摘要
【目的】探索来自红树老鼠簕AiL3菌株抗菌蛋白对杧果炭疽菌的抑制作用,为微生物抗菌活性物质在果蔬防病保鲜中的利用等提供依据。【方法】以杧果炭疽菌为供试病菌,倍比稀释法测定抗菌蛋白对病菌的最小抑菌与杀菌浓度;电镜扫描观测抗菌蛋白对病菌菌丝生长、孢子萌发的抑制作用并拍照;分别用牛津杯、电导率、考马斯亮蓝、试剂盒及蒽酮比色等方法测定抗菌蛋白处理对病菌胞壁几丁质降解、细胞膜透性、可溶性蛋白含量、菌体DNA及糖类含量变化。【结果】抗菌蛋白对病菌的最小抑菌浓度(MIC)和最小杀菌浓度(MBC)分别为30 mg·L-1和60 mg·L-1;抗菌蛋白处理后,病菌菌丝形态表现为肿胀、膨大直至卷曲坍塌破裂,病菌分生孢子出现皱缩、畸形直至消融;病菌细胞膜通透性和体内可溶性蛋白含量明显增加,病菌糖类代谢物质含量随着处理时间的增长表现为峰形变化,先增加后下降;但抗菌蛋白不能降解病菌细胞壁中的几丁质,对病菌DNA也无明显作用。【结论】初步认为该抗菌蛋白对病菌的抑制可能主要是通过破坏病菌细胞膜、干扰病菌蛋白质及糖类物质代谢等起作用。
[Objective] The inhibition effect of antifungal protein produced by Endophytic Bacterium AiL3 from Mangrove on Colletortrichum gloeosporioides was investigated to provide a theoretical basis for its use in fruit and vegetable disease prevention and storage. [Method] The minimum inhibitory concentration and minimum fungicidal concentration were analyzed by double dilution method after C. gloeosporioides was treated with the AiL3 antifungal protein, and the inhibitory effects on mycelium growth and spore germi- nation were observed by scanning electron microscope as well. The methods of Oxford cup, Conductivity, Coomassie blue, DNA extraction kit, Anthrone colorimetric were used to determined cytoderm chitin degradation, cytomembrane permeability, soluble protein content, pathogen DNA and carbohydrate con- tent respectively. [Results] The results showed that the minimum inhibitory concentration and minimum fungicidal concentration of the protein were 30 mg＂ L-1 and 60 mg＂ L-1, respectively. The mycelium growth and spore germination could be inhibited by treatment with the antifungi protein of AiL3. It was observed that the mycelium collapsed, fractured, shrank, broke and branched and the spores experienced the pro- cess of fullness, shrinkage, deformation up to ablation as the treatment time increased. The structure of cytomembrane could be damaged, which resulted in the increase of permeability and released the cell components and the sugar and protein contents of the mycelium increased first and then decreased after application. But the treatment could not change cytoderm chitin and DNA content of the pathogen. [Con- clusion]The antifungal protein may play a major role in destruction of the cytomembrane and interferingwith mycelial protein and carbohydrate metabolism, which finally causes the disorder of physiological function of pathogen.
出处
《果树学报》
CAS
CSCD
北大核心
2013年第6期1016-1022,共7页
Journal of Fruit Science
基金
广东省科技计划(2012A020602049)
关键词
杧果
红树内生细菌
抗菌蛋白
杧果炭疽菌
抑菌作用
Mango
Marine bacterium
Antibacterial protein
Colletotrichum gloeosporioides
Inhibition effect
