Establishment Of Genetic Transformation System Of Sedum Spectabile Boreau By Agrobacterium Rhizogenes

The infancy stem segments Sedum Spectabile Boreau of were taken as explants to be cultured, through which their floral system was established in this experiment. Besides, callus of the leaves and stem segments and the infancy stem with shoot segments of Sedum Spectabile Boreau were cultured in the different medium of concentrations of hormone. In order to provide genetically improved new germplasm resources and achieve the transgenic plants of Sedum Spectabile Boreau ,genetic transformation system was established based on the system of regeneration. Several factors affected genetic transformation of Sedum Spectabile Boreau mediated by agrobacterium rhizogenes were studied. The shape characteristic, PCR detection of transgenic plants were researched, shape and Growth variation and leaf anatomic structure were also studied. The main research work and results were as follows:1.Regeneration system for Sedum Spectabile Boreau leaf explants was established,the experiment results he resulte showed that good propagation medium was MS0,good rooting medium was 1/2MS0 , the good medium of leaves callus induced was MS+6-BA 0.3mg. L-1+NAA 0.1mg. L-1. Based on this suitable medium, the effect of dark cultivation on induction of adventitious buds from leaves explant was studied. It was found that dark cultivation for 5 days would be favorable for induction of adventitious buds.2. Effects of Cefotaxime (CTX) on induction of adventitious buds from Sedum Spectabile Boreau of triploid P. tomentosa explants and inhibition of Agrobacteria were investigated. The results indicated that 400mg.L-1 was a suitable concentration to control the propagation of Agrobacteria.3.Several crucial factors influencing the transformation efficiency were studied. The results showed that: the times of activation, the time of infection, the concentration of agrobacterium rhizogenes's solution esc, had some effect on transformation frequency. The highest transformation frequency were infected for 10 min with the concentration of agrobacterium rhizogenes's solution OD600=0.4,and then co-cultivated for 2d .4. PCR identification method was used on transplant, the results indicate T-DNA of Ri plasmid had already been combined into the transplant. 5.Shape and growth variation of transgenic plantlets were also studied.The result indicated that most of transgenic plantlets with Agrobacteria exhibited dwarf plant ,increased leaf area,increased branch, shortened internodes,decreased rooting rate ,root number and growth of stem length .