The Interaction Protein With Fusarium Graminearum's Effector Xylanase In Wheat And Its Characterization Analysis

Fusarium graminearum is the major Phytopathogenic fungus for wheat head blight.Fusarium graminearum secretes a battery of cell wall degrading enzymes(CWDE)to help its infection,and xylanase is one of those.Further more,xylanases are important effectors of Fusarium graminearum,and they play key roles in interfering with the wheat's resistance to the fungus.The wheat mounts innate immune response when it sense pathogen's PAMPs to block its infection or spread.The wheat also secretes series of cell wall degeneration enzyme inhibitor proteins to combat the CWDE and activates the immune reaction.So it's of great importance to find out the host target proteins in the wheat that intereacts with xylanase.to identify the functional domain involved in protein-protein interaction and to analyze the expression level of target proteins under pathogen infection.(1)Using FG11304 as bait,a chitinase fragment was screened in wheat cDNA expressing library by yeast two-hybrid.(2)The full length of the chitinase gene was isolated based on flanking sequences of the gene sequence on NCBI.The coding region of the gene was fused with AD domain in pGADT7 vector.The interaction of the chitinase with FG11304 was confirmed by co-transformation of FG11304 and chitinase-pGADT7 vectors in AH109 yeast strain.Their interaction in plant cells was further confirmed by BiFC assay.(3)Used bioinformatics method to analysis the chitinase gene's function domain and constructed series short vectors to find out chitinase's fuctional domain intereacting with xylanase FG11304.(4)Semi-quantitative RT-PCR found out that the expression of chitinase was induced in wheat spikes by Fusarium graminearum,and the expression in leaves is higher than in spikes...