Construction Of SeNHXⅠExpression Vector And Transformation Into Campanula Medium

High-concentration sodium ions can cause salt stress, resulting in deleterious effect on critical biochemical process and tissue injury, including ion toxicity, osmotic stress, and Na~+/H~+ ratio imbalance. The Na~+/H~+ antiporter transports Na~+ across the plasma membrane and vacuolar compartmentation, reduce the accumulation concentrations of sodium ions, at last the resistance to salt stress improves plant salt tolerance. The molecular genetic manipulation on Na~+/H~+ antiporter gene leads to a better understanding of its function, which promotes the development of gene engineering in plant salt tolerance.SeNHX1 belongs to Na~+/H~+ antiporter family. We constructed the plant expression vector pBin121-SeNHX1 using the binary expression vector pBin121 with 35S promoter region, kanamycin resistance gene and GFP (green fluorescent protein) gene. Then, pBin121-SeNHX1 was transferred into the agrobacterium C58.The second aspect of our experiment is about the establishment of Campanula medium regeneration system. Tissue culture of Campanula medium is firstly established in our work. Through 2, 4-D, 6-BA, NAA different combinations of hormones, we screened out the callus induction medium, bud differentiation medium, rooting medium and eventually established Campanula medium tissue regeneration and genetic transformation system, which laid the foundation of the further Agrobacterium infection.At last, by the use of Agrobacterium-mediated transformation methods, SeNHX1 gene was introduced to the target plant. Through initial detection to resistant seedling and wild-type plant control, including fluorescence detection, PCR method, and SeNHX1 gene was successfully transferred to Campanula medium.