Establishment Of Regeneration System And Preliminary Study Of Genetic Transformation By Aarobacterium Tumefaciens Of Sudangrass

Sudangrass is one of the important species of sorghum, which is primarily used as feed and fodder for animals. Sudangrass is a C4plant that is characterized by a high photosynthetic efficiency, which can yield a high biomass. It has been reported that the tolerance of sudangrass to drought and high temperature is higher than that of corn, wheat and other crops and sudangrass has the unique ability to grow under a wide array of harsh environmental conditions. This experiment studied the effect of the inducing of callus, regeneration and rooting, and primary callus infestation were found, providing a basis for gene transformation.In trial1young inflorescence were applied as explant material and MS base culture were used in all experiments. The induce rate were appraisaled by the concentration of hormone, AC, CH and Pro with orthogonal design. The regeneration rate were appraisaled by the concentration of the type of the medium, the concentration of hormone, AgNO3and CuSO4. The root rate were reviewed by the concentration of the type of the medium, the concentration of hormone. The results were listed as follows:(1) The best and inferior effect of the callus induction were as follow:2,4-D>KT>CH>Pro>AC. Callus induction and the best statues callus were got at the concentration2,4-D was4mg/L, KT was0.1mg/L, Pro was2000mg/L, AC was50mg/L, CH was1000mg/L.(2) The highest regeneration rate was50%at the concentration of0.1mg/L NAA and2mg/L6-BA, and the regeneration rate was improved from50%to69.57%by addition into0.5mg/L CuSO4. Since the highest regeneration rate was41.12%at the concentration of0-10mg/L AgNO3, so it may not some effects on the callus regeneration of sudangrass.(3) The highest rooting rate was97%at the concentration of0.1mg/L NAA and the cultural was1/2MS.In trial2different type callus were used in conforming the optimal callus which infestation by Agrobacterium tumefaciens-mediated with the paraffin section analysing, the content of the DNA and the regeneration rate. Agrobacterium strain, OD600value, AS concentration, AC concentration and the time of co-culture were ascertained by the one-factor analysis of variance, Transgenic resistant hph callus are detected by GUS detecting. The results told that the type Ⅲ was the best callus type when compared with others. Genetic transformation has been established. By Agrobacterium-tumefaciens mediated, using the Agrobacterium strain LBA1301, OD value was1.2, AS concentration was20mg/L, AC concentration was50mg/L, the time of co-culture was3d. genetic transformation was optimized.