软枣猕猴桃受精后子房蛋白表达的2D-DIGE分析

Differential Protein Analysis of Chinese Gooseberry Ovary After Fertilization by a 2D-DIGE Approach

【目的】了解软枣猕猴桃(Actinidia arguta)受精前和受精后(授粉前和授粉后120 h)子房蛋白质组的变化,为阐明猕猴桃属植物双受精的分子机制提供依据。【方法】应用石蜡切片技术、差异凝胶电泳(DIGE)、质谱(MALDI-TOF/TOF)和生物信息学技术,对其授粉前及授粉后120 h的子房形态及蛋白质组变化进行分析。【结果】授粉后120 h,子房中的胚囊已经完成了双受精;用DeCyder V 6.5软件分析,在授粉前、授粉后120 h的子房中共检测到约1 500个蛋白质点,其中55个有差异表达;质谱鉴定了差异2.0倍以上的蛋白质点13个,其中8个获得理想结果,分别属于6种蛋白质;在6种差异蛋白质中,其中5种在授粉后子房中表达上调,只有1种表达下调。【结论】根据软枣猕猴桃受精前后子房差异蛋白表达谱比较,获得了几种与双受精过程相关的蛋白质分子。

[ Objective ] The changes of Chinese gooseberry （Actinidia arguta） ovary proteins and morphology before and 120 h after pollination were analyzed to provide valuable evidence for further elucidating the molecular mechanism of double fertilization. [Method] Fluorescence paraffin technique,differential in-gel electrophoresis （DIGE）, Matrix-assisted laser-desorption/ionization Time of flight/Time of flight （MALDI-TOF/TOF） and bioinformatic technology were used in the experiment. [Result] The embryo sac in the ovary completed its double fertilization 120 h after pollination. About 1 500 protein points were detected by DeCyder 6.5 （Amersham Bioscience）, and 55 proteins were differentially expressed at ovary before and 120 h after pollination. Thirteen protein spots （differential ratio〉2.0） were identified with MALDI-TOF/TOF, of these proteins,8 belong to 6 kinds of proteins. Five of the six proteins are up-regulated while one is down-regulated. [ Conclusion ] According to the expression differences of the proteins at ovary before and after fertilization, several proteins identified might be related to the process of the double fertilization of Chinese gooseberry.