Studies On Establishment Of Tissue Culture And Cell Suspension Culture Systems Of Codonopsis Vinciflora Kom.

Codonopsis vinciflora Kom. is a perennial herbaceous plant in the Campanulaceae family and mainly distributes in regions of mountainous, grassland or bush at the elevation of 3000～4600 m in the central and south-eastern Tibet, Yunnan northwest, western Sichuan and other places. As a traditional medicine plant, Codonopsis vinciflora is unique for China and has very high medicinal value. It contains saponins, trace alkaloid, sucrose, glucose, inulin, starch, mucus resin. Fortifying its roots, mainly for the expelling the heat, cold governance, rule of tuberculosis, gastritis, and other diseases, cough and expectoration, the pain, stomach tonic effect. This article mainly discussed how to establish a plant regeneration system of Codonopsis vinciflor Kom. and cell suspension. The results provided fundamental supports to induction, domestication and culture of seedlings at a factorial scale, and to studies on medicinal ingredients of Codonopsis vinciflora Kom. The main results were as follows:When treated with 0.1% mercuric chloride for 3 minutes, the earthnut could be in the best disinfection situation; for the sterilization of the stem, 0.1% mercuric chloride treating for 30 seconds could bring the better pollution rate and survival rate relatively . If treated in the 84 disinfectant, stem in the 84 concentration of 6% for 10 minutes showed the best effect. As for the leaf disinfection, the disinfection time was 15 seconds, the pollution rate and the survival rate were better.The 6-BA and NAA combination were used to induce the adventitous buds-regeneration of Codonopsis vinciflora Kom. The result showed : the tender stem were more easily induced to sprout adventitious bud . With the explants of stems, the induction rate of adventitious buds on the medium containing 6-BA3mg/L and NAA1mg/L reached 86.7%. Averagely one explant could sprout 19.8 adventitious buds; regenerating culture medium is MS+6-BA0.5mg/L+NAA1.0mg/L. 1/2MS added the NAA,IBA, 2,4-D respectively and various concentrations of the three phytohormone, and the medium added activated carbon failed to induce root. It indecatecd that it was difficult for the rooting of adventitious buds.It showed that leaf explants were greater than earthnuts and stems as for the capability of inducing callus. Adding 3 mg/L 6-BA and 0.2mg/L2,4-D to the MS culture medium ,the inducing rate could run up to 100%.On culture medium MS + NAA 0.5 mg/L+6-BA2.0mg L, the rate of callus inducing into buds is the highest, reaching 87.4%. When cultured the callus suspension cell ,the medium was still MS +2,4-D 0.2 mg /L+6-BA3.0mg /L, and the growth of callus was the most remarkable, with 2g callus growing up to be 8.58g fresh weight after 20 days of inoculation.