兰州大学机构库 >生命科学学院
果胶甲基酯酶对油菜素内酯和一氧化氮的响应
Alternative TitleThe Response of Pectin methylesterase to Brassinolide and Nitric oxide
陈徵尼
Thesis Advisor赵志光
2009-05-23
Degree Grantor兰州大学
Place of Conferral兰州
Degree Name硕士
Keyword果胶甲基酯酶 拟南芥 油菜素内酯 一氧化氮 胁迫 GATEWAY 基因克隆
Abstract果胶甲基酯酶(pectin methylesterase,PME)是一种普遍存在于植物细胞中的果胶酶,主要起内源调控植物细胞壁上以及细胞之间果胶含量的作用。为了研究在拟南芥(Arabidopsis thaliana)中油菜素内酯(Brassinolide BR)和一氧化氮(Nitric oxide NO)对PME基因的表达调控,我们进行了一系列实验,获得以下结论: 1. BR和NO对PME基因的表达调控:研究表明,BR可以促进PME基因的表达,NO可以抑制PME基因的表达,并且NO能够抑制BR对PME基因的调控作用,但BR不能影响NO对PME基因的表达调控。这就说明,PME基因是BR和NO共同作用的位点,BR和NO可以共同调控该基因的表达,BR对PME基因的作用位点在NO对该基因作用位点的上游。由此可见,BR和NO的一些生理功能的调节可能是通过PME基因实现的。 2. 检测胁迫条件对拟南芥PME基因的影响:实验证明,盐胁迫,渗透胁迫,冷胁迫能够促进PME基因的表达,热胁迫能够抑制该基因的表达,并且这些胁迫对PME的调节是通过BR来完成的,BR能够响应盐胁迫、渗透胁迫、温度胁迫等植物的逆境胁迫,PME可能是BR响应这些胁迫的一个重要的作用位点。 3. 利用Gateway技术对拟南芥PME基因启动子和编码区的克隆以及植物表达载体的构建:根据已报道的拟南芥PME基因序列,通过PCR技术从拟南芥中扩增得到PME基因的启动子片段及CDS全长,利用Gateway克隆技术,将启动子构建到启动子分析载体pMDC163::CUS上,获得植物表达载体pMDC-PME::CUS,将CDS区构建到细胞定位分析载体pMDC43-GFP上,获得植物表达载体pMDC-PME-GFP,将其分别转入农杆菌GV3101中,通过农杆菌介导转化拟南芥,获得转基因拟南芥植株,为深入研究PME基因在拟南芥组织水平以及细胞水平上的定位奠定基础。
Other AbstractPectin methyl-esterase(PME) is a catalytic hydrolysising pectin which is widely spreaded in the plant cells. The main function of PME is to regulate the pectin content on plant cell walls and that between cells. To study the response of PME gene to Brassinolide (BR) and Nitric oxide (NO) in Arabidopsis thaliana, I did some research and obtained the following conclusions: 1. The regulation of BR and NO on PME: It has been shown that BR could promote the expression of PME gene, whereas NO would inhibite it in my study. Moreover, NO can inhibite the function of the BR’s regulation on PME gene. On the contrary, BR exerts no impact on the NO’s regulation on PME. This signifies that both BR and NO could function the same locus of PME gene. In other words, BR and NO can regulate the expression of PME gene. Furthermore, comparing with the acting locus of NO on PME gene, BR’s acting locus lies on its upstream. It is infered that BR and NO may play the role of their physiological regulation through PME gene. 2. Detect the effect of various stress conditions on PME gene: My study proves that salt stress, osmotic stress and cold stress can increase the expression of PME gene, whereas hot stress can inhibite its expression. Additionally, it is BR that fullfills these regulations. BR can affect such plants’ adversity stress as salt stress, osmotic stress, temperature stress.Conclusively, PME could be an important acting locus on which BR influences these stresses. 3. Clone the promoter and code region of Arabidopsis PME gene by using Gateway technology and construct plant expression vector: According to reproted PME gene sequences of Arabidopsis, I acquired the promoter fragment and full-length CDS of Arabidopsis PME gene by PCR technology. By using Gateway clone technology, I built the promoter onto the promoter analysis vector pMDC163::CUS and got plant expression vector pMDC-PME::CUS. Similarly, I built CDS onto the cell localization analysis vector pMDC43-GFP and got plant expression vector pMDC-PME-GFP. And then they are transfered into Agrobacterium GV3101 respectively. By transformating them into Arabidopsis induced by Agrobacterium, we could obtain transgenic Arabidopsis plants which lay the foundation for further study of locating PME gene in organizational level and cell level.
URL查看原文
Language中文
Document Type学位论文
Identifierhttps://ir.lzu.edu.cn/handle/262010/221285
Collection生命科学学院
Recommended Citation
GB/T 7714
陈徵尼. 果胶甲基酯酶对油菜素内酯和一氧化氮的响应[D]. 兰州. 兰州大学,2009.
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