Development Of Regeneration Systems Of Four Chinese Herbs And Preliminary Determination PPG In Pedicularis Kansuensis Maxim

The tissue culture and regeneration systems of Pedicularis kansuensis Maxim, Arctium lappa L., Vaccaria segetalls, Potentilla potaninii Wolf which were Chinese medical were studied in this thesis. The content of PPG (Phenylpropanoid Glycosides) in Pedicularis kansuensis Maxim was determined with TLC (Thin-layer chromatography).1. In vitro regeneration system of Pedicularis kansuensis Maxim had been developed firstly to date. The germination rate of seeds was improved by dipping in water at low temperature and dealing with hormones. The fragments from cotyledons, hypocotyls of Pedicularis kansuensis Maxim as explants cultured on MS medium supplemented with different concentrations of hormones. It showed that the root was the best explant to induce callus and the callus from hypocotyls formed shoots easily. Histological studies showed that the embryogenic cells assembled in the surface or inside of callus. The elongated shoots were detached and transferred to medium that be used to induced root. The frequency of rooting was about 60%. About 41.6% of the plants were successfully established in soil after one month. Pedicularis kansuensis Maxim callus contain much glucose and saccharose, and there are a little PPG in the regenerated plants.2. An efficient and reproducible procedure was established for the plant regeneration from cotyledons, hypocotyls, and cotyledon- or hypocotyl-derived calli of Arctium lappa L. The combinations and concentrations of different growth regulators were shown to be critical factors for the frequency of callus induction as well as for the potential of callus differentiation. Callus was induced from cotyledon and hypocotyl explants of Arctium lappa with 87% ~ 100% frequency in MS medium supplemented with 2.0 mg/L 2,4-D and 0.5 ~ 2.0 mg/L BA. The shoots were induced indirectly from calli or directly from explants of Arctium lappa on MS with 1.0 ~ 3.0 mg/L NAA and 0.5 ~ 2.0 mg/L BA. The shoot forming rate from hypocotyl was obviously higher than that from cotyledon, and the direct differentiating frequency inhypocotyls was 77.3% on MS with 1.0 mg/L NAA and 1.0 mg/L BA. Histological study showed that the shoots regenerated via organogenesis or somatic embryogenesis in Arctium lappa. The shoots were rooted on 1/2MS with 1.0 mg /LIBA and 1.0 mg/L NAA, and about 93.3% of the regenerated plants were successfully transplanted to soil. The whole procedure needed about 13 weeks. The regenerated plants of Arctium lappa flowered and set seeds next year, with normal morphology and growth characters.3. The tissue culture system with high frequencies of regeneration had been developed in Vaccaria segetalls. The calluses were induced from cotyledons, hypocotyls of Vaccaria segetalls. Then the differentiation of shoots was on the low concentration of NAA (O.lmg/L and 0.2mg/L) and these adventitious shoots rooted on half MS medium very easily.4. A protocol of high frequency organogenesis and plant establishment has been developed for Potentilla potaninii Wolf. Hypocotyl, cotyledon explants of Potentilla potaninii were cultured on MS media supplemented with various concentrations of BA and NAA to induce adventitious shoot regeneration and micropropagation. The highest frequency of adventitious shoot regeneration was achieved from hypocotyl and cotyledon explants on MS media supplemented with 5.0 mg/1 BA and 1.0 mg/1 NAA. The development shoots rooted best on half-strength MS medium supplemented with 1.0 mg/1 NAA and 0.5 mg/1 IAA or IBA. The regenerated plants of Potentilla potaninii flowered and set seeds next year, with normal morphology and growth characters.Establishment of these medical herbs in vitro regeneration systems is not only an important step that will open new avenues for further researches, but also useful for the maintenance of entironment.