Studies On Micropropagation And Plant Regeneration In Vitro Of Tengcha(Ampelopsis Grossedentata)

Tengcha （Ampelopsis grossedentata）, a kind of wild liane, belongs to Ampelopsis of Vitaceae. In recent years, the germplasm resources of Tengcha have been destroyed seriously because the villagers excessively picked and excavated the feral Tengcha for its high value in medicine and nutrition. Therefore, the study on the genetic diversity and conservation of Tengcha has the profound significance for preserved the germplasm resources, used rationality and developed economy. The extremely urgent research of Tengcha are the germplasm resources effective management, introduction, domesticates, cultivation, breeding improvement. The integration of the traditional method and modern breeding technique, especially application modern biological technique, will reduce the breeding cycle and enhance the breeding efficiency. The material of Tengcha collected from Laifeng county of Enshi municipality, Hubei province. The studies Established fast, highly effective and economy micropropagation system and plant regeneration in Vitro of Tengcha. The main research results as follows:1. During the annual sterilization of explants, using the stem segments of Tengcha as the explants for initial culture, the best season for collecting samples was March, in which germination rate was highly with less pollution. The optimal method was 75%ethanol 20s+0. 1 % HgCl₂8min+Tween-80 1～2 drop.2. The optimal medium for axillaries bud induction was B5+BA 1.0 mg/L+NAA 0.05 mg/L.3. During subculture, if using BA+NAA, the optimal multiplication medium was MS/B5+BA1.5 mg/L+NAA0.05 mg/L. The optimal strong seedling medium was MS/B5+BA1.0 mg/L+NAA0.05 mg/L. if using BA+Ade, the optimal multiplication medium was MS/B5+BA 0.5 mg/L+Ade2.0mg/L. The seeding in BA+Ade grew better and more than in BA+NAA.4. Between the two different auxins tested, NAA was the more efficient than IBA. However, shoots cultured on the medium which contained higher NAA formed too much callus on the bases to facilitate shoot growth and plantlet survival so that 1/2MS+IBA0.05～0.5mg/L was the best medium for rooting.5. The acclimated method of seedling was to open middle bottleneck for 2～3 d and then open all bottleneck 3d in the light, add to distilled water with the temperature about 25℃. The seedlings were transplanted in the mixture of soil, vermiculite and perlite（1:1: 1）.6. The best explants for callus induction was stem segments and the best media for callus induction was B5+BA 0.5mg/L+2,4-D 0.5 mg/L. The suitable media for callus subculture was B5+BA0.5～1.5mg/L+NAA0.01～0.1 mg/L. Light condition has predominant effect on the state of callus and the best time of callus induction.