Study On In Vitro Rapid Propagation Technique Of Prunus Triloba Var. Atropurpurea

In this experiment, we taken Prunus triloba var. atropurpurea as the test material, and studied several key issues in the process of in vitro rapid propagation and tissue culture. We selected 3 different parts of the Prunus triloba var. atropurpurea annual branch terminal buds, axillary bud, and stem segments as explants. Through the induction of callus, adventitious buds and adventitious buds in vitro, this article discussed the effects of different basic medium, different plant hormone types and concentration on the rapid propagation of Prunus triloba var. atropurpurea in vitro. The results were as follows:1. During the sterilization of explants, the optimum method of sterilization for terminal bud was 75% C2H5OH(5s) + 0.1% HgCl2(4 min); the optimum method of sterilization for axillary bud was 75% C2H5OH(10s) + 0.1% HgCl2(4 min); the optimum method of sterilization for annual stem segment was 75% C2H5OH(15s) + 0.1% HgCl2(6 min). When the stem segments of hydroponics though sterilization with 75% C2H5OH(15s) + 0.1% HgCl2(5min), the experiment found that it could reduce the rate of explant contamination.2. The three explants of Prunus triloba var. atropurpurea with MS as the basic culture medium had the best effect of start-up. Explant buds, axillary bud, stem segment start rates were 95%, 96%, 93%, and the growth condition was good. The best combination of starting medium was MS+6-BA 2.0 mg/L+NAA 0.2 mg/L. Compared with the three explants, the germination rate of stems was the highest. And June was the optimum time for the explant sampling of Prunus triloba var. atropurpurea.3. In the following generation of Prunus triloba var. atropurpurea, it was mainly proliferated in two ways. One method was to induce adventitious buds proliferation through callus, and it could induced more adventitious buds by using 1/2MS+6-BA 2.0 mg/L+NAA 0.1 mg/L as medium. The other method was to induce the stem buds for clusters of adventitious buds proliferation, and the optimal proliferation medium was MS+6-BA 2.0 mg/L+KT 1.0 mg/L +NAA 0.1 mg/L. The 25 day culture period could achieve the maximum proliferation efficiency, and the proliferation rate was 92.19% and the proliferation coefficient was 4.27, and cluster bud growth state health. The number of switching should not be more than 6 times.4. By using different types and concentration of auxin to root culture for the tissue culture seedling of Prunus triloba var. atropurpurea, the effect was significant difference. The optimum rooting medium was 1/2 MS+IBA 0.2 mg/L, and rooting rate was 87.47%, and adventitious root growth was better. The survival rate of tissue culture seedling that was cultivated in the stroma of V(garden soil): V(peat): V(perlite) = 1:1:1 was the highest, 78.62%. Seedlings grew healthy and fast, and the leaves were green. When the root length of the adventitious root was selected to be 3-5cm, the survival rate of seedlings was the highest and it was 81.33%.5. The browning rate of Prunus triloba var. atropurpurea callus could be reduced by adding different anti browning agents to the medium. When the concentration of AC was 0.2-0.3 g/L, the inhibition ability of Callus Browning was the best.6. By adding one or more antibiotic combination to the culture medium, it has a significant effect on the tissue culture seedling. The penicillin sodium added only to the tissue culture seedlings had the best antibacterial effect. The penicillin sodium 300 mg/L + streptomycin 100 mg/L that used the antibiotic combination had the best antibacterial effect.