苹果花芽孕育蛋白质组学初步分析  

Preliminary Proteomics Analysis of the Total Proteins of Flower-Bud Induction of Apple Trees

作　　者：曹尚银[1] 张秋明[2] 朱志勇[1] 郭俊英[1] 陈玉玲[1] 薛华柏[1] 

机构地区：[1]中国农业科学院郑州果树研究所,郑州450009 [2]湖南农业大学园艺学院,长沙410128

出　　处：《中国农业科学》2007年第10期2281-2288,共8页Scientia Agricultura Sinica

基　　金：国家自然科学基金(30671455);国家科研院所社会公益研究专项项目(2005DIA4J047);中国农业科学院院长基金(2004017)

摘　　要：【目的】苹果为中国乃至世界最主要果树之一,但由于童期长、其育种工作一直落后于其它作物。本研究旨在揭示苹果花芽分化机理,缩短苹果童期,加速苹果育种进程。【方法】运用蛋白质组学研究技术(双向电泳技术、生物质谱技术、生物信息学技术)对富士苹果树短枝停长后3～9周的花芽、叶芽进行蛋白质分析研究。【结果】短枝停长后第7周花芽形态分化开始,第7周花芽较叶芽的2-DE图谱有283个蛋白点在表达上有明显的质和量的变化。4个蛋白点(16.4、30.2、40.3、65.1kD)为花芽形态分化开始时初前(花序原始体出现)花芽2-DE图谱所特有;3个蛋白点(39.3、60.2、66.3kD)为初后(侧花出现)花芽2-DE图谱所特有,1个蛋白点(77.1kD)为萼片期花芽2-DE图谱所特有。对富士苹果花芽形态分化开始时的4个特异蛋白点用基质辅助激光解吸P电离飞行时间质谱(MALDI-TOFMS)进行肽质量指纹谱分析,并通过检索不同的数据库进行蛋白质鉴定与功能预测,初步认为第256号(16.4kD)、298号(30.2kD)蛋白点是未知蛋白,第327号(40.3kD)蛋白点是与合成酶有关的蛋白,第367号蛋白点(65.1kD)是一个与转录有关的RNA结合蛋白。【结论】富士苹果花芽形态分化开始时有16.4、30.2、40.3、65.1kD4个特异蛋白、侧花出现时有39.3、60.2、66.3kD3个特异蛋白、萼片出现时有77.1kD1个特异蛋白出现。16.4、30.2kD是未知蛋白,40.3kD是与合成酶有关的蛋白,65.1kD是一个与转录有关的RNA结合蛋白。[ Objective ] Apple is one of the most important fruit trees in the world, due to long juvenile period, apple breeding falls far behind other crops. The aim of this project was to reveal the mechanism of apple flower bud differentiation, short the juvenile period, and expedite apple breeding proceedings. [Method]The specific protein during flower-bud formation in mature spur of apple trees （Malus domestica Borkh.） was studied by employing two-dimensional electrophoresis （2-DE）, bioinformatics, and mass spectrometry （MS）. [Result] The results showeds： compared with leaf-bud, the expression of 283 protein spots in quality and quantity on two 2-DE maps was significantly different at the seventh week flower-bud. Four protein spots （16.4, 30.2, 40.3, 65.1 kD） were observed at flower-bud initial stage 1 （inflorescence appears ） in two-dimensiononal electrophoresis maps; Three protein spots （39.3, 60.2, 66.3 kD） were observed at initial stage 2 （lateral bud appears） in two-dimensiononal electrophoresis maps; One protein spot （77.1 kD） was observed at sepal stage in two-dimensiononal electrophoresis maps. Four specific proteins were observed by specific mass fingerprinting and matrix-assisted laser desorption ionization time of flight mass spectrometry. By analysis of fourspecial proteins spots with peptide mass fingerprinting and matrix-assisted laser desorption ionization time of flight mass spectrometry. When Red Fuji apple begin to spur flower buds. At the same time the identified proteins and their putative functions were presented through the retrieval different database. Spot No.256 （16.4 kD） and 298 （30.2 kD） were unknown proteins and spot No.327 （40.3 kD） was identified as the synthesis enzyme protein. Spot No.367 （40.3 kD） was identified as a RNA-binding protein involved in transcription. [Conclusion] At the beginning of flower bud morphological differentiation in Fuji apple, we detected four specific proteins which were 16.4, 30.2, 40.3, 65.1 kD. Three specifi

关 键 词：苹果 花芽 蛋白质组学 特异蛋白 双向电泳 肽质量指纹谱 

分 类 号：S661.1[农业科学—果树学]