Establishment of an Efficient Genetic Transformation System for Rice with Glyphosate as a Selection Marker

PurposeTo establish an efficient rice genetic transformation system with Nipponbare as the receptor and glyphosate resistance gene (CP4) as the selection marker for transformation research.

MethodUsing mature embryos of japonica rice Nipponbare as experimental materials, the P1300 vector containing the glyphosate resistance gene CP4 was transformed into the callus of Nipponbare by Agrobacterium-mediated method, with a concentration of 10, 20 and 40 mg/L glyphosate were subjected to three rounds of resistance screening, differentiation induction and seedling formation. l 000 mg/L glyphosate was used to identify the resistance of the positive seedlings transplanted alive.

Result145 transgenic seedlings were obtained. After PCR detection, 128 positive strains were obtained, the positive rate was 88.27%. Experiments proved that after three concentration gradients screening, the positive rate of genetically modified seedlings increases. Among the 99 positive seedlings transplanted alive, 65 strains showed resistance, and the resistance rate was 65.66%.

ConclusionWe successfully established a rice transgenic system using glyphosate as a screening marker. The system provided materials for rice breeding and provided technical support for rice transgenes using glyphosate as a screening marker and functional verification of some genes.