为探寻新型生物防治方法，揭示茶树精油(TTO)对果蔬采后病原真菌灰葡萄孢霉（Botrytis Cinerea）的抑菌作用机理，以液体培养后收集的B. Cinerea菌丝体为研究对象，进行2倍MIC（最小抑菌浓度）的TTO处理，采用荧光显微镜（FEM）、扫描电镜（SEM）和透射电镜（TEM）研究了处理对细胞膜通透性、菌体形态和超微结构的影响，以及处理过程中菌丝体内活性氧（ROS）积累及相关抗氧化酶类和物质的变化。结果表明：TTO处理后菌丝荧光强度增强，说明其细胞膜通透性明显上升，细胞膜完整性受到破坏；同时菌丝发生严重的皱缩、干瘪，胞浆消化呈碎渣样；TTO导致了菌丝内过氧化氢(H2O2)的大量积累，诱导提高了超氧化物歧化酶（SOD）、过氧化氢酶（CAT）和抗坏血酸-谷胱甘肽（AsA-GSH）循环的活性，说明TTO是通过提高B. Cinerea细胞膜通透性、破坏菌体结构和诱导ROS爆发来发挥抑菌作用。

To investigate a new biological control method and reveal the anti-microbial mechanism of tea tree oil (TTO) against Botrytis cinerea in postharvest fruits and vegetables, the fungal mycelium grown in liquid medium was collected as test material, and then treated with twice MIC (minimum inhibitory concentration) of TTO. Fluorescence electron microscopy (FEM), scanning electron microscopy (SEM), and transmission electron microscopy (TEM) were used to study the effects of the treatment on the membrane permeability, mycelial morphology, and ultrastructure, respectively. Additionally, the accumulation of reactive oxygen species (ROS), changes in enzyme activity, and contents of the substances in mycelia during TTO treatment were explored. The results indicated that after TTO treatment, the fluorescence intensity of the treated group was higher than that of the control, indicating that TTO can increase the cell membrane permeability by destroying its integrity. Additionally, several hyphae were shriveled and dry; their cytoplasm was disintegrated and contained debris-like material. TTO led to the accumulation of hydrogen peroxide (H2O2) in the mycelium and induced higher superoxide dismutase (SOD) and catalase (CAT) activities, as well as ascorbate-glutathione (AsA-GSH) cycle metabolism, in the treatment group than in the control, suggesting that the anti-bacterial activity of TTO is achieved by increasing cell membrane permeability, destroying cell structures, and inducing high ROS production.

国家自然科学基金（31371860）；浙江省科技厅公益性应用研究项目（2013C32094）；宁波市农业和社会发展攻关项目（2014C50084）；宁波大学“水产”浙江省重中之重开放基金（xkzsc1527）