A Highly Efficient Transformation System Mediated by Agrobacterium tumefaciens in Rice(Oryza sativa L.)

Abstract

After the factors influencing transformation were carefully investigated, a highly efficient Agrobacterium-mediated gene transfer system has been developed for rice. Using this protocol, inunature embrys of rice variety Zhongha No. 11 cultured for 4 days were infected and haformd by A. tumefaciens stain EHA105 harboring Ti Plasmid pCAMBIA1301 The results showed that infected embryos gave a high hequency (50% ～ 90%) of GUS gene expression after co-cultivation. The hansformation efficiency of stable integIation of TDNA reached 87. 6% measured as hypomycin-resistan calli produced and 64. 6% measmed as numberof hansemc plants regenerated, respectively. Southem blotting analsis of the total DNA extrated frOm transgenic plants indicated tha the TDNA had been integrated into the rice genom, and most of bosgenic planis had one coPy of TDNA insert. The results of genetic analsis ford tha the bosgenes were segngaed in a Mendelian fashinonin the T1 generation.The establishment of this bosformtion system in rice Provides an efficient way for transferring usend foreign genes to rice plants