한국 재배종 낙동벼에서 임성 형질전환식물체의 재분화

Abstract

Rice is one of the most successful monocots in regenerating fertile and genetically stable transgenic plants. However, there is no report of a rice line developed in Korea that can be used for regeneration of fertile and genetically stable transformants. In this paper, we first demonstrate that a Korean variety, Nakdongbyeo, is suitable to obtain transgenic rice plants. Protoplasts from embryogenic suspension cultures were co-transformed with HPT (hygromycin phosphohansferase) and GUS (β-glucuronidase) genes in seperate plasmids in the presence of PEG (polyethylene glycol). In 5 independent experiments, the average frequency of calli showing hygromycin resistance were 1.73%. Plantlets were regenerated from the Hyg^R calli. The average efficiency of plantlet regeneration was approximately 27%. Based on the GUS activities of hygromycin resistant calli, ca. 35% of the resistant calli carried active GUS genes. The R0 transgenic plantlets were grown to maturity and R1 seeds were obtained. By examining the in situ activity of GUS in R1 seeds and seedlings, we confirmed that the GUS transgene driven by a CaMV 35S (cauliflower mosaic virus) promoter, showed proper expression patterns. We also confirmed Mendelian segregation of the HPT transgene in the R1 generation.