摘要： 在高等植物发育过程中,会发生许多形态学和生理学特征上的改变.为了更好地研究草原龙胆特定发育阶段雄蕊内蛋白质组的差异,以草原龙胆栽培品系49号为试材,利用荧光差异双向电泳技术(2D-DIGE)研究了草原龙胆孢原细胞与小孢子母细胞的差异表达蛋白质组,分析了差异表达蛋白质在草原龙胆雄蕊发育过程中的可能作用.结果表明:利用DeCyder软件分析获得了72个主要差异蛋白质点,其中表达上调点为35个,表达下调点为37个.使用MALDI-TOF-TOF质谱仪对胶上消化后的差异蛋白点进行质谱分析,可信度达到90%以上的蛋白质点为15个,分别为5-甲基四氢蝶酰三谷氨酸高半胱氨酸甲基转移酶、S-腺苷甲硫氨酸合成酶、33 kDa光系统Ⅱ放氧复合体、核酮-1,5-二磷酸羧化酶、质体分裂环状蛋白、34 kDa的线粒体外膜膜孔蛋白、70 kDa热激蛋白、磷酸甘油酸激酶、蛋白水解酶复合体β6型亚单位、天冬氨酸蛋白酶、对ATP和Zn²⁺依赖型金属蛋白酶和未知功能蛋白等.因此,在草原龙胆雄蕊发育的特定阶段由于蛋白质组的差异表达,导致雄蕊在形态学和生理学特征上发生改变.

关键词: 荧光差异双向电泳技术(2D-DIGE), 差异蛋白质组, 草原龙胆, 孢原细胞, 小孢子母细胞

Abstract: There are many morphological and physiological changes during ontogeny in higher plants.For the better understanding of the proteomic differences between ontogenetic phases,according to researching the main developmental stages of sporogonium and microsporocyte in cultivars 49 of Eustoma grandiflorum by using fluorescent two-dimensional difference gel electrophoresis (2D-DIGE) carried out proteome differential expression,analysing the functions of the differential proteomics during the different periods of stamens development.The result showed that 72 differentially expressed or modified protein spots were separated by software named DeCyder.While the amount of protein increased in 35 spots during ontogeny and declined in 37 spots.There were fifteen spots whose credibility were as high as 90% had been successfully picked and identified by MALDI-TOF-TOF high-resolution tandem mass spectrometry.Names were 5-methyl 4 hydrogen adjustment acyl glutamic acid homocysteine three methyl transferase,S-adenosine methionine synthetase,33 kDa light system Ⅱ oxygen complexes,1,5-ribulose bishosphate carboxylase,plastid division ring protein,mitochondrial outer membrane protein film hole 34 kDa,heat shock protein 70 kDa,phosphoglycerate kinase,proteolytic enzyme complex beta 6 type subunits,aspartic protease,The ATP and Zn²⁺ dependent metalloproteinases and unknown function protein.Therefore,the differential proteomics expression makes the morphological and physiological changes of the different periods of stamens development.

Key words: 2D-DIGE, Differential proteomics, Eustoma grandiflorum, Sporogonium, Microsporocyte

中图分类号: 

• Q78