Studys On Transformation Of Glucose Oxidase Gene Into Rice And Its Expression

Glucose oxidase (GO) could oxidize glucose to generate hydrogen peroxide (H2O2), H2O2 not only have a direct killing effect for pathogenic bacteria, but also can be used as a signaling molecule to activate a series of plant defense gene expression, so that plants produce a systemic acquired resistance. Studies also found that oxide burst in incompatibility of plant-pathogen interaction resulted in the produce of hydrogen peroxide, which not only as a regional signal could lead to cell death, but also regarded as a proliferated signal inducing defense genes expression in surrounding non-infected cells, such as glutathione -S-transferase gene.In this study, glucose oxidase gene was inserted into pCAMBIA3301, the efficient plant expression vector was constructed, with Ubiqutin-promotor and bar gene as selectable maker. The plasmid was transmitted into Agrobacterium tumefaciens EHA105, mature embryogenesis callus of Zhonghua11 variety was used as explant with Agrobacterium-mediated method, and the callus selected by Basta would differentiate into regenerated plant. The polymerase chain reaction and Basta spraying leaves were used to test whether the GO gene had been integrated into rice genome, and starch-KI staining and titanium superoxide method was used to measure contents of H2O2 in transgenic rice, the result indicated that glucose oxidase resulting from the GO gene had functioned in the rice. The activity of antioxidant enzymes also showed that the expression of GO gene increased the activity of the enzymes. As the hydrogen peroxide-induced plant defense response was not species-specific, therefore, transgenic plants would be resistant to a wide range of pathogens.The pUBGO Vector with bar gene as selectable marker, provided a great convenience for field screening and hybridization which could transfer the GO gene into other rice varieties.