Research on tissue culture and plant regeneration for roots of Ficus hirta / 中草药

Objective: To solve the shortage problem of large-scale planting the seedlings in the roots of Ficus hirta by rapid propagation in vitro tissue culture. Methods: The MS and 1/2 MS media were used as basic media, The multi factor combination of plant growth regulator (6-BA, NAA, 2, 4-D, IAA, KT, and IBA) on the seedling subculture and root culture was studied by orthogonal design. Results: The best medium for the adventitious bud induction was MS + 6-BA 1.0 mg/L + NAA 0.3 mg/L, 72 adventitious buds were obtained by differentiation with 20 d induction of explants; The best medium for cluster inducing and subculture was MS + 1.0 mg/L 6-BA + 0.3 mg/L IAA + 0.3 mg/L KT; The best rooting medium was 1/2 MS + 1.0 mg/L IBA + 0.3 mg/L NAA, and the rooting rate was 100%. Being suitably Transplanted to peat-perlite (1:1) matrix, the 30 d survival rate was 93%. Conclusion: The tissue culture for the roots of F. hirta could be used to produce test-tube seedlings for large scale planting.

Tissue culture of medicinal plant Swertia mileensis / 中草药

Objective In order to preserve the natural resources of Swertia mileensis which has been destroyed seriously, the method of artificial propagation by way of tissue culture has been systematically studied. Methods The immature stems and the mature leaves were the best material explants in rapid propagation in all of the experiments. These explants were cultured on MS culture media by adding different portions of hormones at various cultural conditions. Results For the leaves, the suitable phytohormone combination to induce callus are Zt 0.5 mg/L+NAA 0.1 mg/L+IBA 0.1 mg/L or BA 0.5 mg/L+2,4-D 0.1 mg/L+IBA 0.1 mg/L; for the stems, they are BA 0.02 mg/L+Kt 0.04 mg/L+IBA 0.05 mg/L; for the adventitious buds, they are BA 2.0 mg/L+NAA 0.5 mg/L or BA 2.0 mg/L+IBA 0.1 mg/L; and for the roots, they are MS+Kt 0.01 mg/L+IBA 0.5 mg/L+NAA 1.0 mg/L. Conclusion Tissue culture of S. mileensis can make its propagation rapid, its resources preserved, and its utilization last.

Studies on industrialization cultivation of Dendrobium candidum / 中草药

Object To optimize the fitting culture media for multiplication of adventitious bud and ra-dication of Dendrobium candidum Wall. ex Lindl. and the length of adventitious bud for succesive transfer multiplication. Methods In the same culture condition, the adventitious bud with different length developed in the various culture media for multiplication and radication. Facing the different growth status, the diversity test and general analysis were carried on for them. Results There were significant difference in different length of adventitious bud, difference between the multiplication and radication of culture media. Conclusion The adventitious buds in (1.2?0.1) cm length are superior for succesive transfer multiplication, MS adding BA 1.0 mg/L, NAA 0.2 mg/L is the best media for the adventitious buds multiplication, 1/2 MS adding NAA 0.2 mg/L is the test media for radication.

Tissue culture of Swertia bimaculata / 中草药

Objective In order to protect the natural resources of Swertia bimaculata which has been destroyed seriously,the method of artificial propagation by way of tissue culture have been systematically studied.Methods The stems,leaves,and stems with buds which were from the seedlings germinated from the seeds on the initial medium were taken as explants.These explants were cultured on MS culture media by adding different portions of hormones at various cultural conditions.Results The stems were the best material in speeding propagation among the three explants.The proper initial medium for the stems was MS+ BA 0.5 mg/L+saccharose 3.0%,the optimum medium for proliferation was MS+BA 0.5 mg/L+IBA 0.1 mg/L+saccharose 3.0%,and the best medium for rooting was 1/2MS+NAA 0.5 mg/L+saccharose 1.5%.Conclusion Tissue culture of S.bimaculata could make its propagation rapid,its resources preserved,and its utilization last.

Adventitious bud induction and endogenous hormones changing of lamina from Zanthoxylum dissitum in vitro / 中草药

Objective To establish a regenerative system in vitro and determine the endogenous hormones of Zanthoxylum dissitum. Methods Lamina of Z. dissitum was used as the explant. Callus adventitious bud differentiation was carried out by culturing on MS with different hormones. At the same time,in the course of callus induction,four endogenous hormone of GA3,IAA,ABA,and ZR were determined by ELISA. Results The optimum medium for adventitious bud regeneration of Z. dissitum was MS+6-BA 0.5 mg/L+NAA 0.1 mg/L,and the highest induction rate could reach to 61%. During the process of the differentiation of explants callus,the contents of ZR and IAA were correspondingly higher than other endogenous hormones,the content of ABA was always kept in low level during this process,the content of GA3 was kept in the trend of upgrading during the earlier differentiation period. Conclusion It could be considered that ZR and IAA should be the critical factors in the bud induction. According to the results,a proper adding of GA3in the culture medium could improve the differentiation rate of the adventitious bud. On the other side,ABA might be the negative regulation factor.

Condition for induction and differentiation of callus and propagation of adventitious buds in Bupleurum chinese / 中草药

Objective To study the condition for induction and differentiation of callus and propagation of adventitious buds in lamina,stem,and bud of Bupleurum chinese and establish a new method for rapid propagation.Methods In MS media added with different phytohormones,calli were induced from explants of lamina,stem,and floral bud of B.chinese,adventitious buds and adventitious roots were differentiated from calli of stem and floral bud,test-tube plantlets were formed.Results MS Medium added with 2,4-D 1.0 mg/L,KT 0.5 mg/L,and 6-BA 0.5 mg/L was suitable for calli induction of the lamina,stems,and floral buds.In medium added with 6-BA 1.0 mg/L,NAA 0.03 mg/L,CM 15% and CH 500 mg/L,the differentiation rate of floral buds callus was the highest.MS Medium added with 6-BA 1.5 mg/L,NAA 0.05 mg/L and CH 250 mg/L was suitable for propagation of test-tube plantlets,1/2 MS medium added with NAA 0.5 mg/L was suitable for rooting.Conclusion A great deal of test-tube plantlets could be differentiated and propagated rapidly by calli induced from stems and floral buds of B.chinese.Then the regeneration plantlets with normal growth and development are obtained.

Tissue culture system of Cynanchum otophylum in genetic transformation / 中草药

Objective To establish the tissue culture system of Cynanchum otophylum in genetic transformation. Methods Asepsis seedling was set up by using different explant parts, disinfectors, and disinfecting time. The callus was induced, breeded, and differentiated by using different media, different hormone categories and combinations, and different hormone concentrations of 2,4-D and KT. Results It was the best method that asepsis seeding was built up by using seed as the explant and using 10% NaClO to treat the seed for 20—30 min or using 0.2% HgCl2 to treat the stem for 3 min. The seed, root, stem, and leaf of C. otophylum can form callus easily. The 2,4-D was important in forming callus. C. otophylum was more sensitivity to KT. The effect was better when KT was used to induce callus or adventitious buds. MS+2,4-D 1.0 mg/L+KT 1.0 mg/L was the better medium to induce callus to form. The callus cant be induced to form adventitous buds without reference to using seed or root, stem, and leaf as the explant, without reference to using hormone 6-BA or KT, ZT, 2ip. Gentamicin 100 ?g/mL is the optimum pressure in genetic transformation of C. otophylum. Conclusion The seed or stem is the ideal material bombarded by particle gun that is induced to form callus for 14 d in the media MS+2, 4-D 1.0 mg/L+KT 1.0 mg/L and without forming adventitious buds.