Introduction and Expression of Foreign Genes in Rice Cells by Particle Bombardment

For establishing a transformation system of rice, an efficient introduction of foreign genes into embryogenic cell suspension by particle bombardment was conducted. The particle inflow gun based on the acceleration of DNA-coated tungsten particles using pressurized helium was constructed for delivery of DNA into rice cells. Several bombardment parameters were optimized using the transient expression of GUS gene. The conditions that gave the highest GUS gene expression of about 1000 blue spots per g fresh weight of bombarded cells include treatment of the cells with 0.5 M osmotic pressure, and use of the 410 kPa helium, 110 mm target distance, 13 mm syringe filter holder and 5 $\mu$L DNA/tungsten mixtures. It was also confirmed that rice actin promoter-intron construct gave the highest expression of all promoter-sequences studied. Eight weeks after the bombardment, stably transformed calluses were obtained on the selection medium containing 100 mg/L G418 and showed the strong activity in in situ GUS assay.