Abstract:

Abstract: Selecting opening wild poppy bud as receptors , plasmid pBIAFP containing the radish peptide AFP (antifungal protein) gene as donor, in different concentrations of cane sugar solution, adding fresh pollen and plasmid DNA containing the target gene , shocking with SKH250LH-type ultrasonic oscillator for treatment pollen processing fluid. After all these process, we observed its pollen germination under OLYPUS light microscope with Coolsnap at different times. After different pollination time, dropping the pollen on wild poppy stigma supplemented with artificial method, bagging for seed and measuring seed-setting rate. The results show that, after transformation of radish peptide with pollen-mediated method , 11% sucrose concentration and 10-15h treatment is best for wild poppy pollen germination , when the sucrose solution concentration is 8%, 5h can get better seed-setting rate. After future kanamycin selection and PCR detection , we verified that exogenous AFP genes integrate into the genome of wild poppy initially and obtained transgenic T1 generation .It provide a new approach for special Chinese herbal medicine breeding .