An efficient Agrobacterium -mediated transformation and regeneration protocol for aromatic black rice, Oryza sativa L. (cv. Chakhao Poireiton and Chakhao Amubi)

Crop-improvement efforts for Chakhao black rice ( Oryza sativa L.) through genetic engineering have not made much progress due to the absence of a protocol for callus induction and regeneration. We developed a tissue culture protocol for the Agrobacterium -mediated transformation of Chakhao Amubi and Chakhao Poireiton cultivars of rice using mature seeds as the explants. The transformation protocol is simple and can be carried out in laboratories with basic equipment for tissue culture. The effect of temperature, growth regulators, gelling agents, and the duration of infection on different stages of the protocol is also described. Transgenic rice seedlings can be obtained in 69–80 days with > 30% transformation efficiency using the optimized method for each genotype.