Résumé
Objective To establish an efficient tissue culture and rapid propagation system, and study the protocorm proliferation and regeneration conditions using seeds as explants in Dendrobium officinale. Methods Seeds of D. officinale were used as explants, protocorm was induced on inducement medium. After proliferation, the protocorm were transferred to the regeneration medium. Then the regenerated shoots were transferred into rooting medium to induce rooting of plantlets, and developing complete plant. Results The basic culture medium for D. officinale growth was 1/2 MS; The best culture medium formula for inducing protocorms was 1/2 MS +1.0 mg/L 6-BA + 0.2 mg/L NAA + 50 g/L mashed potato; The optimal proliferation medium for protocorm was 1/2 MS + 1.0 mg/L 6-BA + 0.5 mg/L NAA, and the maximum multiplication factor could reach 23. The optimal regeneration medium was 1/2 MS + 2.0 mg/L 6-BA + 0.2 mg/L NAA, regeneration rate can reach 95%; The best culture medium for seedling rooting was 1/2 MS + 0.3 mg/L NAA + 50 g/L potato juice, and rooting rate reached 100%. Conclusion This research provides an effective way for keeping good varieties of features and rapid propagation of D. officinale, at the same time helps to solve some theoretical problems in factory production of D. officinale.
Résumé
Objective: To establish the introduction and culture system of the hairy roots in Solanum lyratum and to screen the clone of hairy roots with more diosgenin. Methods: The explants of S. lyratum were infected by Agrobacterium tumefaciens strain C58C1, to obtain the hairy roots and construct the genetic transformation system of the hairy roots in S. lyratum. HPLC was used to determine the diosgenin in the hairy roots. Results: The optimum transformation results were obtained with the max inductivity of hairy roots of 83.33% during infecting time for 10 min by C58C1 and co-cultural time of 4 d. The average content of diosgenin in the hairy roots was 4.620 mg/g, it was 2.652 times as high as that in the leaves (1.742 mg/g) which had the highest diosgenin content in the different tissues of wild type plant of S. lyratum. Conclusion: It is an effective way to obtain diosgenin from the hairy roots of S. lyratum.