软枣猕猴桃受精后子房蛋白表达的2D-DIGE分析

doi:10.3864/j.issn.0578-1752.2012.17.023

摘要/Abstract

摘要： 【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。

关键词: 猕猴桃, 受精, 子房, 蛋白质组

Abstract: 【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 ?ight/Time of ?ight (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.

Key words: Chinese gooseberry, fertilization, ovary, proteome

齐秀娟, 方金豹, 陈锦永, 顾红, 张绍铃. 软枣猕猴桃受精后子房蛋白表达的2D-DIGE分析[J]. 中国农业科学, 2012, 45(17): 3646-3652.

QI Xiu-Juan, FANG Jin-Bao, CHEN Jin-Yong, GU Hong, ZHANG Shao-Ling. Differential Protein Analysis of Chinese Gooseberry Ovary After Fertilization by a 2D-DIGE Approach[J]. Scientia Agricultura Sinica, 2012, 45(17): 3646-3652.

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