提高植物体的氮素再循环利用是减少农业中过度的肥料投入而不影响作物产量的有效措施之一，自噬途径作为真核生物中进化保守的对细胞内物质进行周转的过程，在氮素的再循环中发挥了重要作用，其中ATG12和ATG5，ATG8与PE的类泛素化结合途径是自噬过程中的关键步骤，它们对于自噬小泡的延伸和封闭起着重要的作用。我们的前期结果表明超表达自噬体形成标志基因OsATG8b能够提高水稻的氮素利用效率和氮素转移效率。本项目将利用已构建的GFP-OsATG8b转基因水稻和获得的atg12，atg5，atg16突变体为材料，通过检测ATG8-PE的表达和观察GFP-ATG8荧光来分析自噬体的形成，阐明ATG12/ATG5/ATG16复合体在自噬过程中的生化功能，同时通过稳定15N同位素标记等方法，分析氮利用效率和氮转移效率，揭示自噬调控水稻氮循环再利用的分子机制，并为水稻分子育种及耕作中的氮素应用提供策略性指导。

Improve the nutrition use efficiency is recognized as a effective measures to reduce the excessive input of fertilizer and maintain high crop yield in agriculture. Autophagy is an evolutionarily conserved degradation system in eukaryotic cells, and played a important role in nitrogen remobilization. Central to autophagy are two conjugation pathways that attach ATG8 to PE and ATG5 to ATG12, which then help with vesicle prolongation and enclosure. Our study showed the over-expression of OsATG8b can enhance the nitrogen use efficiency and nitrogen remobilization efficiency in rice. We generated GFP-OsATG8b transgenic rice and atg12, atg5, atg16 mutant as research material, it elucidate the biochemical function of ATG12/ATG5/ATG16 protein complex in the autophagic process by detecting the expression of ATG8-PE and observing GFP-ATG8 fluorescence for the formation of autophagosomes; meanwhile, 15N pulse chase analysis can be used to estimate nitrogen use efficiency and nitrogen remobilization efficiency. The Project will reveal the molecular mechanism of autophagy involved in nitrogen remobilization, and provide strategic guidance for nitrogen application in rice molecular breeding and cultivation.