葡萄新型N末端截短质膜H+-ATPase活性调控及其在抗碱性盐胁迫中的作用

项目名称： 葡萄新型N末端截短质膜H+-ATPase活性调控及其在抗碱性盐胁迫中的作用

项目编号： No.31501738

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 农业科学

项目作者： 韩宁

作者单位： 齐鲁工业大学

项目金额： 18万元

中文摘要： 葡萄种植受到土壤盐碱化束缚，因此提高葡萄的抗盐碱性对产业发展非常重要。植物抗盐碱胁迫的重要部位为根，其中根质膜H+-ATPase(质子泵)的活性对植物维持盐碱环境中pH稳恒态至关重要。前期研究中，申请人从盐碱抗性较强的克瑞森无核葡萄根cDNA中克隆了质膜质子泵基因 VvPMA1，酵母功能互补已证实了其强功能，随后序列分析和试验发现克隆的VvPMA1可能是尚未报道的剪接变体，将产生具有较高活性的N末端截短的新型质膜质子泵（VvPMA1β），利于葡萄盐碱抗性。为了验证推测，本项目将对VvPMA1β表达、活性及调控进行进一步分析明确碱性盐胁迫下葡萄是否通过选择剪接产生活性和数量不同的VvPMA1剪接变体影响根质膜质子泵总活性；VvPMA1β是否具有组织、抗性品种特异性；VvPMA1β活性如何调节。希望从基因及蛋白水平了解葡萄根质膜质子泵的表达调控方式，提高盐碱胁迫下葡萄根质膜质子泵活性。

中文关键词： 葡萄；碱性盐胁迫抗性；新型质膜H+-ATPase功能和活性调控；；质膜根H+-ATPase总活性；选择性剪接

英文摘要： Cultivation of grape is restricted due to soil alkalized. So it is important for sustainable development of grape to improve alkali resistance of the grape. The regulation of plasma membrane H+-ATPases activity of roots plays a key role to sustain pH homeostasis of plant in alkali stress. In our previous study, a full length cDNA of root main PM-H+-ATPase isoform VvPMA1 were isolated from alkalinity-insensitive Crimson seedless. Yeast function complementation assay showed that the isolated VvPMA1 had powerful function. Surprisingly, the sequence analysis in grape genomic data and further assay showed that the isolated VvPMA1 might be an undiscovered splicing variant and it would generate a novel N terminus shortened PM-H+-ATPase (VvPMA1β) with higher activity, which would be helpful to alkali resistance of the grape. To verify this speculation, the expression, activity and regulation of VvPMA1β will further be studied. Firstly, it will be explicit whether VvPMA1 will produce different amount and activity of VvPMA1 splicing variant influencing the total activity of root PM-H+-ATPase. Secondly, the organic and variety specificity of VvPMA1β expression will be discussed. Thirdly, how to control the activity of VvPMA1β will be ascertained. This study aims to clarify the regulatory mechanism of PM-H+-ATPase activity of grape root in gene and protein expression level.

英文关键词： grape;resistance of alkali stress;function and regulation of a novel PM-H+-ATPase;PM-H+-ATPase total activity in root;alternative splicing