橡胶树大小橡胶粒子的差速离心分离及其差异表达蛋白质的研究  

Studies on the Separation and the Differentially Expressed Proteins of the Large and the Small Rubber Particles of Rubber Trees(Hevea brasiliensis Muell.Arg.)

作　　者：项秋兰[1,2] 代龙军[2] 黎瑜[2] 聂智毅[2] 康桂娟[2] 段翠芳[2] 曾日中[1,2] 

机构地区：[1]海南大学园艺园林学院,海口570228 [2]中国热带农业科学院橡胶研究所/农业部橡胶树生物学重点开放实验室/省部共建国家重点实验室培育基地,海南儋州571737

出　　处：《中国农学通报》2012年第4期18-23,共6页Chinese Agricultural Science Bulletin

基　　金：国家自然科学基金项目"橡胶树响应茉莉酸信号橡胶粒子膜蛋白的鉴定与功能研究"(30860232);国家重点基础研究(973前期)项目"橡胶树橡胶生物合成细胞器:橡胶粒子的蛋白质组学研究"(2007CB116203)

摘　　要：为了鉴定巴西橡胶树胶乳中大小橡胶粒子之间的差异表达蛋白质以及揭示橡胶生物合成的分子机制,采用差速离心的方法,从橡胶树的新鲜胶乳中分离纯化总橡胶粒子、大橡胶粒子和小橡胶粒子,用扫描电镜和SDS-PAGE以及质谱等方法分别检测和鉴定大小橡胶粒子的分离效果及其差异表达蛋白。结果表明,大小橡胶粒子可用差速离心法得到有效的分离和纯化,它们在SDS-PAGE电泳图谱上表现出差异表达的橡胶粒子蛋白,如分子量为22.0kDa、24.5kDa和120kDa蛋白等,并对其中分子量为24.5kDa的小橡胶粒子蛋白(SRPP)进行了质谱分析。橡胶树大小橡胶粒子差速离心法的建立,为大小橡胶粒子差异表达蛋白质的分离和鉴定奠定了一定的基础,将进一步促进橡胶树橡胶粒子的起源和发育,以及橡胶生物合成分子机制的研究。In order to identify the differential expressed proteins between the small rubber particles （SRPs） and the large rubber particles （LRPs） in the latex of Hevea brasiliensis （rubber tree） and to unravel the molecular meehanisms of rubber biosynthesis, SRPs, LRPs and the total rubber particles were completely separated and purified from the fresh latex using differential centrifugation, and further detected through scanning electron microscope （SEM）, respectively. One-dimensional SDS-PAGE analysis demonstrated that the protein profile of the LRPs was different with that of the SRPs, including the differentially expressed proteins with appropriate mass weight of 22.0 kDa, 24.5 kDa and 120 kDa, and so on. One protein expressed only on the SRPs, SRPP, was further identified using nano-LC MS/MS. The separation of LRPs and SRPs from the latex would facilitate both the identification of the differential expressed proteins between LRPs and SRPs, and the further investigation into the molecular mechanisms not only promote the biogenesis and the development, but also promote the rubber biosynthesis of the rubber particle of rubber trees.

关 键 词：橡胶树 橡胶粒子 橡胶粒子蛋白 扫描电镜 SDS-PAGE 质谱分析 

分 类 号：S3[农业科学—农艺学]