Puroindoline蛋白（PIN蛋白）不仅影响小麦籽粒硬度形成，还能有效抑制许多植物病原菌的生长。目前，国内外对PIN蛋白在小麦后熟期的生化特性及其抗霉菌的作用机制还不十分明确。本项目选取新收获小麦在模拟实验仓中储藏，后熟过程中分离纯化PIN蛋白，研究其与淀粉粒结合作用、对脂质体膜结构的影响及抗菌活性；采用PCR技术定点突变PIN蛋白TRD结构域，分析其与脂膜的结合作用；采用电镜观察后熟期小麦PIN蛋白对储粮主要危害性霉菌灰绿曲霉和产毒黄曲霉菌丝与孢子形态的影响；测定PIN蛋白对菌丝生长、细胞壁合成、细胞膜结构稳定性与通透性、线粒体呼吸作用、DNA合成等方面的影响；采用转录组学技术研究PIN蛋白对霉菌营养生长及产毒代谢的调控作用。本项目从PIN蛋白生化特性入手，从形态学、生理学和遗传学方面解析PIN蛋白抗霉菌的作用机制，为耐储藏小麦品种的选育及新型储粮防霉剂开发提供科学依据。

The Puroindoline (PIN) is a critical protein involved in wheat hardness and frequently found abundant in endosperm and aleurone layer of mature seed. Previous results showed that PIN not only regulate the formation of wheat hardness during seed development but also effectively inhibit the growth of many plant pathogens. The after-ripening period is a key stage for wheat hardness formation, however, the role of PIN based on adhesion with the lipid membrane and its impact on membrane structure are not yet clear, the biochemical properties therefore need more investigation. In addition, the fungal resistance of PIN also remain to be elucidate.. In our work, newly harvested wheat with different hardness properties will be selected to mimic after-ripening in a simulated grain storehouse，the PIN protein was subsequently extracted and purified during storage. The quantity of PIN binding with starch granules, effects on structure of artificial liposome membrane and its fungus resistance were studied. By using overlap PCR technology, the important active center of PIN protein TRD domain was carried out site-directed mutation, and therefore the role of PIN in lipid binding effect was evaluated. The influence of PIN on morphology of hyphae and spores of Aspergillus glaucus and Aspergillus flavus, the two main pathogenic fungi frequently involved in cereal spoilage during grain storage, are evaluated with electron microscope. Additionally, the fungal cell wall synthesis, structure and permeability of cell membrane, respiration of mitochondria, synthesis and degradation of nucleic acid of these fungi affected by PIN protein will be investigated. RNA-seq technology is also employed to elucidate the regulation of PIN protein on vegetative growth and metabolism of aflatoxin of Aspergillus flavus. In our work, the resistance mechanism of PIN protein will be studied from morphology, physiology and genetics of the test fungi.. In view of this, our work will not only enrich the evidence for the hypothesis of wheat hardness formation, but also support molecular breeding of endurable storage wheat and potentially develop new preservative used in grain storage.