蛋白质赖氨酸乙酰化修饰（KAC）是一种普遍存在的蛋白质翻译后修饰方式，并广泛参与多种生命活动调控。近年来，KAC已成为蛋白质修饰领域的研究热点，但KAC调控植物花粉发育的研究仅限于部分模式植物。申请人前期研究发现，红麻细胞质雄性不育突变体P3A和野生型P3B花粉的蛋白乙酰化修饰存在明显差异，表明KAC可能调控了红麻花粉发育。本项目以突变体P3A为材料，应用基于抗体免疫富集、高分辨率质谱鉴定及Label-Free定量蛋白组学方法，系统研究花粉败育过程中动态KAC蛋白谱；筛选差异KAC蛋白，研究KAC在蛋白功能调控的作用；找出关键KAC蛋白质，并采用免疫共沉淀、Western blot、定点突变以及真核表达等手段进行功能分析；利用CRISPR/Cas9技术以及过表达技术研究明确其在红麻花粉发育中的作用，同时为揭示蛋白乙酰化修饰在植物花粉发育过程中的调控机制提供新的知识。

Protein lysine acetylation (KAC) is a dynamic and reversible post-translational modification (PTM) that play imperative biological roles in diverse cell processes. In recent years, KAC has become a hot topic in the field of protein modification, but the study of KAC regulating pollen development is limited to several model plants, especially for proteome-scale investigations. Different KAC state of kenaf CMS mutant P3A and wild type P3B pollen had been characterized in our previous study. To reveal the possible mechanism of KAC in regulating kenaf pollen development, in the present study, we intend to perform a high resolution label-free based acetylome analysis in mutant P3A based on antibody immune enrichment. Systematically investigate of the dynamic acetylation modified protein profile during pollen abortion. Then screening differentially acetylated proteins and their roles in regulating protein function, characterization key acetylated proteins and their acetylation sites, functional analysis of KAC protein by immunoprecipitation, site-directed mutation, Western blot and eukaryotic expression, and further study on the function of key acetylated proteins by CRISPR/Cas9 gene editing and overexpression technology will be conducted. These should be benefit to clarify the role of key KAC proteins in the pollen development of kenaf, and to provide new knowledge for revealing the role of protein acetylation modification in the development of plant pollen.