High throughput In Vitro Bioassays

The ubiquitin E3 ligases are known to have pleiotropic functions through targeting several substrates and thereby affecting several regulatory pathways. We therefore set out to screen their gene expression patterns in hormonal perturbations. Mutants for ABA responding E3 ligases were used to confirm their hormonal associations using an automated bioassay in an image based phenotyping system. In this system hormone concentration gradients were applied in multiwell plates that were imaged by RGB and FluorCam. Thereby we could identified ubiquitin E3 ligases that were either sensitive or resistant to ABA during the early stages of development.