BACKGROUND Strigolactones (SLs) are plant hormones that play various roles in plant development. The chemical stability of SLs depends on the solvent, pH, and the presence of nucleophiles. Hydrolysis leads to detachment of the butenolide ring, and plays a crucial role in the initial stages of the signal-transduction process occurring between the receptor and the SL signaling molecule. RESULTS To date, two different mechanisms have been proposed for SL hydrolysis. Results obtained from kinetic, thermodynamic, and mass spectral data of the reaction between widely used synthetic strigolactone analog GR24 and seven different nucleophiles, demonstrate that the reaction proceeds via the Michael addition-elimination mechanism. CONCLUSIONS This paper provides valuable information on the chemical stability of GR24 in different plant growing media and buffers. Such information is valuable for scientists using GR24 treatments to study SL-regulated processes in plants.