miR169c/GmNF-YA3模块调控大豆干旱胁迫应答的分子机制研究

干旱影响大豆产量和品质，miRNA与其靶位点组成调控模块为提高大豆抗旱性提供新的有效途径。前期工作发现大豆miR169c/GmNF-YA3调控模块响应干旱并提高拟南芥的抗旱性，但miR169c/GmNF-YA3模块调控大豆干旱胁迫应答的分子机制未知。本项目鉴定转miR169c或GmNF-YA3的大豆毛状根的抗旱性，揭示其在大豆干旱胁迫应答中的功能；利用染色质免疫共沉淀测序和转录组分析寻找GmNF-YA3结合的作用元件和靶基因及相关的代谢及信号转导途径；利用酵母双杂交和双分子荧光互补技术筛选鉴定GmNF-YA3的互作蛋白，探讨GmNF-YA3形成转录调控复合体的组分；选择代表性GmNF-YA3靶基因和互作蛋白，在大豆毛状根中过表达和抑制表达基因鉴定其在大豆干旱胁迫应答中的功能。研究结果将解析miR169c/GmNF-YA3调控大豆干旱胁迫应答的分子网络，为大豆抗旱分子育种提供优异的分子模块。

Drought has severely affected soybean yield and quality. The regulation module of miRNA and its target site provides a new and effective way to improve soybean drought resistance. Previous work found that the soybean miR169c/GmNF-YA3 molecular module plays an important role in drought stress response, but the molecular mechanism of miR169c/GmNF-YA3 module regulating soybean drought stress response is unknown. This project intends to identify the drought resistance of the transgenic hairy roots of miR169c or GmNF-YA3 and reveal its function in response to drought stress in soybean; Identification of cis-acting elements binding by GmNF-YA3, its target genes and related metabolic and signal transduction pathways through chromatin immunoprecipitation sequencing and transcriptome analysis; Screening and identification of GmNF-YA3 interacting proteins using yeast two-hybrid and bimolecular fluorescence complementary technology, and exploring the components of GmNF-YA3 to form a transcriptional regulatory complex; Representative GmNF-YA3 target genes and interacting proteins were selected, and their functions in soybean stress response were identified by overexpressing and suppressing the expression genes in soybean hairy roots. The results will reveal the molecular network of miR169c/GmNF-YA3 regulating soybean drought stress response, and provide an excellent molecular module for molecular breeding of soybean drought resistance.