荞麦拒Na+部位拒Na+的细胞学机制研究

Buckwheat (Fagopyrum esculentum Moench) is a Dicotyledonous Polygonaceae Fagopyrum plant, which is an economic crop with high nutritional value and well health care function. It is strong in stress tolerance, especially in drought and salt stress, and it is also a typical Na+ exclusion plant. On the basis of the determining of main Na+ exclusion localization in buckwheat, this project will study the laws of transportion and allocation of Na+ in main Na+ exclusion localization of buckwheat, especially exploring the characteristics and laws of Na+ compartmentalization in main Na+ exclusion localization cells of buckwheat; and through the cloning of SOS1 and NHX genes of salt tolerant buckwheat variety, the construction and heterologous expression of SOS1-GFP and NHX-GFP of fusion gene expression vector, and the subcellular localization of Na+/H+ antiporter, so as further to find out the relationship between Na+/H+ antiport of plasmolemma and tonoplast of main Na+ exclusion localization in buckwheat and Na+ extrusion and Na+ compartmentalization of cell, and to reveal the cytologic mechanism of Na+ exclusion of main Na+ exclusion localization in buckwheat. These work could provide a theoretical basis and research methods for screening and cultivating stronger salt-tolerant buckwheat varieties.

荞麦（Fagopyrum esculentum Moench）是双子叶蓼科荞麦属植物，是一种具有较高营养价值和良好保健作用的经济作物，它耐逆性强，特别表现在耐旱和耐盐方面，同时它又是一种典型的拒Na+植物。本项目是在确定了荞麦主要拒Na+部位的基础上，采用Na+细胞学定性分析与定量测定方法研究荞麦主要拒Na+部位Na+的运输和分配规律，特别是探明Na+在荞麦主要拒Na+部位细胞中区隔化分布的特点和规律；通过耐盐荞麦品种SOS1和NHX基因的克隆、SOS1-GFP和NHX-GFP嵌合基因表达载体的构建与异源表达，以及Na+/H+逆向转运蛋白的亚细胞定位，进一步搞清荞麦主要拒Na+部位细胞质膜和液泡膜Na+/H+逆向转运与细胞排Na+和Na+区隔化的关系，揭示荞麦主要拒Na+部位拒Na+的细胞学机制，为筛选和培育强耐盐荞麦品种提供理论依据和研究方法。

荞麦是双子叶蓼科荞麦属植物，是一种具有较高营养价值和良好保健作用的经济作物，它耐逆性强，特别表现在耐旱和耐盐方面，同时它又是一种典型的拒Na+植物。本项目采用Na+细胞学定性分析与定量测定方法研究了荞麦主要拒Na+部位Na+的运输和分配规律，探明了Na+在荞麦主要拒Na+部位细胞中区隔化分布的特点和规律，确定了液泡Na+区隔化是耐盐荞麦品种拒Na+的主要方式；通过同源克隆方法获得苦荞麦NHX基因，命名为FtNHX1，GenBank注册登录号KY438929；通过转录组测序，找到苦荞麦拒盐相关基因SOS1，通过RACE方法克隆了该基因全长序列，命名为FtSOS1，GenBank注册登录号KY659584。分别对这两个荞麦关键拒盐基因FtNHX1和FtSOS1进行了生物信息学分析及功能验证，发现拒盐基因的表达与拒Na+部位细胞中Na+区隔化分布的特点相一致，其表达量与荞麦耐盐性呈正相关，从而初步揭示了荞麦拒Na+部位拒Na+的细胞学机制，为筛选和培育强耐盐荞麦品种提供了新的理论依据和研究方法。