| Taking vector of plasmid SCMVâ… -C2 gene and express promoter in maize, we transform sweet maize immature embryo and callus. At last obtain the transgenic plants successfully. The influence of Agrobacterium tumefaciens density, the length of co-cultivation period and temperature on transformation rate has been studied. The findings of study are as follows:1. When the temperature stays 25℃and the OD600 value of the bacterium solution are respectively 0.2, 0.4, 0.8and1.0, 1.4, the maize callus after 5 minutes infection undergoes co-cultivation, restorative cultivation and resistant callus screening at 22℃. It is found that the transformation rate is the highest and formulation rate of resistant callus reaches 23.7%. when the OD600 value of the original bacterium solution is 0.6 and the dilute density is 0.3.2. When the temperature stays 25℃and the OD600 value for original bacterium solution density and the dilute density being0.6 and 0.3, temperature of co-cultivation stays at 22℃, time of tincture are 2min, 5min, 10min, 15min each. It turns out that the transformation efficiency rate goes highest at time of 5min formulation rate of resistant callus.3. When the OD600 value for original bacterium solution density and the dilute density being0.6 and 0.3, temperature of co-cultivation stays at19℃, 220C, 25℃each, time of co-cultivation 1d, 2d, 3d each. It turns out that the transformation efficiency rate goes highest at time of co-cultivation 3d and temperature of co-cultivation at 22℃of formulation rate of resistant callus.4. Transgenic maize plants are obtained with maize immature embryo and callus through the mediation of Agrobacterium tumefaciens. An investigation of the transgenic maize molecules through PCR-detection and RT-PCR detection guarantees us the fact that SCMVâ… -C2 gene has been successfully incorporated into the maize genome. |
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