xiaoping¹

1. Zhejiang Provincial Key Laboratory of Biometrology and Inspection and Quarantine/College of Life Sciences, China Jiliang University, Hangzhou 310018, China;
2. Zhejiang Hisun Pharmaceutical Company Limited, Taizhou 318000, China

Received:2016-04-16 Online:2016-12-08 Published:2016-12-07

摘要/Abstract

摘要： 为探明龟裂链霉菌Streptomyces rimosus M527发酵液中抗真菌成分及其对黄瓜枯萎病菌的拮抗作用，本试验以黄瓜枯萎病菌F. oxysporum f. sp. cucumerinum为活性跟踪指示菌。首先，通过有机溶剂萃取、硅胶柱层析、DiaionHP-20凝胶柱层析、制备型HPLC对活性化合物进行分离纯化，结合MS、1H-NMR和13 C-NMR波谱数据对活性化合物结构进行解析，从菌株M527的发酵液中分离得到活性化合物与龟裂杀菌素一致。结果表明，当龟裂杀菌素浓度为2.20 mg/L时，能够完全抑制黄瓜枯萎病菌的孢子萌发；当龟裂杀菌素浓度为8.78 mg/L时，黄瓜枯萎病菌的菌丝生长受到明显抑制。17.56 mg/L的龟裂杀菌素溶液与传统农药乙蒜素（80%）1000倍稀释液进行黄瓜盆栽试验，施药后14和21 d龟裂杀菌素的治疗效果分别为78.3%和70.6%，乙蒜素的治疗效果为66.7%和42.7%。

关键词: 分离鉴定, 龟裂杀菌素, 黄瓜枯萎病菌, 生物防治

Abstract: Using Fusarium oxysporum f. sp. cucumerinum as an indicator strain, the antifungal active component from the fermentation broth of the strain Streptomyces rimosus M527 were identified and its antagonistic activity against fusarium wilt of cucumber were investigated. After the fermentation broth of S. rimosus M527 was extracted with organic solvent and separated on silica gel column chromatography, the active compound was further purified by chromatography on Sephadex LH-20 and preparative HPLC. Based on the structure analyses of the active compound by MS, ¹H-NMR and ¹³C-NMR spectroscopy, the active compound was identified to be rimocidin. Rimocidin at the concentration of 2.20 mg/L could completely inhibit the spore germination of F. oxysporum f. sp. cucumerinum, while the hyphal growth could be inhibited by 8.78 mg/L rimocidin. Greenhouse experiments on potted cucumber plants demonstrated that therapeutic effect of rimocidin was better than that of ethylicin. After treatment for 14 d and 21 d, the therapeutic effect of rimocidin (17.56 mg/L) against F. oxysporum f. sp. cucumerinum in potted cucumber seeding was 78.3% and 70.6%, respectively, while the 1000 fold diluted ethylicin (80%) was 66.7% and 42.7%, respectively.

Key words: purification and identification, rimocidin, Fusarium oxysporum f. sp. cucumerinum, biological control

中图分类号:

S476

路丹丹, 赵艳芳, 马正, 张永勇, 王继栋, 俞晓平. 龟裂链霉菌M527抗真菌物质的分离鉴定及其在黄瓜枯萎病防治中的应用[J]. 中国生物防治学报, 2016, 32(6): 783-787.

LU Dandan, ZHAO Yanfang, MA Zheng, ZHANG Yongyong, WANG Jidong, YU Xiaoping. Identification of Antifungal Compound from Streptomyces rimosus M527 and Its Application in Biocontrol of Pathogenic Fusarium Wilt on Cucumber[J]. journal1, 2016, 32(6): 783-787.

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龟裂链霉菌M527抗真菌物质的分离鉴定及其在黄瓜枯萎病防治中的应用

Identification of Antifungal Compound from Streptomyces rimosus M527 and Its Application in Biocontrol of Pathogenic Fusarium Wilt on Cucumber

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