Cloning And Transgenic Research Of ?-glutamate Cysteine Ligase Gene In Potato

As one of the important staple crops,potato is affected by cadmium stress during its growth and the development.In plant cells,glutathione as one of the determinant regulators in cadmium stress response.In this thesis,potato ?-glutamate cysteine ligase gene(StGCL)was studied in order to clarify the molecular mechanism of StGCL affected glutathione synthesis in potato and provide a new way for further heavy metal stress-resistant potato breeding.The main research results are as follows:1.StGCL gene was cloned.It has high homology with GCL genes in Solanaceae(tomato,pepper and tobacco).Amino acid sequences analysis showed StGCL has two cysteines which form the core disulfide bridge(CC2);has amino acid sites where the GCL is responsible for substrate binding and catalysis;may forms protein dimers.2.StGCL overexpression and RNAi-related genetic transformation were performed.StGCL overexpression strains were identified,result showed that the GSH content in the overexpressing strains were increased significantly.3.Subcellular localization analysis of StGCL showed that it localized to the nucleus and the cell wall.