The Study Of Rhododendron Parvifolium Adoms On Tissue Culture

Rhododendron Parvifolium Adoms, evergreen arboret, is a kind of rhododendron plants in ericaceae. As an excellent undomesticated flower specie in Changbai mountain, it is the third key protected plants in our country with its rare amount. Aiming to protect and develop the resource of Rhododendron Parvifolium Adoms, this experiment has studied the explants on its sterilization and implantation, start training, subculture, callus induced and increase, differential and seedling of callus, root forming training and acclimatization. Trying to select the best culture medium and conditions, confirmate the suited method of tissue culture for the Rhododendron Parvifolium Adoms. And hopes to give technology support for transgene and cell fusion. The main results are as follows:1)The best tissue culture environment is:temperature25±2℃,light 24h/D,light intensity 20001x,pH5.0-5.5.When the pH is 5.0,the spire grows faster which is benefitted for proliferation culture. When the pH is 5.5, the spire becomes stronger which is benefitted for vigorous culture.2)The time for explants collected:the current year young shoots collected at July is the best; stem is better than stem tip because of its more quickly start-up and higher rate and lower pollution.3)The sterilization method of explants:use 0.1%HgCl2 for sterilizing and maintain 2.5min; it is better for the sterilization effect with tween 1-2 drops in.4)The start-up culture:use Read+ZT 0.5mg/L+NAA 0.05mg/L as medium, pH is 5.0,sucrose is 30g/L,agar is 12g/L,and the induction rate and growth is best; cultivate for 40 days, the start-up rate of explants is 62.96%.5)The proliferation culture:use Read+ZT 0.5mg/L+NAA 0.05mg/L as medium, the per proliferate multiple is 5, buds can be diversified from basal stem, the proliferate shoots grow stronger. Enhancing more light intensity, setting the temperature difference between day and night, and increasing the nutrients concentration in the medium, are helpful for cultivating stronger proliferate shoots.6)The seedling culture:use Read+NAA 0.2mg/L+AC2g/L as medium, the effect is better. The leaves is bigger and green and growing stronger. What's more, this medium can promote root forming.7)Root forming culture:Read+NAA 0.2mg/L+AC2g/L medium can reach the best effect, and the rate is 95.23%;tissue shoots are liked to form roots, their growth are coincide and the survival rate is higher. With this medium, Rhododendron Parvifolium Adoms grows quickly and stronger, so that it can be cut to proliferate and the proliferate multiple is 4.Using stem for proliferation variation is rarely appeared, meanwhile, the tissue culture process can be simplified because of the effects of seedling culture, root forming and proliferation caused by this medium.8)Callus culture:the effect of callus induced is better on the medium of Read + 2,4-D 1.5mg/L+NAA 0.1mg/L;the rate of callus induced about leaves is higher than stem tips; the proliferation of callus is fitted on the medium of Read +ZT 3.0mg/L+NAA 0.1mg/L;the Read+ ZT0.8mg/L+NAA 0.08mg/L medium is fit for the callus differentiation culture.9)Acclimatization: the process of acclimatization should be carried out at spring or autumn with suitable temperature and humidity, and the survival rate in spring is higher than autumn; on the matrix of the ratio with garden soil:turf: humus=1:1:1 the survival rate is highest; transplant work should be done under half light environment.10)Rooting of cutting: half light environment is better for this work; the survival rate of rooting growth is better on the matrix of moss; the rooting treatment is necessary.