Macleaya cordata is an important medicinal plant to produced benzylisoquinoline alkaloids (BIAs) such as sanguinarine (SAN), Berberine (BBR). This study was aimed at improving the BIAs production and disease resistance of M. cordata. The leaf and stem explants were vacuum infiltration with Agrobacterium for 10min, followed the explants were placed onto co-cultivation medium (100μM acetosyringone) for 3days. Then, the explants were transferred to selected medium (75mg/l kanamycin and 400mg/l timentin). Finally, the putative transformants were verified by GUS histochemical assay and PCR amplification. This is the first report on Agrobacterium-mediated transformation of M. cordata and this method could be providing a regeneration of transgenic M. cordata from leaf and stem segments to increase biomass, alkaloids yield and quality in the future.