转运蛋白运输辅助因子PvATF1在蜈蚣草砷高效吸收中的功能和调控研究

砷超富集植物蜈蚣草具有极强的砷吸收、富集能力，被广泛应用于砷污染土壤的植物修复。蜈蚣草中的磷转运蛋白PvPht1具有很强的砷酸盐[As(V)]吸收能力，然而其发挥功能所需的辅助因子及调控机制并不清楚。本项目以蜈蚣草PvPht1的运输辅助因子PvATF1为研究对象，采用原位杂交、GFP融合蛋白等技术手段，明确PvATF1在蜈蚣草中的组织定位和亚细胞定位特征；通过酵母双杂交、半体外Pull-down等互作系统鉴定PvATF1与PvPht1蛋白的结合特征；构建蜈蚣草配子体沉默材料，研究PvATF1在PvPht1靶向定位过程中的作用及在蜈蚣草As(V)吸收中的功能；利用酵母单杂交、LUC-REN体系，筛选、鉴定PvATF1上游转录因子对其的转录调控。研究结果将阐明PvATF1在蜈蚣草高效吸收As(V)过程中的功能与调控机制，为利用基因工程手段提高砷污染土壤的植物修复效率提供重要的基因资源。

Pteris vittata, the arsenic hyperaccumulator, has a very strong ability to absorb and accumulate the arsenic, which is widely used in phytoremediation of arsenic-contaminated soil. Members of phosphate transporter Pht1 in Pteris vittata have strong As(V) transport ability, while the regulation pathway of As(V) absorption was unclear. In this proposal, focusing on PvATF1 (Arsenate Transport Facilitator 1), we will analyze its tissue-specific expression pattern and subcellular localization, using in situ hybridization and GFP fusion protein; verify the binding characteristics between PvATF1 and PvPht1 via yeast double hybrid system and Pull-down assay; identify its function involved in trafficking process of PvPht1 and AsV uptake by using Pteris vittata gametocyte PvATF1 knock down lines; employ yeast one hybrid assay and LUC-REN system to screen and identify the transcription factor on the upstream of PvATF1. Combing the above studies，the function and regulation mechanism of PvATF1 in efficient AsV uptake by Pteris vittata will be clarified. And this study will provide a new molecular approach to improve the efficiency of phytoremediation of As contaminated soil.