基于铝抗性及敏感大豆种质进行铝抗性相关ABC转运蛋白基因的筛选、鉴定及其分子机制研究

项目名称： 基于铝抗性及敏感大豆种质进行铝抗性相关ABC转运蛋白基因的筛选、鉴定及其分子机制研究

项目编号： No.31271744

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 农业科学

项目作者： 陆桂华

作者单位： 南京大学

项目金额： 75万元

中文摘要： 酸性土壤占全球可耕地面积的30%～40%，铝毒是酸性土壤中限制作物产量的关键因子之一。已有研究表明，一些ABC转运蛋白在植物抗铝胁迫过程中起着关键作用,但其分子机制还不很清楚。我们前期的蛋白质组学研究发现ABC转运蛋白可能参与了大豆的抗铝胁迫；然而大豆中含有超过200个预测的ABC转运蛋白基因，其中参与铝抗性的成员具体数量及其分子机制，至今尚不清楚。本项目拟以铝抗性（BX10）和敏感大豆(BD2)种质为材料，根据大豆数据库中ABC转运蛋白家族基因序列，设计特异性引物进行PCR筛选出可能在大豆抗铝胁迫中起关键作用的ABC基因，并采用转基因（导入铝敏感大豆种质中）、qPCR、CLSM、SIMS、ICP-AES等技术手段，鉴定其在大豆抗铝胁迫中的功能并研究其分子机制。本项目的实施和完成，将为抗铝大豆新种质的培育提供潜在的关键靶标，并为深入理解酸铝土壤中植物的抗铝毒机制提供实验与理论依据。

中文关键词： 大豆；铝胁迫；转录组；微小RNA；高通量测序

英文摘要： Aluminum (Al) is one of the major limiting factors for crop production in acid soil which accounts for 30%-40% of the global arable land. Previous studies suggested that some ATP-Binding Cassette (ABC) transporters play key roles in the process of plant Al-resistance, but their molecular mechanism remains not well understood, Our previous proteomics study revealed that ABC transporter of soybean is involved in response to Al stress, however, there are more than 200 putative ABC transporter genes in soybean genome, but the detailed number and molecular mechanism of Al-resistance related ABC genes among them remain unclear till now. In this project we will apply two different soybean genotypes, Al-resistant and Al-sensitive soybeans, to screen the key ABC transporter genes by using PCR methods with the gene-specific primers of the ABC family genes in the GenBank database of soybean, and clarify the molecular mechanism and function of these genes involved in the soybean Al-resistance by the transgenic method (introducing them into the Al-sensitive soybean genotype for function analysis), qPCR, confocal laser scanning microscopy(CLSM), secondary ion mass spectrometry (SIMS) and inductively coupled plasma atomic emission spectroscopy (ICP-AES) methods. The implementation and completion of this project could excavate

英文关键词： Soybean；Aluminum stress；Transcriptome；microRNA；high-throughput sequencing