The Response Of Pectin Methylesterase Gene To Brassinolide And Nitric Oxide

Pectin 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 BR on PME gene, NO'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.