转GsPpcK1/ GsPpcK3基因苜蓿耐碱机理的研究

项目名称： 转GsPpcK1/ GsPpcK3基因苜蓿耐碱机理的研究

项目编号： No.31302022

项目类型： 青年科学基金项目

立项/批准年度： 2014

项目学科： 农业科学

项目作者： 才华

作者单位： 东北农业大学

项目金额： 24万元

中文摘要： PpcKs基因除在光合途径起到重要作用外，在干旱及盐胁迫中也起到一定的调控作用，而其在碱胁迫下的作用尚无报道。前期研究中发现，野生大豆PpcKs基因在碱胁迫下持续上调表达，超量表达GsPpcK1/GsPpcK3基因苜蓿的耐碱性明显提高，但其耐碱性的机理尚不明确。本研究以已获得的耐碱转基因苜蓿为材料，采用生理生化及分子生物学手段，通过耐碱生理指标、光合效率及固氮能力的测定，以明确PpcK蛋白提高苜蓿耐碱性的生理机制为重点，确定耐碱与光合效率或固氮的关系；分离光合型或根瘤型PEPC基因，借助药物抑制剂、生化和细胞学途径，分别在叶（茎）或根中构建GsPpcK-MsPEPC级联信号途径，并分析碱胁迫下，GsPpcK-MsPEPC级联关系对光合效率及固氮的调控作用，阐明PpcK蛋白提高苜蓿耐碱的分子机理。综合耐碱生理机制，揭示PpCK调控苜蓿的耐碱机理，并为苜蓿及豆科牧草提供功能明确的耐碱基因资源。

中文关键词： 转基因苜蓿；GsPpcKs；耐碱机理；根瘤固氮；代谢组

英文摘要： Phosphoenolpyruvate carboxylase kinase (PpcK) is a Ca2+ -insensitiv kinase in response to a range of signals in different plant tissues which plays a key role in the control of plant C/N metabolism. Although some reseaches have shown that PpcK gene response to drought or salinity stress,little is known about whether PpcK has the role in adapting to alkali stress.In our earlier study, through microarray analysis, two alkaline (NaHCO3) up-regulated genes GsPpcKs were identified and subsequently cloned from Glycine soja. We transformed these genes into alfalfa and identified that over-expression of GsPpcK1 or GsPpcK3 could enhance alkaline tolerance.While the alkali-resistant mechanism was not clear. As an important extension of our earlier studies, firstly,we would investigate the physiological mechanism of alkali-resistant alfalfa by（i）physiological adaptability, (ii) photosynthetic efficiency, (iii) the activity of N2-fixation.Secondly,we would identify two functional forms of PEPC genes (Phosphoenolpyruvate carboxylase) from Medicago sativa（named as MsPEPCs）, the photosynthetic PEPC gene in leaves or shoots and the non-photosynthetici isoforms in root. Based on the use of selected pharmaceuticals, biochemical and cellular approaches, the aim of this work was to construct the signal transduction cascade of GsPp

英文关键词： transgenic alfalfa；GsPpcKs；alkali-resistant mechanism；symbiotic rhizobium；metabolomics