Streptomyces alboflavus抗真菌物质对串珠镰刀菌的抑制机理研究  

Inhibition Mechanism of Streptomyces alboflavus Antifungal Substance Against Fusarium moniliforme

作　　者：王志芳 李贞景[2] 杨明冠[2] 路来风 王昌禄[2] 李政[3] 张健飞[3] 巩继贤[3] WANG Zhifang;LI Zhenjing;YANG Mingguan;LU Laifeng;WANG Changlu;LI Zheng;ZHANG Jianfei;GONG Jixian(Center of Analytical and Testing, Tianjin Polytechnic University, Tianjin 300387, China;College of Food Engineering and Biological Technology/Key Laboratory of Food Nutrition and Safety, Ministry of Education,Tianjin University of Science and Technology,Tianjin 300457,China;School of Textile Science and Engineering, Tianjin Polytechnic University, Tianjin 300387, China)

机构地区：[1]天津工业大学分析测试中心,天津300387 [2]天津科技大学食品工程与生物技术学院/食品营养与安全教育部重点实验室,天津300457 [3]天津工业大学纺织科学与工程学院,天津300387

出　　处：《食品科学技术学报》2019年第5期64-71,共8页Journal of Food Science and Technology

基　　金：天津市科技计划重点项目(16YFZCNC00700);国家自然科学基金资助项目(31701668);天津市自然科学基金资助项目(17JCQNJC14300)

摘　　要：研究了 Streptomyces alboflavus 产生的抗真菌物质对串珠镰刀菌的抑菌机理。通过pH纸层析和捷克八溶剂纸层析实验对抗菌物质进行定性分析;利用荧光显微镜和扫描电子显微镜观察抗菌物质对串珠镰刀菌细胞膜和细胞壁的破坏作用;通过细胞膜和细胞壁主要组分对抑菌物质的拮抗效果,探索抑菌物质的作用靶点;用高效液相色谱法检测抑菌物质对串珠镰刀菌细胞膜中麦角甾醇的抑制作用。极性实验显示,该物质呈中性,捷克八溶剂纸层析表明,该物质属于非水溶性Ⅰ型抗生素;光学显微镜观察,抑菌物质对串珠镰刀菌孢子萌发有抑制作用,同时使菌丝畸形,顶端、中部出现膨大泡囊,呈“念珠状”;经PI染色后荧光显微镜观察,抗菌物质作用后的菌丝细胞膜通透性改变,进一步通过扫描电子显微镜观察,发现泡囊状菌丝表面凹凸不平,并残留有碎片;抑菌物质对细胞壁和细胞膜的组分拮抗作用和高效液相色谱方法检测显示,抑菌物质影响细胞膜中麦角甾醇的合成。研究结果表明, Streptomyces alboflavus 产生的抗真菌物质发挥抑菌作用时,影响串珠镰刀菌细胞膜中麦角甾醇的合成。本研究可为抑菌物质的鉴定和拮抗菌的应用提供理论参考。The inhibition mechanism of antifungal substance generated by Streptomyces alboflavus against Fusarium moniliforme was investigated.The antifungal substance was qualitatively analyzed by pH paper and Doskochilova system chromatography. The destructive effects of antifungal substance on cell membrane and cell wall of Fusarium moniliforme were observed by fluorescence microscopy and scanning electron microscopy. The antagonistic effects of main components of cell membrane and wall on antifungal substance were explored. Furthermore, the inhibitory effect of antifungal substance on ergosterol in cell membrane of Fusarium moniliforme was detected by HPLC. The antifungal substance was neutral and non-water-soluble type I antibiotics by polarity test and Doskochilova system paper chromatography. Optical microscope was used to observe the inhibition of germination of Fusarium moniliforme spores, the mycelia were distortion, deformation and swelling when treated with the antifungal substance, and the fluorescence microscope revealed that the permeability of mycelial cell membrane was changed. Moreover, the results from scanning electron microscope showed that the surface of vesicular mycelia was uneven and fragments remained. The antagonism of antifungal substance effect on components of cell wall and cell membrane and the detection by HPLC showed that the synthesis of ergosterol in the cell membrane was affected by antifungal substance. The synthesis of ergosterol in the cell membrane of Fusarium moniliforme was affected by the antifungal substance produced by Streptomyces alboflavus . This study laid a theoretical foundation on the identification of antifungal substance and the application of antagonistic bacteria.

关 键 词：白黄链霉菌 串珠镰刀菌 抑菌物质 抑菌活性 抑菌机理 

分 类 号：TS201.3[轻工技术与工程—食品科学]