Expression Analysis Of The Promoter Of ANK And Pita In Transgenic Rice

Gene expression is regulated largely by the interaction of its promoters and specific cis-elements, so the expression pattern and the underlying molecular mechanism analysis is a feasible approach for the functional identification of the candidate resistance gene. In the long process of evolution, plants has developed inducible resistance which limit the defense reaction against pathogens within the area around the site of infection, minimizing the waste of substance and energy and as well as the autotoxicity of toxic secondary metabolites which can kill germs. So, analysis of promoter expression pattern of defense reaction associated genes can not only help to identify the function of the defense reaction associated genes, but perhaps can also offer inducible promoters for the diseases resistance gene engineering to improve the expression of exogenous resistance genes. In this paper, the promoter expression pattern of OsANKp and OsPITAp were analyzed by GUS reporter genes in T1 transgenic rice lines. The main results were as followings:1.The promoter of OsANKp was amplified by PCR using the specific primers of OsANKp and rice genomic DNA as template. The PCR products was sequenced and the cis-elements were analyzed by online software in PLACE website. The transgenic vector of OsANKp:: GUS was constructed and identified by sequencing, which was further transformed into rice by agrobacterium mediated method. The regenerated plants acquired were further subjected to PCR identification to get ride of the false transgenic plants and the seeds of the transgenic plants were harvested to get their T1 plants lines. The results of GUS expression analysis indicated that the promoter can be induced to express by mechanical wounding, Cnaphalocrocis medinalis and Magnaporthe grisea.2. The promoter of OsPITAp was amplified by PCR using the specific primers of OsPITAp and rice genomic DNA as template. The PCR products was sequenced and the cis-elements were analyzed by online software in PLACE website and more then 10 cis-elements in OsPITAp were found. The transgenic vector of OsPITAp:: GUS was constructed and identified by sequencing, which was further transformed into rice by agrobacterium mediated method. The regenerated plants acquired were further subjected to PCR identification to get ride of the false transgenic plants and the seeds of the transgenic plants were harvested to get their T1 plants lines. The results of promoter expression pattern analysis indicated that promoter failed to respond to mechanical wounding and Magnaporthe grisea infection, but responded to MeJA or NaCl treatment. On the other hand, an increased expression of GUS in buds was also observed when buds was disposed with INA or Bestatin.