| By bombardment transformation we have successfully intergrated the AHAS gene of PCD220 plasmid into maize calli genome and regenerated transgenic plants of resisant-chulorsulfuron and studied some factors affecting transformation efficiency in particle-bombardment system of maize.Being cultured on the same 2,4-D concentration ,calli induction ratios are different among various genotype, and embryo age so on, When embryo calli of maize was used to be the receiever in particle-bombardment transformation. It is easy to induce embryo calli about ten days embryo; Otherwise it is not easy to do.2,4-D concentration of medium has great efffect on maize calli induction . The great induction ratios of calli on different genotype need various concentrations of 2,4-D, generally it is from 0.5 to l.Oppm.During particle-bombardment transformation ,the times of bombardment and the concentration of golden powder and the shoot-distance will affect the transformation efficiency of calli. Considering the transformation efficiency ,we think it is suitiable that the golden powder concentration is lOOug/P and the times of bombardment is twice and the shoot-distance is 9cm.The properly concentration of 6-BA is different according to various genotype when calli split up,the suitiable concentrations of Q319 and 515 are lppm,and Iuyuan341 is 0.5ppm.PCR analysis on transformation plants demonstrated that it is positive ,and Southern blotting of trangenic plants has enormously hybrid hint,But not the negative control of untransformed plants .These results indicated that the AHAS gene on resistant-chlorsulfuron had integrated into maize genome. We have get the seeds of T0 by inhbreed on transgenic plants. |
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