| Bolbostemma Paniculatum F. belongs to Bolbostemma Franquet, Cucurbitaceae. Tubrimoside is a kind of effective component in it and is active to virus and cancer. Hypericum perforatum L.belongs to Euhypericum, Clusiaceae, and hypericin is a kind of effective component in it and is active to virus and cancer, too. A systematic study on the tissue culture, separation and extraction of effective components from Bolbostemma Paniculatum F. and Hypericum perforatum L. has significance not only in enriching the theory of plant tissue culture ,but also in breeding their new cultivars and producing their active components by biotechnology. This paper using Bolbostemma Paniculatum F. and Hypericum perforatum L.as materials, established and perfected their tissue culture and plant regeneration systems, studied the effective component separation and extraction processes. The main results were as follows:1. The induction of callus from stem, leaf were systematically studied in Bolbostemma Paniculatum F. and a great deal of calli were obtained. And the quantity of Tubeimoside was analyzed. The experiment showed that the calli come from stem formed earlier than from leaf. The stems cultured on the MS medium supplemented with different kinds and concentrations of hormones could form a lot of calli. The leaves cultured on the MS medium supplemented with 2,4-D?2,4-DandNAA, NAA and BA could form calli, The growth rate of callus on the MS medium was higher than on the 85 medium, but it was easy to become brown.The calli cultured on the MS medium supplemented with 2,4-D2.0 mg/L, NAA0.5 mg/L* BA1.0 mg/L were optimal for Tubeimoside I production. The calli cultured repeatedly on the 85 medium supplemented with 2,4-D2.0 mg/L* NAA0.5 mg/L* BA1.0 mg/L could form a kind of even stable and callus line.2. The tissue culture and plant regeneration of Bolbostemma Paniculatum F. stem and leaf were systematically studied and a large number of regenerated plantlets were obtained. The experiment showed that numerous shoots could formed directly when leaf explants were cultured on the MS medium supplemented with BA3.0 mg/L * NAA 1.0 mg/L and calli were cultured on the MS medium supplemented with BA2.0 mg/L and IAA1.0 mg/L. The MS medium supplemented with BA2.0 mg/L,IAA0.13mg/L was optimal for shoots growth .Detach the shoots and transfer them to 1/2MS medium supplemented with 1.0 mg/L IBA for rooting and find that 50% shoots produced roots after 15 days culture. At pH6.0 and 0.8-1.0% agar was optimal for callus and shoot formation, while pH5.8 and 0.6-0.7% agar was optimal for root formation.3. The tissue culture and plant regeneration of Hypericum perforation L.stem and leaf were systematically studied and a large number of regenerated plantlets were obtained.The MS medium supplemented with BA0.2 mg/L,2,4-D(3.0-4.0mg/L) was suitable for the induction of callus; The MS medium supplemented with BA(3.0-5.0mg/L),NAA(0.5-1.0mg/L) was good for the induction of shoots; The MS medium supplemented with BA0.05 mg/L,NAA1.0 mg/L or IBA1.0 mg/L was good for the rapid propagation of shoot; The MS medium supplemented with BA1.0 mg/L or 1/2MS medium was good for rooting. The experiment showed that numerous shoots and plantlets could be produced in a short time from the calli, which formed from stem, leaf explants cultured on the MS medium supplemented with higher concentration radio of BA/NAA. But meantime, many hyperhydric shoots were produced. A critical technique for the rapid propagation of Hyperium perforatum L.is that the concentration of BA is higher in the early stage of culture and then lower in the later stage. By this method, a numerous normal plantlets could be obtained and the number of hyperhydric shoots was few.4. The stems and leaves of Bolbostemma Paniculatum F. and Hypericum perforatum L. were cultured and observed. Two kinds of calli formed in all explants. One was hard, while the other was fragile. Both of calli come from Hypericum perforatum L.could produce shoots and the shoots formation frequen... |
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