Study On Transformation Of Sunflower (Helianthus Annuus L.) With Oxalate Oxidase Gene By Agrobactrium Tumefaciens

Helianthus annuus L. is a very important commercial crop in Inner Mongolia, its seeds are rich in oil and have important commercial merits. The production of Helianthus annuus L. is jeopardized seriously by S. sclerotiorum day by day, Its antigen has not been found so far. It is a fundamental way to seek control method based on the determinant factor of pathogenesis. Aiming at oxalic acid, the major pathogenicity factor of S. sclerotiorum, the introduction and expression of genes encoding enzymes that degrade oxalic acid is a potential powerful strategy to enhance resistance to the fungi that secrete oxalic acid by using Agrobacterium tumefaciens harbouring the plasmid containing OxO from wheat. The main results were as follows:The regeneration system has been established successfully. The main results are as follows:1 Differentiation medium: MS + 0.5mg/L 6-BA + 0.25 mg/L IAA + 0.1mg/LGA, pH 5.82 Coculture Medium: differentiation medium+100μmol/LAS,pH 5.83 Selective medium: differentiation medium + 50mg/L Kan + 500mg/L Cef,pH 5.84 Root-inducing medium: 1/2 MS + 25mg/L Kan + 300mg/L Cef + 0.5mg/L NAA,pH 5.8OD6oofor bacterium concentration inoculation is about 0.3～0.4, the suitable inoculation time is within 9 minutes～10 minutes,then cocultured for 3 days. Shoot apice was used for OxO agrobacterium-mediated transformation. 14 regenerated Kanamycin resistant plants were obtained from the transformation above, Two plants showed specific positive band which is the same as the plasmid control by PCR reaction.Transformation frequency is 0.4%.