The purpose of this study was to transform rolC::Hd3a-GFP by Agrobacterium tumefaciens on black rice (Oryza sativa L. cv. Cempo Ireng). The callus formation were induced from scutellum using two different media, 2N 6 and 2,4-dichlorophenoxyacetic acid (2,4-D). PCR analysis using specific primers for Hd3a and hpt was performed to determine the stability of rolC::Hd3a-GFP construct before co-cultivation of callus and Agrobacterium tumefaciens. This study indicated that black rice callus induction on 2N 6 medium responded faster than that on MS 2,4-D medium and generated friable calli for Agrobaterium-mediated transformation. The screening result showed that several calli were competent to regenerate on medium containing hygromysin and demonstrated Hd3a insertion gene.