Selective inhibition of carotenoid cleavage dioxygenases : phenotypic effects on shoot branching

Members of the carotenoid cleavage dioxygenase family catalyse the oxidative cleavage of carotenoids at various chain positions, leading to the formation of a wide range of apocarotenoid signalling molecules. To explore the functions of this diverse enzyme family, we have used a chemical genetic approach to design selective inhibitors for different classes of carotenoid cleavage dioxygenase. A set of 18 arylalkyl-hydroxamic acids was synthesised in which the distance between an iron-chelating hydroxamic acid and an aromatic ring was varied; these compounds were screened as inhibitors of four different enzyme classes, either in vitro or in vivo. Potent inhibitors were found that selectively inhibited enzymes that cleave carotenoids at the 9,10 position; 50% inhibition was achieved at sub-micromolar concentrations. Application of certain inhibitors at 100 microM to Arabidopsis node explants or whole plants led to increased shoot branching, consistent with inhibition of 9,10-cleavage