陡脉冲电场对水稻纹枯菌的杀灭效果及其机理研究

Rhizoctorzia solani will be selected as the research object, dose-effect curve of irreversible electroporation of steep pulsed electric field (PEF) with different parameters in cell treatment will be researched. Dielectric properties of Rhizoctorzia solani cell under different frequency band steep PEF will be analyzed and electric field model of Rhizoctorzia solani cell will be established to Simulate and calculate the steep PEF parameters of critical electroporation and irreversible electroporation which will be applied to the experiment, while the best threshold dose of critical electroporation and irreversible electroporation of steep PEF in cell treatment will be explored. Mechanism of the Rhizoctorzia solani cells apoptosis caused by steep PEF will be deeply studied, the effect mechanism and change rule of the various physical parameters in Rhizoctorzia solani cell caused by steep PEF will be analyzed through optical microscopy and flow cytometry providing the basises for the further field test. This project will provid solid theory and experimental support for the successful development with completely independent intellectual property rights of a new physical agriculture equipment demonstrating significant academic value and good future in real applying.

以水稻纹枯菌为研究对象，研究不同陡脉冲电场参数处理后细胞不可逆性电击穿的剂量－效应曲线。分析不同频段陡脉冲电场作用下水稻纹枯菌细胞的介电特性，建立水稻纹枯菌的电场物理模型，仿真计算出致水稻纹枯菌细胞临界电穿孔和不可逆性电击穿所需的陡脉冲电场参数并应用于实验，探索陡脉冲电场致水稻纹枯菌细胞发生不可逆性电击穿和临界电穿孔的最佳阈值剂量。深入研究陡脉冲电场杀灭水稻纹枯菌细胞机理，采用光学显微镜和流式细胞仪分析陡脉冲电场对水稻纹枯菌细胞的各种生理参数的影响机制和变化规律，为顺利进入农田试验奠定基础。本项目将为最终成功开发拥有完全自主知识产权的这一新型物理农业设备提供理论和实验依据，具有重要的学术价值和良好的实际应用前景。

水稻纹枯病的无化学残留抑制方法是绿色农业的要求也是最具发展前景的研究方向，本项目选择水稻纹枯菌为处理对象，对陡脉冲电场作用下水稻纹枯菌的抑制效果和机理进行了深入研究。测试了水稻纹枯菌的电特性，建立了丝状真菌的电场处理模型，给出了内外膜跨膜电压的仿真结果以优化处理参数，从电场角度阐释了陡脉冲电场致死水稻纹枯菌的机理。用传统的干重法测试了陡脉冲电场对水稻纹枯菌的抑制效果；通过扫描电镜和透射电镜图片揭示了陡脉冲电场对水稻纹枯菌细胞壁和细胞膜的破坏作用，引发电穿孔导致水稻纹枯菌细胞的死亡；测量了几丁质的含量揭示了对其细胞壁的损伤效应，用激光共聚焦显微镜观察了对微丝骨架的损伤；用细胞膜损伤率和分光光度计法验证了细胞膜的损伤效应；用荧光分光光度计观察了胞外钙离子浓度的变化，明确了陡脉冲电场作用下细胞钙离子浓度失调导致死亡。本项目的完成为实现陡脉冲电场杀灭水稻纹枯菌的农业应用提供了一定的理论依据和实验基础。