拟南芥三个转录因子STF1/2/3介导体细胞调控雌配子体发育的分子机理

项目名称： 拟南芥三个转录因子STF1/2/3介导体细胞调控雌配子体发育的分子机理

项目编号： No.31470284

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 生物科学

项目作者： 秦源

作者单位： 福建农林大学

项目金额： 85万元

中文摘要： 雌配子体提供了双受精的场所，雌配子体正常发育是有性生殖能否顺利进行的前提条件。但在雌配子体发育早期，大孢子母细胞命运如何决定、大孢子母细胞发生数目如何调控、雌配子发生如何起始的分子机理还很不清楚。本项目拟研究的三个转录因子STF1/2/3，是目前唯一所知在紧邻大孢子母细胞的L2层体细胞中特异/优势表达的基因。我们将应用人工microRNA、CREST和CRISPR/Cas9等多种基因沉默技术研究STF1/2/3的生物学功能；并通过酵母双杂交、免疫共沉淀和蛋白质谱技术，以及比较转录组学、瞬时诱导表达和染色质免疫沉淀-测序技术等多种手段筛选STF1/2/3互作蛋白及下游基因，构建STF1/2/3信号调控网络，阐释STF1/2/3介导的体细胞调控雌配子体发育早期事件的分子机理，为解答有性生殖过程中原始生殖细胞命运的决定、配子的发生以及体细胞与生殖细胞互作等关键科学问题，提供重要参考和理论依据。

中文关键词： 大孢子发生；雌配子体发育；转录因子；体细胞与生殖细胞互作；生殖细胞命运决定

英文摘要： Female gametophyte provides a place for double fertilization. Normal development of female gametophyte is the prerequisites of successful sexual reproduction. However, during the early female gametophyte developmental process, the molecular mechanism underlying megaspore mother cell fate decision, control of the number of formed megaspore mother cells and initiation of megagametogenesis are still far from clear. This project intends to reveal the biological functions of three transcription factors STF1/2/3, which are the only gene family known to date specifically or preferentially expressed in the somatic cells adjacent to the megaspore mother cells in the L2 layer. We will apply different gene silencing techniques, such as artificial microRNA, CREST and CRISPR/Cas9 to uncover the roles of STF1/2/3 in female gametophyte development. Yeast two hybridization, Co-IP/MS analysis and comparative transcriptome study, induced transient expression system and chromatin immunoprecipitation - sequencing technologies will be used to screen STF1/2/3 interactive proteins and downstream genes. We aimed to construct STF1/2/3 dependent-signaling networks and reveal the molecular mechanism underlying STF1/2/3 mediated somatic cells control of female gametophyte development. Our finding will shed light on the study of germ cell fate decision, gametes initiation and somatic cell-germ cell interactions.

英文关键词： megasporogenesis;female gametophyte development;transcription factors;somatic cell-germ cell interaction;germ cell fate decision