Floral Dipping System Optimization Of Transferring Populus Euphratica Large Fragment Gene To Arabidopsis Thaliana And Mutants Screening

In this research, tall trees Populus euphratica and model plant Arabidopsis thaliana were used as research material in this transgenic researchs plan. The large fragment genes from Populus euphratica were transferred to Arabidopsis thaliana by floral-dip method mediated by Agrobacterium tumefaciens. The surfactants SilwetL-77were used in different concentrations and dipping time to transfer large fragment genes of Populus euphratica carrying by Agrobacterium into Arabidopsis thaliana. The obtained seeds were screened from resistant culture media, then possible Arabidopsis thaliana transformants were checked by DNA extract, PCR test and phenotypic changes screening. In order to further study on Populus euphratica genomics we need to obtain the homozygous Arabidopsis mutants which carry large fragment genes of Populus euphratica and those genes could be stably inherited, then positive transformants would be continued to culture and make resistance screening and passage genetic differences screening.The main results were as follows:(1) The superior conditions for activating Agrobacterium were YEP liquid medium using and24hours culturing. The superior transformation system was as following,300ul/L SilwetL-77concentration in infection solution and infecting for7min,500ul/L Silwet L-77concentration in infection solution and infecting for lmin.(2) Totally screened and extracted1500Arabidopsis transformants DNA from the862clones, most of which exhibit positive, obtained171positive transformants. The better disinfection method is alcohol applying disinfection which is used in Arabidopsis mutants screening.(3) Totally obtained7mutants which have phenotypic changes after screened repeatedly and offspring observed.(4) Some changes had be made on the cultivation of Arabidopsis thaliana to let them grow strongly under the specific environmental conditions and conducive to transforming.This thesis may make a great contribution to complete the gene banks of Populus euphratica as well as provide the basis for the study on the role of Populus euphratica genome...