Agrobacterium Leaf Disk Transformation

1. Agrobacterium Leaf Disk Transformation Microprojectile Callus Transformation Cocultivate leaｆ disk with Agrobacterium Initiate & proliferate embryogenic callus 1 – 2 days 8 – 12 weeks Wash leaf disk Microprojectile bombardment Callus growth without selection Antibiotic treatment to kill Agrobacterium & select transformed plant cells 2 – 4 weeks 4 days Callus growth under escape-free selection Surviving leaf disks onto fresh selection/ regeneration medium 8 – 12 weeks 6 – 10 weeks Plant regeneration and rooting on escape -free selection Root excised plantlets on selection 8 – 12 weeks 4 – 6 weeks Potted transgenic plants Potted transgenic plants ca 12 – 20 weeks Total time in culture Total time in culture ca 24 – 36 weeks Fig. 1.1. Typical transformation regimens of Agrobacterium-mediated and microparticle bombardment methods 5).