IAA关键基因在红松体胚发生中体细胞胚性的获得与保持的作用解析

In previous studies on the transcriptome of endogenous hormones regulation in the Korean pine zygotic embryo development, we found that auxin plays an important role in regulating Korean pine embryo development. In this study, we will treat Korean pine embryonic callus with exogenous auxin, and then monitor the process of Korean pine embryonic callus induction and maintenance with morphological and histological methods to identify spatial and temporal expression characteristics and the role of locus of auxin, and to analyze the specificity of spatio-temporal expression of auxin and embryo related genes. We will explore the relationship between the effects of auxin and acquisition and maintenance of embryonic potential by using the cluster analysis. With combination with the transgenic technology to verify the function of key genes of auxin, we aim to reveal the molecular mechanisms of auxin metabolism and its polar transport in regulating the somatic cells into embryogenic cells and embryogenic maintenance in Korean pine somatic embryogenesis. This study may provide a theoretical basis for revealing the molecular mechanism of embryonic development of Korean pine and other conifer species. It can also provide a reference for the identification of early embryogenic callus and explant selection.

项目申请人在前期红松合子胚发育的转录组数据中初步证实生长素在红松胚胎发育中具有重要作用。本研究拟以红松胚性愈伤诱导保持过程作为研究平台，采用外源生长素处理，通过形态学、组织细胞学监测红松胚性愈伤组织诱导过程，明确生长素时空表达特征及作用位点，分析生长素与胚性相关基因时空表达的特异性，通过聚类分析阐释生长素效应与胚性获得保持的关系，再结合转基因技术对生长素关键基因进行功能验证，揭示生长素代谢及其极性运输调控红松体胚发生中体细胞向胚性细胞转化与胚性维持的分子生物学机制。本研究可为红松及其他针叶树种的胚胎发育分子机理的揭示提供理论基础，为早期胚性愈伤组织的鉴定和外植体的选择提供参考。

本研究以红松胚性愈伤组织诱导保持体系作为研究平台，在优化红松胚性愈伤组织诱导条件的基础上，采用外源生长素处理，通过形态学、组织细胞学监测红松胚性愈伤组织诱导过程，明确内源激素时空表达特征，克隆YUCCAs与SERKs基因，并分析基因时空表达的特异性，阐释生长素效应与胚性获得的关系。本研究可为揭示生长素调控红松体胚发生中体细胞向胚性细胞转化的分子生物学机制提供理论基础，为早期胚性愈伤组织的鉴定和外植体的选择提供参考。