猪苓是珍贵的药用真菌，菌核入药，在我国用药历史悠久。然而目前野生菌核资源逐渐贫乏。本项目在发现内源性草酸在菌丝和菌核的含量存在显著差异且与氧化应激密切相关及不同浓度外源性草酸可促进或抑制菌核形成后，以nano LC-MS/MS乙酰化修饰蛋白质组学技术获得乙酰化修饰蛋白标记物并阐述草酸/氧化应激影响猪苓菌核发育中能量代谢的分子机理；利用免疫胶体金技术、Western blot及MRM定量检测技术研究在猪苓菌核与菌丝间差异表达且与细胞间粘连及细胞骨架组成密切相关的integrin ß1，actin与tubulin的亚细胞结构定位与分布及表达情况，揭示其在菌丝融合、细胞骨架重塑中的作用。应用非损伤微测技术对草酸影响猪苓菌核形成中Ca2+与H2O2进行定量实时监测，揭示Ca2+与活性氧的变化规律及菌核形成中草酸与Ca2+、H2O2变化的关系。本项目将为草酸影响猪苓菌核发育的分子机制奠定基础。

Polyporus umbellatus is one of the most precious medicinal fungi, which has been used as medicine for more than two thousand years. However, wild sclerotia of P. umbellatus are almost in danger of extinction now..This study is based on our previous study which indicates that endogenous oxalic acid content differs very much in P. umbellatus mycelia and sclerotia and it is closely associated to oxidative stress. Furthermore, different concentrations of exogenous oxalic acid affect sclerotial formation at different manners. The study conducts to investigate the differential genes affected by oxalic acid using nano LC-MS/MS acetylated modification proteomics technology, mainly paying attention to oxalic acid and oxidative stress affecting energy metabolism regulation during P. umbellatus sclerotial formation and acquiring protein markers after acetylated modification. In the previous study, integrin ß1, actin and tubulin were found to be significantly and differentially expressed in mycelia and sclerotia during P. umbellatus sclerotial formation, which were closely associated with adhesion between cells and being main components of cytoskeleton. Based on their subcellular localization, distribution and expression characteristics using Immunogold technique, western blot and quantitative MRM detection assay, it will facilitate to discover what roles integrin ß1, actin and tubulin will play during mycelia fusion and cytoskeleton remodeling affected by oxalic acid. Furthermore, Ca2+ and H2O2 are quantitative real-time dynamic monitored using Non-invasive Micro-test Technology during different stages of sclerotial formation affected by oxalic acid, which will help to elucidate the changing rules of Ca2+ and ROS and the relationship among oxalic acid and changes of Ca2+ and ROS. This study will provide us information for molecular mechanism induced by oxalic acid in P. umbellatus sclerotial formation.