【作者】 郭鸿亮；

【导师】 戴绍军；

【作者基本信息】 哈尔滨师范大学 ， 生物化学与分子生物学， 2011， 硕士

【摘要】 玉簪属(Hosta)植物为百合科多年生草本植物,广泛应用于园林地被、花境和盆栽观赏,是重要的观花观叶的阴生地被植物,也是进行植物叶色变化研究的良好模型。我们运用蛋白质双向电泳技术对玉簪(Hosta plantaginea Aschers)叶色变化过程中形态建成期叶片绿色部分、形态建成期叶片黄色部分、叶色充分显现期叶片绿色部分、叶色充分显现期叶片黄色部分和施肥转绿期叶片的蛋白质样品进行检测分析。利用等电点范围4-7的IPG胶条和12.5%的SDS-PAGE对上述蛋白样品进行了系统的双向电泳分析,在考马斯亮蓝染色的聚丙烯酰胺凝胶上依次得到908±15、917±39、873±30、856±53和839±19个蛋白质斑点。利用ImageMaster 6.0软件分析各个蛋白质斑点相对表达丰度随叶色变化进程的改变,共检测到47个表达丰度变化差异显著的蛋白质斑点,并利用MALDI-TOF-MS/MS技术鉴定了其中33个蛋白质斑点(代表了30种蛋白质)。生物信息学分析表明这些蛋白质隶属于9个功能类群,分别参与蛋白质折叠和转运(28%)、新陈代谢(18%)、光合作用(15%)、蛋白质降解(15%)、能量代谢(9%)、蛋白质合成(6%)、信号转导(3%)、胁迫与防御(3%)和未知功能(3%)。在玉簪(H. plantaginea)叶色变化不同时期表达丰度变化的蛋白质中,参与蛋白质折叠和转运、新陈代谢,以及光合作用的蛋白质表达丰度变化显著。这些结果为深入分析玉簪叶色变化过程中重大生理事件的分子机制提供了依据。同时,在对差异表达蛋白质的分析中,我们发现热激蛋白70、谷氨酰胺合成酶和锌金属蛋白酶具有两个同工型,这些蛋白质主要参与蛋白质折叠和转运、新陈代谢和蛋白质降解。蛋白质同工型在玉簪叶色变化过程中表现出多样性的丰度变化模式对于玉簪叶色变化的研究具有重要意义。更多还原

【Abstract】 Hosta is a perennial and herbaceous plant species in Liliaceae, which is widely applied to gardening and aesthetic enjoyment. Hosta spp. is also a good model to study the mechanism of variation of plant leaves color.In this study, we deployed the technology of two-dimensional polyacrylamide gel electrophoresis to identify the protein samples in the leaves of Hosta plantaginea Aschers during color changed stages (green part of the morphogenesis period, yellow part of the morphogenesis period, green part of the leaves color appearance period, yellow part of the leaves color appearance period and fertilizing recovering green period). By using the pH4-7 IPG gel strips and 12.5% SDS-PAGE gel, the proteins from these distinct stages were separated, respectively. After staining by coomassie brilliant blue, 908±15, 917±39, 873±30, 856±53 and 839±19 protein spots were detected on the gels from each samples. By using ImageMaster 6.0, the differentially-expressed proteins during the process of H. plantaginea leaves color changing were analyzed, 47 protein spots were obviously differentially expressed. By using MALDI-TOF-MS/MS, 33 protein spots (represent 30 proteins) were identified. By using bioinformatics technology, these proteins involved in 9 function categories, shch as protein folding and transporting(28%), metabolism(18%), photosynthesis(15%), protein degradation(15%), energy(9%), protein synthesis(6%), signaling(3%), stress and defense(3%) and unknown function(3%).The expression abundance of proteins involved in protein folding and transporting, metabolism and photosynthesis changed significantly in all the differentially-expressed proteins at different stages we identified. These results are a good evidence for deep analysis the molecular mechanisms of the important physiological event during the process of H. plantaginea leaves color changing. Moreover, the abundances of various isoforms of 3 proteins such as heat shock 70 kDa protein, glutamine synthetase isoform GS1c and zinc metalloprotease were detected in the analysis of differentially-expressed proteins during the process of H. plantaginea leaves color changing, these proteins involved in protein folding and transporting, metabolism and protein degradation. This implied the various expressed pattern of these isoforms played great roles during the process of H. plantaginea leaves color changing.更多还原