白菜花粉发育相关PME及其互作PMEI基因的表达与功能研究

项目名称： 白菜花粉发育相关PME及其互作PMEI基因的表达与功能研究

项目编号： No.31471877

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 农业科学

项目作者： 曹家树

作者单位： 浙江大学

项目金额： 85万元

中文摘要： 本项目以发现的白菜花粉特异24个PME基因和15个PMEI基因为基础，利用酵母双杂交等技术，找出与PME存在互作关系的PMEI；通过TELEN打靶或amiRNA等方法，分析目的PMEI基因的表达变化对相关PME基因的表达及酶活的影响，探明不同类型的PME与PMEI是否存在以及存在怎样的对应关系；进而选取代表性的PME和PMEI获得表达抑制和过表达转基因植株，采用气相色谱-质谱联用技术测定转基因植株花粉壁和花粉管壁中多糖成分的变化，利用JIM5、JIM7抗体标记不同甲酯化程度的果胶，分析目的基因对细胞壁结构的影响，明确其在花粉发育及花粉萌发、花粉管生长等生殖发育过程中的生物学功能和调控机制。这些研究不仅有助于从整体上认识PME及PMEI基因家族在花粉发育中的作用，加深对花粉壁形成及花粉管生长等过程的果胶代谢的理解，也为全面认识花粉发育及其相关过程的分子机理，有效控制蔬菜等农作物育性提供依据。

中文关键词： 白菜；花粉发育；基因功能；PME；PMEI

英文摘要： On the basis of known 24 pectin methylesterase (PME) genes and 15 PME inhibitor (PMEI) genes specifically expressed in the pollen of Chinese cabbage (Brassica campestris ssp. chinensis), we plan to perform yeast two hybridization and related experiments to find out the PMEI that interacted with PME. By using TELEN targeting or amiRNA methods, we suppose to investigate the changes of PME expression and activity after PMEI gene silencing or over-expressing, and thus to determine whether different types of PME interact with different PMEIs and how they interact with each other. Then select representative PME and PMEI and get gene suppressed and over-expressed transgenic plants. By testing the changes in structure of cell wall polysaccharides of pollen and pollen tube via chromatography-mass spectrometry technology, and methyl esterification degree changes of pectin via JIM5 and JIM7 antibodies labeling, we presume to analyze the influence of selected genes on the cell wall structure and identify the genes' function and their regulation relationship in the process of pollen development, pollen germination and pollen tube growth. These studies will not only deepen our comprehension of the role of PME and PMEI gene family in pollen development and pectin metabolism in the process of pollen cell wall formation and pollen tube growth, but also provide us powerful theoretical basis of the molecular mechanism of pollen development and its related process and effective control measures of the sterility of vegetable and other crops.

英文关键词： Brassica campestris ssp. chinensis;Pollen developmen;Gene function;PME;PMEI