iTRAQ结合2D LC-MS/MS技术鉴定健康和死皮橡胶树胶乳差异表达蛋白  

The Identification of Differentially Expressed Latex Proteins in Healthy and Tapping Panel Dryness( TPD) Hevea brasiliensis Trees by iTRAQ and 2D LC-MS /MS

作　　者：袁坤[1] 王真辉[1] 周雪梅[1] 邹智[1] 杨礼富[1] 

机构地区：[1]中国热带农业科学院橡胶研究所,农业部橡胶树生物学与遗传资源利用重点实验室,海南儋州571737

出　　处：《江西农业大学学报》2014年第3期650-655,共6页Acta Agriculturae Universitatis Jiangxiensis

基　　金：中央级公益性科研院所基本科研业务费专项(YWFZX2008-12);海南省重点科技项目(90107);现代农业产业技术体系建设专项资金(CARS-34-02A)

摘　　要：橡胶树死皮(Tapping panel dryness,TPD)是限制天然橡胶产量提高的关键因子之一,鉴定健康和死皮橡胶树胶乳差异表达蛋白对阐明橡胶树死皮发生机制具有重要意义。研究首次采用同位素标记相对和绝对定量(Isobaric tags for relative and absolute quantitation,iTRAQ)结合二维液相色谱串联质谱(Two-dimensional liquid chromatography-tandem mass spectrometry,2D LC-MS/MS)技术对健康和死皮橡胶树胶乳蛋白质组的差异进行研究。结果显示,有16个蛋白差异表达,其中,在死皮树中表达上调的蛋白有2个,下调的蛋白有14个。进一步应用实时荧光定量RT-PCR对其中6个差异蛋白的基因进行表达分析,发现其表达趋势与蛋白质组水平的研究结果一致。功能分析显示,这些差异蛋白主要与细胞程序性死亡(如TCTP、HSP70、CaM、cysteine protease等)和橡胶生物合成(如SRPP和diphosphomevelonate decarboxylase)相关,从蛋白质组学角度进一步验证了橡胶树死皮发生过程中涉及细胞程序性死亡和橡胶生物合成途径。The tapping panel dryness （ TPD） in Hevea brasiliensis is one of key factors limiting the in-crease of natural rubber production .The identification of differentially expressed latex proteins in the healthy and TPD trees is very important to elucidating the TPD mechanism .In this experiment ,the iTRAQ and 2D LC-MS/MS were first used to study the differences between healthy and TPD tree latex proteomes .The results showed that 16 proteins were expressed differentially , including 2 proteins up-regulated and 14 ones down-regulated in TPD trees .The corresponding genes of 6 differentially expressed proteins were analyzed by real time RT-PCR.It was found that the expression trends of the genes accorded with the results at the proteome level.Functional analysis showed that the differential proteins were mainly involved in programmed cell death （such as TCTP,HSP70,CaM and cysteine protease） and rubber biosynthesis （SRPP and diphosphomevel-onate decarboxylase ） .It was further validated that programmed cell death and rubber biosynthesis pathways were involved in the TPD occurring from the proteomics level .

关 键 词：橡胶树 死皮 ITRAQ 2D LC—MS MS 细胞程序性死亡 橡胶生物合成 

分 类 号：S794.1[农业科学—林木遗传育种]