Agrobacterium-mediated Transformation Of Gala Apple With APX Gene

Ascorbate acts as an important antioxidants in both enzymatic and non-enzymatic reactions in plant cells. Under the adverse circumstance forces, plants can produce massive ROS which has very strongly poisonous function to the plant. Ascorbate peroxidase is the critical enzyme of scavenging the reactive oxygen species. When the reactive oxygen species were accumulated, the APX expression and detoxification of H2O2 through reaction of ascorbate and H2O2 can be immediately induced. Researchers hope to enhance the ROS elimination enzyme activity through the genetic engineering method. Thus sharpens the ability of the anti-oxidation forces and it would be also helpful to get more information about the role of APX playing in AsA-GSH cycle and its metabolism pathway. In this research, gala apple with APX gene was acquired by Agrobacterium tumefacien to lay a foundation for breeding the variety which has the high resistance.The main results were as follows:1. Agrobacterium-mediated genetic transformation system was optimized, and the transgenic plantlets of apple cultivar carrying APX gene were obtained.Factors afecting the transformation frequency were studied, and some parameters were determinated. The results indicated that better transformation rate was gained when the leaves was precultivated 2 days, the OD600 of grobacterium was about 0.6, Agrobacterium infected 10 min, and 10 days of detention screening.2. The better selective efficiency was gotten when the concentration of hygromycin was 3 mg/L Hyg.3. Result s of PCR and Gus proved that the APX gene had integrated into apple genome.