Transformation Of Pac 1 Gene Into Tobacco Via Agrobacterium-mediated Method

In this paper, tobacco seedling leaves were used as explants and pac1 gene were transferred into important tobacco cultivars Longjiang911-21, Jiyan9 and Yunyan87 by Agrobacterium-mediated method for the first time to obtain resistance to tobacco viruses.Kanamycin resistance plants, were obtained, that were 43 plants of Jiyan9, 61 plants of Longjiang911-21 and 43 plants of Yunyan87.Genomic DNA of the kanamycin positive plants,which extracted with CTAB method were used templates for PCR detection.The results showed that positive ratio of Yunyan9 was 21 to 43,that of Longjiang911-21 was 32 to 61 and that of Yun yan87 was 24 to 43.Specific corresponding fragment were amplified for all of positive tobaccos by PCR detection, which identified that pad gene has been transferred into tobaccos.In order to further determine integration of pac1gene in the transgenic plants, four tobaccos selected randomly from transgenic plants were used for Southern Blot analysis with non transgenic tobaccos as control,the results indicated that pac1gene has been integrated into genome DNA of tobacco successfully.