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| Description: (a) Cytokinin Opine production synthesis Auxin production T-DNA left border 1 Tumor-inducing \ (Ti) plasmid Virulence Genes for opine catabolisrn region arr' T—DNA right border conjugative ransfer % r Reengineenng of Ti plasmid separates sequences Mia transier of T-DNA From the T—DNA itselft Genes for conjugative transfer Virulence t Kan" reg IOI'I (bacterial selectable marker) Tl plasmid (T-region removed) cri ( "Drsarmed" plasmid contains genes required for ‘ virulence and conjugatrve transfer; lacking T—regron, i it is no longer able to Induce crown gall disease. (b) Disarmed plasmid and vector are transferred into an Agrobacterium. Agmbacteriurn Plant cell Disarmed Ti plasmid Transformation vector Plant selectable marker (e.g.. herbicide resistance) Constitutive promoter T-DNA left border f 3‘ transcriptional terminator Mcs (inserted gene of interest) T-DNA right border T-region Transformation vector Amp’l (bacterial selectable marker) ori Transformation vector contains T-region flanked by right and left border sequences. (t) fik Grow plantlets Transgenic plant Culture cells containing herbicide and, alter selection, regenerated into ‘r Infected plant cells are grown on selectable media l transgenic plants, l Genes on disarrréd plasmid produce conjugative and virulence proteins that act in trans on T-DNA border sequences of transfomtation vector to effect transfer of Y-DNA, which contains the inserted gene of interest into plant cell.
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