巨尾桉抗寒因子CSD1分子调控网络解析

项目名称： 巨尾桉抗寒因子CSD1分子调控网络解析

项目编号： No.31300497

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： 赵艳玲

作者单位： 华侨大学

项目金额： 23万元

中文摘要： 本项目旨在借助植物双向启动子载体，将与铜锌超氧化物岐化酶1相关的激活因子CCS，miR398的正义和反义基因以两两组合的方式构建并通过叶盘法转化获得转基因巨尾桉。在转基因桉树中研究CSD1的激活途径，通过铜分子伴侣CCS在桉树中的大量表达和基因沉默的两种极端转基因桉树，使用双向蛋白质电泳技术研究CCS表达量降低时桉树的CSD1的另外一条激活途径，分析其中可能存在的调控蛋白。以miR398表达异常的转基因桉树为对象，研究低温胁迫下miR398低表达与CSD1的调控关系，高温状态下miR398超表达与CSD1的调控关系。将已有的7种转基因桉树，通过实时定量PCR技术分析探讨在不同温度条件下转录因子SPL7-miR398-CCS-CSD1的关系。通过以上研究结果绘制CSD1在低温胁迫下的分子调控网络。

中文关键词： 巨尾桉；转基因；CuZnSOD1；CCS；GR

英文摘要： CSD1 is the major superoxide detoxifying enzyme inside cytoplasm catalyzing dismutation of superoxide into oxygen and H2O2. CCS1 is the chaperon, which delivers copper ions to the CSD1 apo-protein.MiR398 binds to the CSD1 and CCS1 transcripts and catalyzes their degradation. However, the relationship between miR398-CSD1-CCS1 under temperature stress is unknown. Cloning of CCS1 and miR398 gene from Eucalyptus and construction of plant expression vector by bidirectional promoter vector, which were sense CSD1 and CCS1, sense CSD1 and anti-sense CCS1, sense CSD1 and miR398, sense CSD1 and anti-sense miR398, transformed to Eucalyptus grandis×E.ophylla by agrobacterium-mediated. The eucalyptus taken sense CSD1 with CCS1 and sense CSD1 with anti-sense CCS1 were used to analysis the two activation pathways of CSD1 identified in many species by two-dimensional gel electrophoresis. The eucalyptus with sense CSD1 and miR398 fusion and sense CSD1 and anti-sense miR398 fusion were used to analyzed whether the accession specific variation depends on miR398 regulation, whether the variation in CSD1 correlates with accession specific regulation of the transcript levels of the other main copper-binding proteins CCS1. SPL7 (for SQUAMOSA promoter binding protein- like7) is essential for this response of miR398.The regulatory mecha

英文关键词： Eucalyptus grandis× E.ophylla；transgenic plant；CuZnSOD1；CCS；GR