Foundation Of Regeneration System On Codonopsis Lanceolata Benth.et Hook. One Of Chinese Medical Herbs

Two regeneration systems were established respectively on Codonopsis lanceolata Benth.et Hook. through two approachs: one was inducement and differentiation of callus with leaf, radicles, stems and seeds as explants and the other was inducement of clustered shoot with axillary bud as explant. The main results can be summarized as follows:1. Inducement of callus: Callus could be induced by different explants including leaf, petiole, stem and seed, but induce ratio and growth form of callus were different from each other. It was proved that leaf was the best explant for inducing callus. The effects of different hormone combinations on callus formation were greatly, and auxin 2,4-D is a key factor in callus formation. The best culture medium was MS+2,4-D 0.5mg.L^-1+6-BA 1.0mg.L^-1+KT 0.5mg.L^-1, on which the highest induce ratio was 100%.2. Subculture of callus: Callus under different light conditions were both proliferated, but under 16h.d^-1 conditions, the callus multiplication times could reach to 9.5, and the callus grew fast and texture closely. This kind of subculture callus could be differentiated whit a high degree in the later differentiation process. The best subculture medium was MS+6-BA 0.75mg.L^-1+NAA 0.5mg.L^-1.3. Inducement and proliferation of buds: The callus turned green and formatted many buds after transferred into the culture medium, and then clustered shoot was formatted. Different light conditions affected significantly on the differentiation of the regenerated plants, the regenerated seedlings were robust whit more leaves under 16h.d^-1 condition while the regenerated seedlings were thin under 8h.d^-1 condition. The best medium for differentiation was 1/2MS adding 6-BA 0.5mg.L^-1 and NAA 0.2mg.L^-1 on which the highest differentiation rate could reach to 89.4%.4. Roots regeneration: Roots began to grow after the callus were transferred into the rooting medium.45 days later the growth of roots was stable. On the 1/2MS medium with IAA 0.2mg.L^-1 and NAA 0.2mg.L^-1, the highest rooting rate could reach to 92.7%, on which the number of roots was 26.8 and the length of roots could reach to 6.13cm.5. Inducement of clustered shoot and regeneration system was established: Shoot could induce with axilla bud as explant, and the best culture medium was 1/2MS adding 6-BA 2.0mg.L^-1 and NAA 1.5mg.L^-1, on which the highest induce ratio was 55.7%. The best culture medium for subculture was MS adding 6-BA 3.0mg.L^-1 and NAA 1.0mg.L^-1, and the highest differentiation time was up to 5.49 after subcultured two times. 1/2MS medium with no hormones was furthest suitable for root'induction.6. Refine and transplant the seedlings: The regeneration plants were hardened 7d in closed bottles natural light. Then opend the bottle caps and hardened 3d. The medium must be sterilized before transplanting. Early transplanting days, it is necessary to shady to full light intensity is about 30%. The best transplanting matrix was rotten quality soil, vermiculite and river sand by 1 to 1 to 1 and 82.5% seedlings could survive.