Optimization Of Transformation Tissue Culture System Of Xinjiang Muskmelon And Transforming Anti-fungal Genes

Using the 3 days of Xinjiang muskmelon cotyledon as explants, regenerated plantlets were induced on basic MS medium supplement with different hormones. The experiment was divided into 8 parts.(l)The optimum 6-BA density was 1.0mg/L in tested 0.5, l.0, 1.5, 2.0 mg/L 6-BA. The highest frequency shoot regeneration observed was 93.3%. Many shoots were induced near the hypocotyls, showing certain polarity. (2) On basis of the 1.0 mg/L 6-BA, the optimum IBA density was 0.1 mg/L. The explants obviously expanded.(3) Using 0.3-0.5 mg/L GA can not only accelerate the growth of the regenerated shoots but also help to the surrounding germinal growth.(4) Using 0.2 mg/L mannitol can enhance the rate of transformation. (5) To restrain the Agrobacterium tumefaciens, Cerb and Amp was used. The effect of 500 mg /L Amp was better than Cerb's. (6) the optimum time of transform was 8 mins dip-dye and 3-5 days cultured explants with agrobacterium.On the basis of regeneration protocol of Xinjiang muskmelon,two genes chitinase and β -1.3-glucanase were transferred into melonby Agrobacterium tumefaciens. Then the explants were selected by75mg/L kanamycin in the medium. After obtaining the resistanceexplants, using the way of PCR proved the aimed genes whether hadsuccessfully infected. The expression of the two enzymes protein wasobserved by SDS-PAGE. (7) the PCR result showed chitinase gene andβ -1.3-glucanase gene had been trans- formed in melon. (8) Observingthe expression of the chitinase by SDS-PAGE, but not the β-1.3-glucanase.