Development of Agrobacterium-mediated in planta transformation method through coleoptile in rice

Abstract Genetic modification of rice is mainly carried out by Agrobacterium-mediated transformation of callus accompanied by tissue culture. It is time consuming, laborious and useless for cultivars that are unable to induce callus. In this paper, we report a novel gene transfer to pull out primary leaf followed by infection of shoot apical meristem in the coleoptile with Agrobacterium tumefaciens EHA105 culture harboring pCAMBIA1301-RD29A-AtDREB1A. Primary leaves were pulled out from rice seedlings grown in Petri dishes and soil conditions, and Agrobacterium culture were injected into the coleoptile with a micro syringe. Treated plants were co-cultured in dark condition for 48 hours and washed. These plants were regenerated into transgenic plants, and the transgene was transferred to the next generation, the transgenic plants exhibited tolerance to cold stress. This gene transfer through rice coleoptile is a simple, efficient and cultivar-free method that does not require tissue culture.