摘要
为了明确放线菌Hhs.015发酵液中的抑菌活性成分,试验采用硅胶柱层析和凝胶柱层析等分离手段,以苹果树腐烂病菌为靶标菌进行活性跟踪,对发酵液中抑菌活性成分进行提取、分离和纯化,得到2种活性物质。采用ESI-MS、NMR等对活性物质进行结构解析,确定化合物为苯乙酸和染料木素。对苯乙酸和染料木素分别进行抑菌活性测定,结果表明,苯乙酸对苹果树腐烂病菌、小麦全蚀病菌和小麦纹枯病菌具有明显抑制作用,其EC_(50)分别为57.21,166.92和238.23μg/mL,。就对苹果树腐烂病菌菌丝生长的抑制作用而言,苯乙酸(EC_(50)=57.21μg/mL)优于市售药剂70%甲基硫菌灵可湿性粉剂(EC_(50)=154.39μg/mL),因此与阳性对照相比,苯乙酸尚具有一定的开发应用前景。染料木素对玉米大斑病菌和番茄早疫病菌具有抑制作用,其EC_(50)分别为522.40和311.53μg/mL,。利用HPLC方法测定了苯乙酸在发酵过程中的含量变化,发现发酵5 d时发酵液中苯乙酸含量较高。以上结果明确了放线菌Hhs.015发酵液中的部分抑菌活性物质及其抑菌效果,为后续研究提供了参考。
In order to identify the antimicrobial constituent in the fermentation of Hhs. 015, normal phase silica gel, Sephadex LH-20 column chromatography and other separation methods were used to separate the anti-microbial components against the target plant pathogen Valsa mall. Two substances were isolated from the fermentation broth. The exact structures were phenylacetic acid, and genistein, identified through ESI-MS, NMR. Anti-microbial results showed that the EC50 of compound phenylacetic acid against Valsa mali ,Gaeumannomy- ces graminis var tritici and Rhizoctonia cerealis were 57.21, 166.92, 238.23μg/mL. Pheny- lacetic acid showed better antimicrobial activity than 70% Thiophanate-methyl on mycelial growth of Valsa mali, thus phenylacetic acid may have some development and application prospects. The EC50 of genistein against Setosphaeria turcica and Alternaria solani were 522.40,311.53 μg/mL. The exact quantity of phenylacetic acid in fermentation broth was analyzed by HPLC: the content of phenylacetic acid reached the highest at the fifth day. This report clarified parts of the anti-microbial substances and effects from fermentation broth of Hhs. 015, supplied some basis for further study.
出处
《河北农业大学学报》
CAS
CSCD
北大核心
2015年第6期73-78,共6页
Journal of Hebei Agricultural University
基金
国家自然科学基金(31101476
31171796)
陕西省科学技术研究发展计划项目(2013K01-45)
杨凌示范区科技计划项目(2014NY-41)
果树腐烂病防控技术研究与示范(201203034)