GmbHLH25转录因子在丹波黑大豆响应答铝胁迫过程中的作用机理研究

项目名称： GmbHLH25转录因子在丹波黑大豆响应答铝胁迫过程中的作用机理研究

项目编号： No.31260297

项目类型： 地区科学基金项目

立项/批准年度： 2013

项目学科： 农业科学

项目作者： 李昆志

作者单位： 昆明理工大学

项目金额： 56万元

中文摘要： 本项目的前期研究表明丹波黑大豆（简称BS）是一种耐铝能力很强的大豆栽培种，从BS根中分离鉴定的转录因子GmbHLH25在BS受铝胁迫后表达上调，过量表达GmbHLH25增加烟草对铝的抗性。本项目将应用报告基因与该转录因子的融合基因分析它的亚细胞定位和表达模式；通过它与酵母GAL4DNA结合域融合确认它的转录激活作用；应用免疫共沉淀技术分离铝胁迫BS根中与它互作的蛋白；应用DNA芯片技术鉴定它调控的靶基因；用凝胶阻滞法或酵母单杂交技术验证GmbHLH25与靶基因启动子DNA结合特性；分析铝胁迫下BS根中各靶基因的表达谱及靶基因编码蛋白的生理作用；在模式植物转基因体系中进一步验证GmbHLH25及其关键靶基因的功能，阐明GmbHLH25在BS响应铝胁迫过程中的作用机理，研究结果将使我们对大豆应答铝胁迫转录调控的分子机理有更进一步的认识和了解，为改良植物耐铝能力的分子育种提供理论依据和基因资源。

中文关键词： 丹波黑大豆；bHLH30转录因子；铝胁迫应答；转录调控；作用机理

英文摘要： Aluminum (Al) toxicity is a major limiting factor for crop production in acid soils. The key step to efficiently solve this problem is to deeply understand Al tolerance mechanism in plants. The preliminary results of this project showed that Glycine max Tamba (referred to as BS thereafter) is a kind of black soybean cultivars with strong Al tolerance. The transcription of a transcription factor GmbHLH25 gene, which was isolated and identified from BS roots, was upregulated after BS was stressed by Al. Overexpression of GmbHLH25 enhanced the Al resistance of transgenic tobacco. In this study, the subcellular localization and expression pattern of GmbHLH25 protein will be analyzed by the fusion of GmbHLH25 to reporter genes; its transactivation will be verified by the fusion of GmbHLH25 to the DNA-binding domain of GAL4 in a yeast system; the proteins binding with GmbHLH25 will be isolated from BS roots under Al stress conditions by co-immunoprecipitation (CoIP); the target genes regulated by GmbHLH25 will be identified using DNA micrray analysis; the binding of promoter sequences of target genes with GmbHLH25 will be validated by gel retardation analysis or yeast one hybridyation; the expression profiles of target genes in BS roots under Al stress will be analyzed and the pysiological functions of target gene enc

英文关键词： Glycine max Tamba；transcription factor GmbHLH30；Al stress response；transcriptional regulation；functional mechanism