矮牵牛DUF620蛋白家族基因PhADR1的功能及调控机理解析

项目名称： 矮牵牛DUF620蛋白家族基因PhADR1的功能及调控机理解析

项目编号： No.31501801

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： 阳永学

作者单位： 武汉市农业科学院

项目金额： 21万元

中文摘要： DUF620 蛋白家族是未知功能结构域（DUF）蛋白家族的重要成员，目前，该家族绝大多数基因的功能尚不清楚。我们前期研究发现，拟南芥DUF620 蛋白家族一个功能未知的基因AtADR1（At5g05840）可能与花药开裂相关。克隆获得的矮牵牛AtADR1同源基因PhADR1在花蕾中高量表达。本项目拟构建PhADR1 RNAi和超表达载体，转化矮牵牛，研究其功能；采用RNA原位杂交和启动子活性检测研究PhADR1时空表达模式；通过基因枪转化洋葱表皮细胞研究PhADR1亚细胞定位；采用酵母双杂交筛选PhADR1互作蛋白，并通过双分子荧光互补（BiFC）在矮牵牛中验证蛋白互作。项目的实施，不仅可以明确矮牵牛PhADR1基因的功能及调控机理，将有助于揭示DUF620家族成员在不同物种中的功能与进化，而且为PhADR1在观赏植物品质改良中的合理利用提供科学依据，具有重要的科学意义和实用价值。

中文关键词： 矮牵牛；PhADR1；；；DUF620蛋白家族；功能分析；调控机理

英文摘要： DUF620 protein family is an important member of domains of unknown function (DUF) protein families. At present, function of most genes is not very clear in DUF620 protein family. Our previous study suggested AtADR1 (locus At5g05840) is likely related to anther dehiscence in Arabidopsis. AtADR1 is a member of DUF620 protein family. The sequence of AtADR1 was cloned from petunia. The bioinformatic analysis suggested that PhADR1 is the homologous gene of AtADR1. PhADR1 was shown to be expressed preferentially in buds in petunia. To investigate the biological function and regulatory mechanism of PhADR1, firstly, the RNAi vector and over-expression vector of PhADR1 will be constructed and transformed with petunia to study the function of PhADR1. Secondly, the spatial and temporal expression pattern of PhADR1 will be studied by RNA in situ hybridization and promoter activity assay of PhADR1. Thirdly, subcellular localization of PhADR1 will be performed with gene gun bombarding into onion epidermal cell. Finally, the interaction proteins of PhADR1 will be screened using yeast two hybrid system and protein interactions will be verified in petunia using bimolecular fluorescent complementation (BiFC). The implementation of the project could not only clarify the biological function and regulatory mechanism of PhADR1 in petunia, but also help to illuminate the function and evolution of members of the DUF620 family in different species and provide a scientific basis for the rational application of PhADR1 in quality improvement of ornamental plants. Thus, this project will has important scientific significance in both theory and practice.

英文关键词： Petunia;PhADR1 ;DUF620 protein family;Functional characterization; Regulatory mechanism