| In this paper, mature spores and young sporophyte of Osmunda cinnamomea L.var.asiatica were chosen as explant to study Propagation and production in vitro. Discussing different medium, hormones, culture conditions effect on spore germination, subcultureing and proliferation of prothallus and sporophyte transforming.Summed up a method of rapid propagation young sporophyte of Osmunda cinnamomea L.var.asiatica. Meanwhile, Two young sporophyte of Osmunda cinnamomea L.var.asiatica were used as explant to research corresponding dedifferentiation in order to built perfect propagation system of Osmunda cinnamomea L.var.asiatica in vitro culture and settle base for transgenic seedling.The results were as follows:(1) It is esay for spore germinate to reduce inorganic salt concentration.the spores from broken sporangia inoculated in medium of 1/6MS with peptone0.5g/L+sucrose20g/L+agar8.6g/L and 1/4N6 with peptone0.5g/L+sucrose20g/L+agar8.6g/L were easy to germinate quickly in the condition of changeable temperature and light and rate of germination is bove 97%, spore germinate after 4 days and bevisible between 12-15 days. Effect useing sugar instead of sucrose is not difference on spore germination.(2) It is esay for subculture and proliferation of prothallus to reduce inorganic salt concentration and which of the best is 1/2MS or 1/2N6.1/2MS culture medium added with 8g/L liquid extracted from leaves of Osmunda cinnamomea L.var.asiatica or KT5.0mg/L, NAA0.5mg/L and NAA1.0mg/L, both of which were good for promoting proliferation of prothallium. Temperature of subculture and proliferation is 25℃.(3)The spore which were inoculated in culture bottle shaped young sporophyte that totaled about260 days. while the sporophyte induction which transformed outside cultivating bottle totaled about 240 days. The time of selling seedling need one years and eight months. Induction outside bottle were suitable for rapid propagation young sporophyte of Osmunda cinnamomea L.var. The main research were as follows:①Inoculating 60～80 pieces of prothallus in culture bottle with 9cm bottom diameter, after 120 days the maximum conversion rate of sporophyte was 31%, which reached the maximum value after 180 days, and it was defined that the changing of temperature and light was benefit of sporophyte's conversion and growth.The duration time of sporophyte's subculture is 90 days, and after 4 duration times the growth of subculture was improved obviously, such as its rhizoid became clearly, breeding rate became slowly, and the conversion rate of sporophyte became lower. Chosen the seedings with thick roots, height under 5cm, leaves more than 3 and roots more than 6, to culture on Osmunda cinnamomea L.var original soil, the survival rate was 81% at last.②Transplant the prothallus of 2 monthes breeding on the Osmunda cinnamomea L.var original soil, with 2000-3500Lx scaterring light intesity,26 C room temperature,above 90% air humidity,and potting induction medium once a week, the induction rate of sporophyte were above 53% after 60 days, the average height of seeding was 5.4 cm and roots were more than 5 after 180 days, the sur-vival rate was above 96% when divise transplanted.(4)Study the regeneration ability of 2 types of Osmunda cinnamomea L.var explants primarily,the result was: the best sterilization time by 0.1%HgCl2 was 3-4 minutes about mature sporophyte leaves and 10 minutes about root stem terminal bud, and the meduim was 1/2MS+KT1.0mg/L+2,4-D1.0mg/L +NAA1.0mg/L+sucrose30g/L; the action of dedifferentiation could be made by 1/2MS+BA1.0mg/L+N AA1.0mg/L,but the callus of leaves were very hard to dedifferentiate buds, lots of them were dediff-erentiated to roots, so the callus came back to prothallus stage. The callus of root-stem terminal buds dedifferentiated could dedifferentiate adventive buds and then generated complete sporophyte plants in the medium of 1/2MS+6BA0.1mg/L+NAA2.0mg/L+ZT1.0mg/L+VC2.0mg/L and MS+KT1.0mg/L+NAA 0.1mg/L+VC2.0mg/L. |
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