Small Cluster Rhodiola Tissue Culture Research

Rhodiola dumulosa(Franch.)S.H.Fu is a plant that belongs to Rhodiola L. of Crassulaceae. The root and stem of the plant can be used as medicine to anti-hypoxia, antigatigue, chill-proof and immunoregulation. It has been widely used in traditional Chinese medicine.In this paper, Rhodiola dumulosa stems were used as explants, and discussed the inductive effect of callus, adventitious bud and root in different plant growth regulators and different concentration combination culture. The system of tissue culture of Rhodiola dumulosa was established. The changes of polysaccharide and POD enzyme in different callus were also researched.1. The best callus induction medium was MS with 0.85 mg/L 6-BA 0.34mg/L NAA and 0.33 mg/L 2,4-D, in which the rate of the callus induction can reach 90%.2. The state of callus is different on different medium. The callus is tight and yellow on the medium with TDZ and 2,4-D; The callus is tight and green on the medium with TDZ and 2,4-D; The callus is yellow and white on the medium with 6-BA and 2,4-D; The best callus proliferation medium was MS with 0.5 mg/L TDZ and 2.0 mg/LNAA, in which the callus had 47.26 times proliferation rate.3. Adventitious bud induction medium was MS with 1.0 mg/L 6-BA and 0.5 mg/L NAA or with 1.0mg/L TDZ and 0.5mg/L NAA. However, the induction rate was very low. The adventitious buds can be with higher petioles and bigger leaves on the medium which is MS with TDZ.4. The best medium of rooting induction for adventitious buds was MS+IBA 1.0mg/L medium and the rooting rate was 100%. The average root number was 10-15 and average root length was 2-3cm. The regeneration plant has been transplanted after rooting and the survival rate was 100%.5. The polyacrylamide gel electrophoresis was used to analysis POD enzyme of callus and plant on different culture medium. The result shows that POD involved in the morphogenesis in some degree. The content of POD rise up when the callus is going to die. The callus which can induct adventitious bud have special POD striping.6. Concentration of the polysaccharide from the callus on five different medium with the best proliferation is being analyzed. The results shows that callus on the medium of MS+TDZ0.5mol/L+NAA2.0mol/L contains the highest concentration of polysaccharide, which reached 2.15%,1.93 times to the wild plant.The polysaccharide extractive form callus on MS+TDZ0.5mg/L +NAA2.0mg/L, MS+6-BA0.5mg/L+2,4-D0.5mg/L have the better antioxidant activity.