Key Laboratory of Experimental Marine Biology, Center for Ocean Mega-Science, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China; Laboratory for Marine Drugs and Bioproducts, Pilot National Laboratory for Marine Science and Technology (Qingdao), No. 1 Wenhai Road, Qingdao 266237, China; University of Chinese Academy of Sciences, Beijing 100049, China.
Key Laboratory of Experimental Marine Biology, Center for Ocean Mega-Science, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China; Laboratory for Marine Drugs and Bioproducts, Pilot National Laboratory for Marine Science and Technology (Qingdao), No. 1 Wenhai Road, Qingdao 266237, China.
Int J Biol Macromol. 2020 Jul 15;155:1491-1509. doi: 10.1016/j.ijbiomac.2019.11.127. Epub 2019 Nov 18.
With wide application in agriculture, copper fungicides have undergone three stages of development: inorganic copper, synthetic organic copper, and natural organic copper. Using chitin/chitosan (CS) as a substrate, the natural organic copper fungicide C-coordinated O-carboxymethyl chitosan Cu(II) complex (O-CSLn-Cu) was developed in the laboratory. Taking Phytophthora capsici Leonian as an example, we explored the antifungal mechanism of O-CSLn-Cu by combining tandem mass tag (TMT)-based proteomics with non-targeted liquid chromatography-mass spectrometry (LC-MS)-based metabolomics. A total of 1172 differentially expressed proteins were identified by proteomics analysis. According to the metabolomics analysis, 93 differentially metabolites were identified. Acetyl-CoA-related and membrane localized proteins showed significant differences in the proteomics analysis. Most of the differential expressed metabolites were distributed in the cytoplasm, followed by mitochondria. The integrated analysis revealed that O-CSLn-Cu could induce the "Warburg effect", with increased glycolysis in the cytoplasm and decreased metabolism in the mitochondria. Therefore, P. capsici Leonian had to compensate for ATP loss in the TCA cycle by increasing the glycolysis rate. However, this metabolic shift could not prevent the death of P. capsici Leonian. To verify this hypothesis, a series of biological experiments, such as scanning electron microscopy (SEM), transmission electron microscopy (TEM), and enzyme activity measurements were carried out. The results suggest that O-CSLn-Cu causes mitochondrial injury, which consequently leads to excessive ROS levels and insufficient ATP levels, thereby killing P. capsici Leonian.
铜杀菌剂在农业中应用广泛，经历了无机铜、合成有机铜和天然有机铜三个发展阶段。以壳聚糖（CS）为基质，在实验室中开发了天然有机铜杀菌剂 C 配位 O-羧甲基壳聚糖 Cu(II)配合物（O-CSLn-Cu）。以辣椒疫霉为实例，通过串联质量标签（TMT）-基于蛋白质组学和非靶向液相色谱-质谱（LC-MS）-基于代谢组学相结合的方法，研究了 O-CSLn-Cu 的抑菌机制。蛋白质组学分析共鉴定出 1172 个差异表达蛋白。根据代谢组学分析，共鉴定出 93 种差异代谢物。乙酰辅酶 A 相关和膜定位蛋白在蛋白质组学分析中表现出显著差异。大部分差异表达的代谢物分布在细胞质中，其次是线粒体。综合分析表明，O-CSLn-Cu 可以诱导“Warburg 效应”，增加细胞质中的糖酵解，减少线粒体中的代谢。因此，辣椒疫霉必须通过增加糖酵解率来弥补三羧酸循环中 ATP 的损失。然而，这种代谢转变并不能阻止辣椒疫霉的死亡。为了验证这一假设，进行了一系列生物学实验，如扫描电子显微镜（SEM）、透射电子显微镜（TEM）和酶活性测定。结果表明，O-CSLn-Cu 导致线粒体损伤，进而导致 ROS 水平过高和 ATP 水平不足，从而杀死辣椒疫霉。
