水稻线粒体基因编码蛋白质在种子萌发及叶片发育过程中的表达分析  

Expression profiling of mitochondrion-encoded proteins in rice seed germination and leaf development

作　　者：兰金苹[1,2] 丁国涛 刘钊[1] 郭亚璐[1] 郭美岑[1] 史佳楠[1] 刘国振[1] 

机构地区：[1]河北农业大学生命科学学院,河北保定071000 [2]河北北方学院生命科学研究中心,河北张家口075000

出　　处：《河北农业大学学报》2016年第2期1-8,共8页Journal of Hebei Agricultural University

基　　金：高等学校博士学科点专项科研基金(20131302110006)

摘　　要：线粒体是真核生物中主要的能量供应细胞器,虽然多种植物的线粒体基因组序列已知,但对线粒体基因编码蛋白质的表达研究关注甚少。本研究采取基于抗体的蛋白质组学策略,制备了17个线粒体基因编码蛋白质的特异性抗体,包括NADH脱氢酶、细胞色素C氧化酶、细胞色素C合成酶、ATP合酶和核糖体蛋白质等5类,利用免疫印迹技术(Western blot)调查了这些蛋白质在水稻种子萌发及不同生长发育时期叶片中的表达情况。结果发现,在种子萌发阶段,ATP合酶中的亚基9和核糖体蛋白质S13亚基表达上调,细胞色素C合成酶CCMB和CCMC表达下调,其余13个表达没有发生变化。在叶片发育阶段,15个蛋白质发生表达上调,且表达模式基本相似,均呈现前期较低,随发育逐渐提高,到成株期达到峰值,其中部分蛋白质在成熟期仍维持在较高的峰值强度(如NAD4、CCMFc、ATP9、RPS1、RPS4和RPS7),提示这些蛋白质在成熟后的灌浆期仍发挥一定作用,而其余蛋白质在成熟期表达量明显下降,提示其功能主要是在营养生长阶段发挥。有意思的是,在叶片发育阶段,RPS19蛋白质表达下调,ATP1蛋白质的表达稳定,它们的特征和功能值得进一步关注。本试验结果直观且相对定量地揭示了线粒体基因编码蛋白质的表达与种子萌发和叶片生长之间的关联,为深入了解其功能提供了有价值的线索。Mitochondrion is the main energy organelle and the primary site for respiration.At present the whole mitochondrial genome sequences of many plants have been released.However,limited information regarding the expression of mitochondrion-encoded proteins is available.Antibody-based rice proteomics strategy was used in this experiment,and 17 specific an-tibodies,including NADH dehydrogenase,cytochrome C oxidase,cytochrome C synthetase,ATP synthase and ribosomal proteins were selected from mitochondrion-encoded proteins.The expression profiling was investigated in seed germination and leaves at different developmental stages by Western blot.The results indicated that 13 proteins were normally expressed in seed germination,except that ATP9 and RPS13proteins were expressed in higher level,while CCMB and CCMC proteins were expressed in lower level.In rice leaves,the expression patterns of 15 proteins were similar and the expression level increased steadily as the growth of leaves.Until the booting and flowering stages,the expressions reached the highest level,and a number of proteins such as NAD4,CCMFc,ATP9,RPS1,RPS4 and RPS7maintained at the highest level in the mature stage.This result implied that these proteins play certain roles in the filling stage,while in contrast,the expressions of rest of the proteins decreased obviously in the mature stage.It is interesting to note that the expression of RPS19 protein decreased and the expression of ATP1 protein was constant,which provided important clues for further research on the characteristics and functions of these proteins.Taking together,this experiment revealed the correlation between the expression patterns of rice mitochondrion-encoded proteins and the process of seed germination and leaf growth in a visual and relative quantitative way,and provided significant information for investigating the function of mitochondrion-encoded proteins.

关 键 词：水稻 线粒体 蛋白质表达 基于抗体的蛋白质组学 免疫印记 

分 类 号：Q5[生物学—生物化学] Q7