铝诱导八仙花柠檬酸分泌转运蛋白基因的克隆和功能分析

项目名称： 铝诱导八仙花柠檬酸分泌转运蛋白基因的克隆和功能分析

项目编号： No.31201656

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 植物保护学、园艺学与植物营养学

项目作者： 陈海霞

作者单位： 湖南农业大学

项目金额： 20万元

中文摘要： 据项目组前期研究发现，八仙花在铝胁迫下能吸收土壤中的铝离子，并将其运输至地上部分，以铝-柠檬酸复合物的状态富集在体内，从而消除铝毒；其中蓝色八仙花(Hydrangea macrophylla cv.Coerulea)抗铝能力最强。本课题在此研究基础上，从蓝色八仙花(H.macropylla cv.Coerulea)中克隆柠檬酸分泌转运蛋白基因，采用RACE技术克隆该基因全长cDNA，然后运用分子信息学技术分析该基因的氨基酸序列及理化性质，再进行序列比对预测基因功能；利用Real-Time RT-PCR法和Western Blot技术分析该基因时空表达特征；再构建八仙花柠檬酸分泌转运蛋白基因过量表达载体转化烟草，通过铝胁迫处理转化植株，分析该基因过量表达与植物耐铝特性的关系。本研究将获得八仙花中第一个耐铝基因，成功揭示八仙花富集铝的分子机理，为重金属污染土壤的治理研究提供新的素材。

中文关键词： 八仙花；铝胁迫；RNA-seq；转录组；转运蛋白

英文摘要： According to previous research, in acidic soil, aluminium ion becomes available to hydrangea roots, stimulating the hydrangea roots to exude citrates. The citrates then form complexes with aluminium ion, these aluminium citrate complexes enter the roots and are transported throughout the shrub, then detoxifying aluminium poison. Among all the cultivars, Hydrangea macrophylla cv.Coerulea has the most ability to resistant aluminium poison. Based on these researches, this project will clone citric acid secretion transport protein gene and its full-length sequence using RACE technique, we focus on analysis of amino acid sequence and physicochemical property using bioinformatics method, then align these sequences and predict its function. Spatio-temporal expression of citric acid secretion transport protein gene will analyzed by Real-Time RT-PCR and Western Blot. Finally, citric acid secretion transport protein overexpression vector will made for the transformation of tobacco, then the relationship of overexpression this gene and aluminum resistance will analyzed by handling suitable aluminum stress. After these studies, we will receive first aluminum resistance genes from hydrangea roots that will reveal the molecular mechanism of accumulating aluminum in hydrangea, and provide new material for heavy metal pollution

英文关键词： Hydrangea；Aluminum stress；RNA-seq；Transcriptome；Transporter