Transformation Of Tobacco With The HAL1 Gene Improving Salt Tolerance

Saline soil is very important agricultural production source.Greened saline soil is a key way for enlange agricultural areas, develop production, improve ecological environment.Using gene engineering to culture salt tolance plants is a efficient method for saline soil.HAL1 gene is a key factor of salt tolerance, it's expression can regulate Na⁺/K⁺ balance.Thought HAL1 gene is not a kind of transfererd protein,it can keep lower Na⁺/K⁺ with other gene and transfererd sistem.So HAL1 gene have great potentiality in the gene engineering of salt tolance.Based on the plasmid Prokâ…¡carried HAL1 gene was transferred into Agrobacterium tumefaciens LBA4404 and C₅₈C₁.HAL1 gene was transformed into tobacco by transfection of Agrobacterium tumefaciens,the factors effecting on frequency of gene transfer were examined and salt tolerance character of transgenic tobacco was analysised.The main results are as follows:1 . The plasmid Prokâ…¡carried HAL1 gene was transferred into Agrobacterium tumefaciens LBA4404 and C58C1.The results suggested that the best competent cells can be obtained when A.tumefaciens grew at OD600=0.5～0.6 with YEB medium,prepared with 70mmol/LCaCl2.LBA4404 competent cells mixed with 10ng plasmid DNA at 0℃for 30 min,then frozened by liquid N2 for 5min, followed heat shock at 37℃for 5min,the transformation frequency reached 1.40×10⁵transformants/μgDNA.C58C1 competent cells mixed with 20ng plasmind for 10min, frozened for 1min,heat shock at 37℃for 1min, the transformation frequency reached 1.33×105 transformants/μgDNA.2.Established the optimal tissue culture system by using tobacco as experiment material.The culture medium inducing bud from the leaf of"Qinxuan NO.1"and"Qinyan 95"were respectively MS + 6-BA2.0mg/L + NAA0.4mg/L, MS + 6-BA1.0 mg/L + NAA0.1mg/L. 20mg/L and 30mg/L Kanamycin were used to be selector for Kan-resistant shoots of"Qinxuan NO.1"and"Qinyan 95",20 mg/L Kanamycin was used to be selector for inducing roots.3.Factors were examined and an efficient gene transfer was established.40～60d regenerative leaves of"Qinxuan NO.1"were not pre-cultured and transferred with delusioned...