Preliminary Study Of NPR1Transcriptional Level Under The Control Of Synthetic Promoter On Broad Spectrum Resistance Of Arabidopsis Thaliana

High-level expression of exogenous resistance gene that regulate by synthetic pathogen-inducible promoter is one of the important means to improve crop resistance to disease. Therefore, improve the disease resistance by transgenic technology should overcome the two questions that how to select and precise control the exogenous gene expression.A WGFS-mini35S synthetic pathogen-inducible promoter was obtained using the dipolymers such as2×F-box,2×S-box.2x GST1-box and2×W-box and35S minimal promoter. Based on this, the results are as follows:1. Evaluated the stability and applicability of four reference genes (RTEF、RARS、 EACT、AACT) using E. urophylla×E. grandis leaf. Eucalipu pellida leaf; Eucalyptus urophylla leaf and green callus, red callus, white callus of Eucalyptus urophylla as materials.2. Transient expression analyzed the induced activity of synthetic pathogen-inducible promoter using Eucalyptus urophylla leaf as materials. The result showed that the background activity of synthetic pathogen-inducible promoter is0.2951times to35S promoter. The induced activity of synthetic pathogen-inducible promoter increased to2.36times of35S promoter,7.87times of background activity when treated with Phytophthora spore. The induced activity of synthetic pathogen-inducible promoter increased when他reated with SA.3. Successfully constructed the WGFS-mini35S-nprl expression vector and transformed it into Agrobacterium tumefaciens.4. Two types of transgenic Arabidopsis thaliana was obtained using Agrobacterium tumefaciens-floral dip method transformed WGFS-mini35S-nprl expression vector and35S-nprl expression vector(preserved in our laboratory)into wild type Arabidopsis thaliana.5. Morphological observation showed that WGFS-mini35S transgenic Arabidopsis thaliana root length is longer than35S-nprl transgenic Arabidopsis thaliana. and lateral root developed. 6. QPCR analysis of wild Arabidopsis thaliana,35S and WGFS-mini35S transgenic Arabidopsis thaliana treated with SA and WGFS-mini35S transgenic Arabidopsis thaliana treated with PC and RS; Relative expression of35S and WGFS-mini35S were2.99times and6times to CK,respectively; Npr1relative expression of wild Arabidopsis thaliana,35S transgenic Arabidopsis thaliana and WGFS-mini35S transgenic Arabidopsis thaliana were2.56times.5.67times and14.31times to CK treated with SA.1.96and2.43. NPR1relative expression of WGFS-mini35S transgenic Arabidopsis thaliana were1.96times and2.43times to CK treated with PC and RS. respectively.All the results above showed the induced activity of WGFS-mini35S promoter was higher than35S promoter.