枯茗醛对粉红单端孢(Trichothecium roseum)活性氧和能量代谢的调控机理研究

The essential oil of cumin (Cuminum cyminum) and its predominant component cumaldehyde possess considerable antimicrobial efficacy and their role as antifungal is tightly related to their triggering the augmentation of intracellular reactive oxygen species (ROS) in fungi. This proposed research will conduct the investigation of the variation of ROS metabolism and energy production of conidia after in vitro treatment of Trichothecium roseum with the culmaldehyde. The differential biosynthesis of hyphal mitochondria protein will be analyzed via comparative proteomics techniques. The distribution of ROS in the hyphal body and the modification of hyphal surface figure and subcellular structure will also be explored with the use of confocal laser scanning microscopy, scanning electron microscopy and transmission electron microscopy. All these explorations will elucidate the mechanism of initiation of oxidative stress and regulation of energy metabolism by cumaldehyde in T. roseum hyhpa and unveil the interrelationship between hyphal oxidative stress and energy metabolism under the treatment of culmaldehyde. Finally all these works can theoretically clarify the antifungal mechanism of cumin essential oil.

孜然精油及其主要成分枯茗醛具有良好的抑菌活性，其抑菌作用与促进菌体内部的活性氧（ROS）升高密切相关。本项目采用枯茗醛在体外条件下处理粉红单端孢（Trichothecium roseum），通过生化分析研究处理对菌体孢子活性氧代谢及能量产生的影响，采用比较蛋白组学探讨处理后菌体线粒体蛋白质的差异表达, 借助激光共聚焦、扫描和透射电子显微镜，观察处理后ROS在T. roseum 体内的分布，以及菌体形态和亚细胞结构的变化。阐明枯茗醛引发T. roseum氧化胁迫和调节能量代谢的机理，揭示处理后菌体氧化胁迫和能量代谢的关系，为孜然精油的抑菌机理提供理论依据。

孜然精油对多种果蔬采后真菌具有良好的抑菌效果，其抑菌作用与促进菌体内部的活性氧（ROS）升高密切相关。精油的主要活性成分在其抑菌过程中发挥着重要的作用，本项目用枯茗醛在体外条件下处理粉红单端孢（Trichothecium roseum），通过生化分析研究处理对菌体孢子活性氧代谢及能量产生的影响，采用比较转录组学和比较蛋白组学探讨处理后菌体基因与蛋白质的差异表达, 借助激光共聚焦、流式细胞仪、扫描和透射电子显微镜，观察菌体形态和亚细胞结构的变化。同时研究了枯茗醛处理对菌体致病力的影响。结果表明枯茗醛引发T. roseum氧化胁迫，从转录水平和蛋白表达水平造成多种基因的差异表达，其中包括涉及细胞膜脂类代谢，线粒体膜结构和运输功能，糖代谢基因和能量代谢的基因；处理引起了菌体线粒体膜电位的上升和ATPase及脱氢酶活性的下降；同时处理引起了菌体细胞和亚细胞结构明显的改变，显著地降低了菌体的致病力。枯茗醛引发菌体的氧化胁迫，造成线粒体功能的紊乱，使菌体的能量产生机制遭到破坏，从而使其生长受到抑制。该研究为孜然精油的抑菌机理提供了较为深刻的理论依据。