| The calli were induced from immature embryos of four maize lines (Q31×Z31,Q31×Z3,Z3 and B73), analyzed effects of different factors on the rate of calli induction and regeneration. CHI gene was transferred into Embryogenic calli by the Agrobactrium-madiated and gene gun, studyed genotypes and other factors on rates of transformation.The main results were showed as following:1. Different genotypes showed different ability of emberyogenic calli inducing.B73 had most high rate of induction, Q31×Z31 and Q31×Z3 were high,but Z3 had little rate of it. The concentration of 2,4-D were most important factor on induction and culture of calli.2.0mg/L2,4-D was found to be most favorable for the induction of embryogenic calli,and our experiment had proved this point too. The addition of silver nitrate (25μM) also supported an increased frequency of embry-ogenesis.In process of differentiation of embryogenic calli,1.0mg/L6-BA could promote the differentiation of embryogenic calli.2. After the emberyogenic calli was transforred by Agrobacterium tumefaciens,we analyzed the effect of genotypes and other factor on transformation,the results showing that ,B73 had the most high transformation effiencicy of calli ,and the best transformation conditions were that bacterial concentration/OD600 was 0.4-0.6, infecting time was 10 min,co-culture temperature was 23℃.3. Studyed effects of genotypes and the combination of helium pressure and bombardment distance on transformation effiencicy of calli by gene gun, the result was that B73 had the most high transformation effiencicy of calli, 1100psi and 6cm were the best combination.4.The results of PCR and Gus staining analysis of transformants indicated that exogenes have been inserted into maize genome.The experiment also proved that some transformants have received Basta-resistance. |
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