马铃薯块茎发育过程中茉莉酸调控的磷酸化蛋白质组研究

Tuber formation involves many important biological processes, including carbon partitioning,signal transduction and meristem determination.Tuber developmental process also requires the comprehensive regulation of many endogenous hormones,which has become an important reaserch field at present.Investigation of the developmental process of tuber in potato could be better understanding of yield and quality underlying tuber formation and development.In this poject, potato tuber development system in vitro and in vivo were used as reaserch object,and the plant physiology,morphology,bioinformatics,molecular biology,iTRAQ (isobaric tags for relative and absolute quantitation) and other multi-disciplinary methods were applied.The relationship between jasmonic acid (JA) content changes and phosphoproteins in tuber development will be analyzed.These proteins will be function prediction and classification. These new critical proteins and their genes will be cloned and role of these proteins and genes involved in the regulation pathway of JA in tuber development will be examined.The results will clarify the function and regulation mechanism of phosphoproteins involved in JA regulating pathway during tuber development, which could provide a theoretical basis for the formation of potato yield and quality traits and will contribute to help us better understanding of physiological mechanisms underlying potato tuber formation and development.

研究马铃薯块茎发育过程，对揭示地下贮藏器官发育机理，阐明马铃薯产量和品质性状的形成具有重要的理论价值和实践意义。本研究以离体块茎发育体系和活体块茎发育体系的块茎为研究对象，运用表观形态学，植物生理学、蛋白质组学、生物信息学和分子生物学等多学科综合手段，对不同发育阶段块茎的茉莉酸（Jasmonic acid,JA）含量变化、性状特征、糖分积累等进行测定；对过量JA和阻断JA合成后不同发育阶段块茎蛋白进行同位素相对和绝对定量(Isobaric tag for relative and absolute quantitation,iTRAQ)标记，质谱分析，功能预测和分类。解析JA诱导的蛋白质磷酸化调节和控制方式，探究JA调控的磷酸化蛋白质组参与块茎发育的机制及模式，确定JA调控马铃薯块茎发育的途径，分离关键的磷酸化蛋白，并对其功能进行验证。这些研究结果将对揭示马铃薯块茎发育机理提供新思路。

研究马铃薯块茎发育过程中茉莉酸（JA）调控途径和模式，对阐明马铃薯产量和品质性状的形成具有重要理论价值和实践意义。本研究以离体诱导培养体系的块茎为研究对象，对马铃薯块茎发育过程中茉莉酸（JA）和茉莉酸甲酯（MeJA）调控的块茎表观形态、生理生化特征、激素含量、蛋白质组和磷酸化蛋白质组进行分析，获得了块茎发育过程中JA和MeJA调控的关键蛋白质和磷酸化蛋白质信息及其作用机理。取得以下研究结果：1、MeJA促进块茎形成，有利于贮藏物质积累，参与活性氧信号调控块茎发育途径。MeJA诱导的块茎发育相关蛋白质主要通过促进碳源流向糖酵解、三羧酸循环、乙醛酸循环等途径，产生更多能量和促进物质积累。2、JA同样促进块茎形成，加入抑制剂水杨苷羟肟酸（SHAM）后，块茎发育受到抑制，说明JA具有很强的块茎诱导活性。JA诱导的块茎发育相关蛋白质主要涉及生物能量和代谢、储藏、防御、氧化还原平衡、转录和翻译以及转运等功能。通过诱导AGPase、patatin和蛋白酶抑制剂表达来促进块茎中淀粉和储藏蛋白积累，加速碳水化合物代谢为块茎发育提供大量能量，诱导抗氧化酶和病程相关蛋白表达来维持块茎细胞内氧化还原平衡和抗病性。3、对JA诱导的块茎发育磷酸化蛋白质组进行分析，鉴定了172个磷酸化蛋白，包含218个磷酸化位点。这些磷酸化肽被显著富集的基序所代表的蛋白质主要包括：脯氨酸蛋白激酶、分裂原活化蛋白激酶、Ca2+/CaM依赖的蛋白激酶、14-3-3蛋白激酶、细胞周期蛋白依赖性激酶等，它们在块茎发育过程中发挥着重要的磷酸化调控作用。4、JA也促进二倍体马铃薯块茎体积增大和干物质积累，并提前块茎结薯时间。上述这些结果对揭示JA调控马铃薯块茎形成和发育的分子机理具有重要科学意义和应用价值。