Agrobacterium-mediated transformation of Nang Thom Cho Dao, an indica rice variety

Abstract

Agrobacterium-mediated transformation is the most efficient method to transfer gene of interest into the plant genome. However, the transformation efficiency of this method with indica rice varieties is still low and needs to be improved. In this study, the Agrobacterium-mediated method was used to transfer reporter genes into the Nang Thom Cho Dao, an indica rice variety. Different transformation parameters, such as selection to used hygromycin at concentrations of 0, 25, 30, 40, and 50 mg/L, bacterial density, and infected duration, were tested and optimized. The results showed that 30 mg/L of hygromycin was the most appropriate concentration for selecting transgenic callus in Nang Thom Cho Dao. In addition, the highest transformation efficiency of this Nang Thom Cho Dao rice cultivar was observed at the bacterial suspension density OD600nm of 0.1, the infected duration of 20 minutes, and the 3-day co-culture period. The presence and expression of transgenes were confirmed by gus staining and PCR with specific primers for hptII and gus genes. This transformation procedure should be used for further studies in genetic engineering of Nang Thom Cho Dao and other indica rice varieties.

Keywords

• Agrobacterium-mediated transformation , Indica rice, Nang Thom Cho Dao (NTCD), Reporter genes