落叶松干细胞发育关键MicroRNAs功能分析及其TALEN定向突变体创制

项目名称： 落叶松干细胞发育关键MicroRNAs功能分析及其TALEN定向突变体创制

项目编号： No.31330017

项目类型： 重点项目

立项/批准年度： 2014

项目学科： 农业科学

项目作者： 齐力旺

作者单位： 中国林业科学研究院林业研究所

项目金额： 297万元

中文摘要： 围绕林木优良性状形成的生物学基础，在建立了落叶松干细胞发育模式的基础上，针对其特有的PEM发育循环与SE形成时关键的miRNAs表达特性，根据上一项目发现的干细胞发育相关miRNAs，建立关键miRNAs前体的过表达及反义表达体系，增强或抑制这些在关键环节高度表达的miRNAs；引入TALEN技术，进行定向突变体创制。通过落叶松干细胞原胚转化检测体系，鉴定这些miRNAs的生物学功能。在miRNA水平上揭示：（1）干细胞胚性获得相关MiRNAs功能验证及调控机制；（2）PEM（I-III）到SE，ABA（正向）或NPA（抑制）调控原胚发育分子机理；（3）从SE到 mature SE的成熟过程中，畸形胚发生原因及正常胚调控机制。从而在分子水平上综合探索树木生长发育规律，阐释MiRNAs在干细胞品种化繁育中的调控机制，揭示造林树种优良性状形成的生物学基础，具有重要的理论意义。

中文关键词： 落叶松;干细胞发育;MicroRNAs;TALEN;PEM

英文摘要： This project focuses on the formation mechanism of desirable qualities in forestry. To identify the biology mechanism and expression of critical MicroRNAs at Pro-Embryogenic Mass (PEM) development and Somatic Embryo (SE) formation, we will create over-expression and miRNA suppression system to enhance or depress the expression of critical microRNAs which plays a key role during the somatic embryo development, by our somatic embryo development system; create knockout mutant by Transcription Activator-Like Effectors Nuclease (TALEN)-mediated gene targeting method. The biology function of miRNAs will be identified by characterization of transformed-proembryo in larch.The mechanism will be revealed by miRNA techniques in the following fields: (1) Biological function and regulation of critical miRNAs which related with embryogenicity development; (2) Regulation for pre-embryo cell lines development from PEM（I-III）to SE after ABA induction or NPA inhibition. (3) Regulation for abnormal pre-embryo development from SE to mature SE. This project will addresses some fundamental molecular questions in growth and development for tree, mechanism of miRNAs in breeding by embryogenic cell lines and formation of desirable qualities in afforestation species.

英文关键词： Larix spp.;Somatic Embryo Development;MicroRNAs;TALEN;PEM