The Gene Expression Profile Analysis And Preliminary Studies On Genetic Transformation Of OsRboh1

ROS(reactive oxygen species) is a kind of by-products of organism aerobic metabolism. ROS is involved in growth and development as an important signal molecule besides as toxic secondary metabolites in living organisms. Current study certificate that the production of ROS is mainly controlled by Rboh gene. Alought it has been reported that OsRbohl can play a very important role in Seed germination and stress resistance, we have no idea that if OsRbohl is involved in growth and development of flower organs. Significant up-regulation of OsRbohl expression level in ROS contents higher in the madsS-4 mutant, meanwhile the expression trend of OsRbohl gene and the changed trend of ROS level are in accordant trend in rice anther. These results indicate that OsRbohl gene could be involved in ROS production, little is known about the specific role and molecular bases.To explore the function of OsRbohl gene, we contained recombinant vector pBSK-Rboh-in situ of in situ hybridization probe, we can make a accurate analysis of the expression patterns in riceanther with this pBSK-Rboh-in situ. Our work indicate that OsRbohl expresses principally in 8th to 11th stage of tapetum and microspore cells development by paraffin and in situ hybridization. Based on the previous research, The higher ROS content in 8th to 9th, and the tapetum cellscturn on programmed cell death during 8th and 9th. Thus, we concluded that OsRbohl might be involved in PCD of tapetum cells by regulating production of ROS.To further explore the function of OsRbohl gene, We obtained the expression vectors of pCaMV35S::OsRbohl-amiRNA-1,2,3 depending on the online software WMD (Web MicroRNA Designer 3). Meanwhile we obtained the expression vectors of pUbi.::OsRbohl-RNAi with a specific RNAi fragment by the analysis of BLAST alignment. In addition, we obtained the over expression vectors of pCAMBIA1301-Rboh-OE. There are 78 contained the transgene among regenerated plants with a 100.0% success rate by selection with PCR and hygromycin resistance. The next step in this work is to determin ROS of transgene plants by HPLC or spectrophotometrically in anther. These works may provid the clues and basis for further research the function of the OsRbohl gene in rice ROS product and anther development.