Establishment Of The Regeneration System From Leaves Of Ruby Seedless And Rootstock SO4

The first transgene plant has been obtained in the world since 1983, as the improvement of transgenic technic.Transgene technology has been developed as a kind of most important instrument of plant breeding, it provided a new way for breeding grape. Now, too low regeneration rate restricted transgene project developing of grape. Although there were many reports in grape tissue culture, most of them were the culture of bud axillary explants, these is not fit to transgene regeneration. Therefore, the mainly important of study aftertime were searching a high efficiency regeneration way and establishing a steady system of grape's decend and transform.In this study, "Ruby Seedless" and "Rootstock SO4" were used as the experiment materials. After sterilize,initial culture,subculture and multiplication, leaves of grapes in vitro were used as explants, studying the effect factors on regeneration, such as the different hormone combinations, the different inoculated fashion and so on. The leaf regeneration system had been successful established. The main results were as followed1 To sterilize single bud stem of grape with 0.1% HgCl solution,and results showed that the 0.1% HgCl sterilize 13 min has the best effect for sprout.2 Subculture and Multiplication:The best subculture medium of ruby seedless was MS+6-BA0.8mg/L+IBA0.2 mg/L. the best culture medium of SO4 was MS+6-BA1.0 mg/L+IBA0.2 mg/L.3 Leaf regeneration system:the leaf best hormone match of Ruby seedless was wasMS+TDZ2.5mg/L+IAA0.05mg/L, regeneration rate was 23.1%; rootstock SO4 was MS+6-BA 4.0mg/L+IBA0.10 mg/L, regeneration rate was 50.9%; the top 2 leaves is the best explants for regeneration; the inoculated fashion of leaf abaxial to medium and dark incubation(21d) could improve regeneration rate.4 Most of regeneration bud can take root. The best rhizogenesis was observed on 1/2MS+IBA0.4mg/L+0.1% active carbon. The taproot strong and distributes radially,it is the ideal result for rhizogenesis. The rooting tissue culture seedling in unsealed tube 6days can improved the survival rat of transplanting.