Study On Transformation Of Apple With Antifungal Gene

Fungal diseases especially apple Alternaria Leaf Spot,Marssonina coronaria(Ell.et Daris)Daris,Physalosporapiricola,and Valsa mali are the major diseases on apple, and has become a major limiting factor on apple production. The genetic engineering of apple for disease resistance becomes an important issue. Synergistic action of plant defense genes appears to be an effective avenue in antifungal genetic engineering of plant. In this article, factors affecting buds regeneration from apple leaves were studied.Regeneration system of Gala leaves in vitro and NagafuNo.2 internode segments was optimized. By studying factors affecting agrobacterium-mediatedgenetic transformation, the agrobacterium-mediated genetic transformation syste-m of Gala was established. And theβ-1,3-glucanase and chitinase genes were introduced into Gala by means of leaf discs and finally got the objective seed-lings of Gala. The main results were as follows:1. High efficient and steady regeneration system of apple leaves in vitro was obtained. By adjusting the factors such as culture medium, concentration of growth hormone, dark incubation period, leaves from different parts and leaf polarity, established high efficient regeneration system of Gala and NagafuNo.2. On MS medium containing BA 5.0 mg·L-1+ NAA 0.2 mg·L-1 or TDZ 4.0 mg·L-1+NAA 0.6 mg·L-1 with abaxial surface of Gala leaves, adaixial surface of NagafuNo.2 leaves touching medium and after about 21d dark incubation treatment, the regeneration efficiency of Gala and NagafuNo.2 leaves reached 100% and 60%, respectively.2. The regeneration buds from NagafuNo.2 etiolation segments were obtained. By studying the effect of factors such as culture medium, concentration of hormone, the regeneration buds from NagafuNo.2 etiolation segments were got. Regeneration frequency was 83%, which was higher than leaves. A high regeneration frequency system was established.3. By studying effects of different antibiotics on regeneration of apple organs in vitro and the growth of adventitious shoots, the optimal selecting pressure was ensured.