| In this experiment the seeds of A. teptocarpa were used as initial culture materials to study the in vitro rapid propagation and industrial production model of A. teptocarpa. This experiment compared the effects of different pre-treating ways and sterilizing ways on aseptic germination of the seeds, and screened the optimized media and culture conditions in different stages of primary culture, subculture and rooting culture. Moreover, this experiment also explored the countermeasures and culture media of improving survival rates of transplanting, and successfully established in vitro rapid propagation technical system and industrial production model of A. teptocarpa. and it also supplied scientific gist for the large scale of industrialized production on A. teptocarpa. As follows were the main results of the research:(1) The pre-treating ways of seeds and the primary culture of A. teptocarpa. The pre-treating ways of the seeds were as follows:using 98% H2SO4 to soak the seeds for 25 min, and then socking the seeds in boiled water for 20 min. After that, the seeds were ready for being sterilized with 0.1% HgCl2 for 20 min, and then transferred to asepsis water for being washed 4-5 times. The sterilized seeds were inoculated on 1/2MS medium. The germinating percentage of the seeds was up to 86.4% after 15 days.(2) The proliferation culture of A. teptocarpa. MS medium added with 0.5mg/L BA and 30g/L sugar as well as 6g/L agar (pH 5.4-5.8) was available for the induction and multiplication culture. The culture temperature was about 25±2℃, light intensity was 1500 Lx, and illuminating time was 12h/d. After being cultured for 45 days, the multiplication coefficient was up to 2.74, and the seedlings were strong with dark green leaves, which were suitable for subculture and rooting culture.(3) The rooting culture of A. teptocarpa. The preferable culture medium for root inducing was 1/2MS medium added with 1.0 mg/L IBA and 30g/L sugar as well as 6g/L agar (pH 5.4-5.8). The temperature was about 25±2℃, light intensity was 1500 Lx, and illuminating time was 12h/d. Under the conditions the rooting rate could be up to 100%; and the quantity and quality of the roots was preferable. The roots could form favorable root systems after being cultured for 40 days so as to be transplanted into the nursery grounds.(4) The plantlets transplanting of A. teptocarpa. The preferable transplanting medium for the plantlets was the prepared mixed medium of fine sands, garden soils and vermiculites (1:1:1), which brought preferable transplanting effects with the survivable rate of 80%.(5) The establishment of the industrialized in vitro propagation model of A. teptocarpa. The industrialized in vitro propagation model was successfully established, and the main technical parameters were as follows:the proliferation coefficient was up to 2.74, the subculture cycle was 45 days, and the subculture time was 8 times every year; the rooting rate was up to 100%, and the survival rate of transplanting was 83.3%. In theory,100 aseptic plantlets germinating from the seeds of A. teptocarpa each year could propagate about 318000 plantlets, which could achieve the demand of in vitro rapid propagation. |
