Tissue Culture And Studies On The Secretory Structure And Their Relations To Hypericin Content In Hypericum Perforatum And Hypericum Erectum

Hypericum perforatum and Hypericum erectum,wild perennial, belong to the genus Hypericum from the family Guttifeae.They have important medicinal value.A great deal of attention has been paid to the genus because of the two main active hypricins and flavonids which are used for the treatment of mild to moderate depression,antivirus, anticaccer and improve immunity.In the present study,the two plants were investigated in tissue culture,phytochemistry,anatomical, relationship between the secretory structure and hypericin accumulation points of view.These researches provides a significant scientific basis for H.perforatum and H.erectum.The main results obtained from the experiment were summarized as follows:1.The inductions of callus from stem,leaf were systematically studied in H.perforatum and a great deal of callus were obtained. Meanwhile,quantity of hypericin was analyzed.The experiment showed that the calli come from stem formed easier than from leaf.The optimal medium for callus indction was the MS medium supplimented with 2.0 mg/L 6-BA and 0.2 mg/L NAA.And the callus cultured on the MS medium supplimented with 0.3 mg/L 6-BA and 1 mg/L 2,4-D growed rapidly.HPLC analysis of hypericins proved that callus from leaves showed higher hypericins levels(0.0025%)than that from stem(0.0021 %).2.The tissue culture and plant regeneration of H.erectum were systematically studied and a large number of regenerated plantlets were obtained.The results were as follows:Callus was inducd from the sterile plants from seed and the optimal medium was the MS medium supplimented with 0.2 mg/L 6-BA and 1 mg/L 2,4-D.The highest value of shoot multiplication(7-fold)from sterile germinated seedlings was obtained from MS medium in the presence of 0.2 mg/L NAA and 2.0 mg/L 6-BA.Roots developed on the 1/2 MS medium were long and numurous.3.The relationship between the development of black nodules and hypericin content was also analysed in H.perforatum and H.erectum. And the result showed that biosynthesis of hypericins is connected with the formation of black nodules.No hypericin was observed in the undifferentiated callus which was consistent with the HPLC result.4.Hypericins were found in shoots,leaves,plantlets without roots and the content of hypericins generally increased during their development.HPLC analysis of hypericins proved that leaves from plantlets 5 cm high showed higher hypericins levels(0.00554%)than other regenerated organs and plantlets in vitro.These results showed that H.erectum in vitro cultures represent promising systems for hypericin and pseudohypericin production.5.We studied the content of hypericins in a multicellular nodules and a secretory cell of H.perforatum and H.erectum for the first time. And the result showed that,63.12%and 55.50%of total hypericins were quantified in a secretory cell of H.erectum and H.perforatum respectively,while,only 29.88%and 30.04%of hypericin were found in a secretory cell.