Screening For Arabidopsis Thaliana Mutants With Enhanced Resistance To Sclerotinia Sclerotiorum And Their Molecular Characterization

Sclerotinia sclerotiorum is a ubiquitous phytopathogenic Ascomycete fungus, which is capable of infecting a wide range of plants. It causes serious diseases in some economic plants and has become a limiting factor for production. However the mechanism underlying the plant-fungus interaction remains mostly unknown. We here used Arabidopsis thaliana as a good material to screen for T-DNA activation tagging mutants with enhanced resistance to the Sclerotinia disease. Then we cloned the genes which may be associated to resistance, constructed the overexpression vector and transformed Arabidopsis to test functions of these candidate genes.The major results were as followed.1 Obtained 2 mutants with enhanced resistance to S. sclerotiorum. by screening 50,000activation-tagged mutants .2. Cloned the genes associated to resistance to S. sclerotiorum which obtained by SSH andconstructed the overexperssion vectors, and then transformed A. thaliana' s wide type.3 Cloned the genes associated to oxalic acid which obtained by Tail-PCR and constructed theoverexperssion vectors, and then transformed A. thaliana's wide type.4. Obtained 2 pure mutants according to screening with Basta resistance with T₃ transformed gene mutants.5. The RT-PCR results confirmed these candidate genes overexpressing in plants.In summary, our work will facilitate to searching the mutants resistant to Sclerotinia disease and dissecting for molecular mechanism of Arabidopsis -Sclerotinia interaction.