Transformation of Arabidopsis with
pCCA1::LUC+
Agrobacterium tumefaciens (Gv3101) transformed with appropriate
constructs (Ti plasmid: pPCVH containing LUC+ ; helper plasmid:
pM90RK; CCA1:luc binary vector), a gift from A. Millar
(University of Edinburgh) were inoculated into 5 ml Luria Bertani
cultures containing gentamycin (25 µg ml-1) and
ampicillin (50 µg ml-1) two days before
transformation. After cultures were left overnight at 30 °C, 2.5 ml was
sub-cultured into a 50 ml culture, and then grown at 30 °C overnight.
The culture was generously painted onto whole plants (leaves, secondary
stems and inflorescences) with an autoclaved paintbrush. Plants were
covered in a cling film tent for two days to increase humidity.