碳酸酐酶BnaβCA1蛋白磷酸化在甘蓝型油菜响应干旱胁迫中的作用机制研究

碳酸酐酶在植物的光合作用中起着重要作用，并受干旱等逆境的诱导。我们前期的研究表明，拟南芥Atβca1突变体对干旱敏感，油菜BnaβCA1蛋白在磷酸化水平上受干旱诱导，但其磷酸化位点的特异性在油菜响应干旱胁迫中的分子机制尚未见报道。本项目将在此基础上，通过磷酸化蛋白组学研究，确定BnaβCA1蛋白中响应干旱胁迫的磷酸化位点；通过创建油菜CRISPR/Cas9敲除株系，研究BnaβCA1基因与干旱的关系；将磷酸化位点突变的BnaβCA1基因分别导入拟南芥和油菜中，研究BnaβCA1磷酸化在干旱中的作用；通过iTRAQ实验，挖掘油菜受BnaβCA1蛋白磷酸化调控的干旱相关蛋白和富集通路。本研究结果将解析BnaβCA1蛋白磷酸化在油菜响应干旱胁迫中的分子调控机制，为培育抗旱油菜新品种提供理论依据。

In higher plants, carbonic anhydrase, which is induced by drought stress, plays an important role in photosynthesis. Our previous research showed that Arabidopsis Atβca1 mutant was more sensitive to drought stress. And its homolog, BnaβCA1 in Brassica napus, was induced by drought stress mainly at the protein phosphorylation level. However, the molecular mechanism of the specificity of its phosphorylated sites in response to drought stress is still unknown. In this project, we will identify the specific phosphorylated sites of BnaβCA1 in response to drought stress by phophoproteomics analyses. The relation of BnaβCA1 and drought stress will be investigated in CRISPR/Cas9 knock-out lines. The function of phosphorylation in response to drought stress will be analyzed by the introduction of BnaβCA1 gene with modified phosphorylation site into Arabidopsis and rapeseed. Finally, iTRAQ analyses will be carried out to verify drought-related proteins and enrichment pathways regulated by phosphorylation of BnaβCA1 in Brassica napus. The results of this project will help us to understand the role of BnaβCA1 phosphorylation, dissect the molecular regulatory mechanism of BnaβCA1 phosphorylation in drought stress process, and provide an important theoretical basis for the genetic breeding of rapeseed with improved resistance to drought stress.