Cloning,Expression And Function Analysis Of Soybean AKT1 Gene

Soybeans are the most widely grown oil crops in the world and one of the most important food crops in China.However,soil quality in China is generally low,and in some areas,droughts,natural disasters such as salinity and alkalinity can seriously affect the yield or even the harvest of soybeans.If these lands can be used by some crops,our country's grain will not have to rely on foreign imports.Therefore,the present study analyzed the expression of soybean GmAKT1 under different stress conditions.The plant expression vector pBASTA-GmAKT1 was constructed and the target gene was transformed into wild-type Arabidopsis thaliana by floral leaching method.Subcellular localization studies and soybean hairy root phenotype studies were conducted to lay the foundation for further exploration of the function of the GmAKT1 gene.The main findings are as follows: 1.The expression of GmAKT1 gene in soybean roots and leaves under different stress conditions was analyzed by real-time fluorescence quantitative PCR.The results showed that the expression level of GmAKT1 gene was significantly increased at the initial stage of salt stress stress,and the expression amount Was controlled in leaves 3.65 times.Under salt stress and saline-alkaline stress conditions,the expression of GmAKT1 gene in roots was significantly increased,and the relative expression level was 2.15-fold and 2.35-fold higher than that of the control group.There,with the increase of stress time,the expression level of GmAKT1 gene gradually decreased in both root and leaves.There was no significant change in the expression level between the alkali and drought stress and the control group.2.GmAKT1 was constructed by gene engineering technology into the pBasta plant overexpression vector,and GmAKT1 was transferred into the wild type Arabidopsis thaliana by Florial Dip.A total of 9 T1 transgenic plants were obtained.3.By identifying the T2 transgenic plants of Arabidopsis thaliana,9 strains showed that the segregation ratio was 3:1,which accorded with the Mendelian law of inheritance,and the three lines with the highest expression levels were identified for further gene function.The identified studies provide plant material.4.The GmAKT1 gene was constructed by DNA recombination technology into pCAMBIA1302 vector.GmAKT1 was transferred into tobacco mesophyll cells by Agrobacterium injection method.It was confirmed by laser confocal microscopy that GmAKT1 was localized on the cell membrane.5.A soybean hairy root induction culture system was established to induce GmAKT1 soybean hairy roots.The gene expression of GmAKT1 soybean hairy root under salt stress was analyzed.Phenotypic analysis of GmAKT1 soybean hairy roots was performed.It observes changes in stomata under salt stress.The results showed that the hairy root expression level of GmAKT1 was significantly higher than that of the control group.Under adverse conditions,the closure rate of stomata was higher than that of the control group.