Studies On Regeneration System Of Hypericum Perforatum L. And Expression Of Hyp Gene

Hypericum perforatum L., is a medicinally important plant used in the treatment of depression and virus. Hypericin is the major active constitutent. This paper using Hypericum perforatum L. as material, established its tissue culture and plant regeneration system, which will contribute to the large-scale extraction of hypericin; Expression of Hyp gene was also studied, hoping to find a new way to obtain large amounts of hypericin.The main results were as follows:The effects of basal medium, different kinds and concentrations of auxins and cytokinins on tissue culture and rapid propagation of Hypericum perforatum L. were studied. The results were as follows: MS medium supplemented with IBA 0.1 mg/L and 6-BA 0.1 mg/L was most suitable for the rapid propagation of Hypericum perforatum L.. The rooting rate reached above 90 % with 0.05~0.08 mg/L IAA on MS medium. The hypericin content from the tube seedling and callus of Hypericum perforatum L. reached 0.769 ‰ and 0.012 ‰dividually.The regeneration system of Hypericum perforatum L. stem, leaf and root were studied. Experiments showed that MS (1/2N) medium supplemented with TDZ 0.5 mg/L or 6-BA 0.5-1.0 mg/L was good for differentiation of stem; Shoot of leaf was easy to be induced on MS medium supplemented with 1.0-2.0 mg/L ZT; MS (1/2N) medium supplemented with TDZ 0.1 mg/L, or 6-BA 0.1 mg/L was suitable for differentiation of root. We formed a kind of stable plant regeneration system through the study.The Target gene Hyp was inserted into the corresponding position of the expression vector pBI121 and the plant expression vector which contained target Hyp gene was acquired. With the engineered Agrobacterium tumejaciens clones, transformation of the Hyp gene was conducted; Also transformation via particle bombing was conducted. 6 Km-resistant callus and 5 Km-resistant buds were acquired.