| Lonicera macranthoides which was used as an excellent vertical plant and Lysimachia christinae which was used as a beautiful cover plant were chosen as materials. This two kinds of plants were not only used as landscape plants but also used as important medicinal plants. The complete rapid propagation system and regeneration system of L. macranthoides were estsblished. The rapid propagation system included primary culture, multiple shoot induction for proliferation, seedling culture, rooting and transplanting, which will provide a theoretical and experimental guidance for largescale industtial production in future. The L. christinae germplasm of our laboratory were used for germplasm enhancemant, including polyploid induction and study on the system of genetic transformation. The main results were as follows:1. Establishment of rapid propagation system of L. macranthoides. The explants of L. macranthoides were filtrated after immersion in 75% enthanol for 20-30s and then in 5.0% NaClO solution for 15 min. This was the optimal sterilization method.1/2MS+ 6-BA 1.0mg/L+IBA 0.1 mg was the best medium for induction of multiple shoots, with the proliferation multiple was 5.52.1/2MS+6-BA 0.1 mg/L+IBA O.lmg was the best medium for the strong seeding culture.1/2MS+6-BA 0.1 mg/L+IBA 0.2mg+AC 0.5 g/L was the best medium for rooting culture. The explant used for rooting should be semi-lignified which were chosen from seeding culture. The suitable illumination conditions were the former period with darkness culture for 5 days and the later period with light culture. After inoculated on the rooting medium for 5 days, the explants took roots and with the rooting rate 93.33% and the average rooting number 2.51. The best transplanting method with survival rate 94.44% were planted the train seedlings with roots in sand bed for 14 days and then transplanted them into humus:vermiculite:perlite with mixture medium soil by 6:2:2 (v/v).2. Establishment of regeneration system of L. macranthoides. Stem without sectong was the best explant for regeneration.1/2MS 6-BA 3.0 mg/L+IBA 0.1mg was the best solid medium for regeneration, with the regeneration rate 98.89% and the proliferation multiple 7.64.1/2MS 6-BA 4.5 mg/L+IBA 0.15mg was the best liquid medium for regeneration when the stem without sectong was the best explant, with the regeneration rate 100% and the proliferation multiple 11.82.3. Tetraploid of L. christinae was produced for the first time. Survival rate decreased when the concentration of Oryzalin and incubation time increased. Stem with section incubated by Oryzalin had a lower survival rate than stem with section and half of a leaf incubated by Oryzalin.10?mol/L Oryzalin incubated 48h had the highest mutational rate of 61.11%. After chromosome doubling, and tested by the folw cytometry, tetraploids of L. christinae were produced.4. Induction of callus of L. christinae. Leaf and stem without section of L. christinae were chosen as the explant for the induction of callus experiment. MS+2,4-D3.0 mg/L+6-BA 0.4 mg/L was the best medium and stem without section was the best explant for the callus induction. MS+6-BA 0.1 mg/L+NAA0.1 mg/L was the best medium for the regeneration of callus. There were significant differences between diploids and tetraploids in morphology such as diameter, internode length, leaf shape, stomatal flower patterns.5. In the study of genetic transformation system, medium with Cef 300 mg/L not only can effectively antimicrobial but had the mimium inhibitory effect on explant regeneration. Screening antibiotics was Km 50mg/L. The system of genetic transformat-ion still needs further study. |
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