Polyamines (PAs) are unique polycationic metabolites, which modulate plants' growth, development, and stress responses. As polycations, PAs interfere with cationic transport systems as ion channels and ionotropic pumps. Here, we describe the application of two techniques, MIFE to study the effects of PAs on cation fluxes in vivo and conventional patch-clamp to evaluate the PA blockage of ion currents in isolated plant vacuoles. Preparation of vacuoles for patch-clamp assays is described and solutions and voltage protocols are given, which allow separate recordings of major vacuolar channel currents and quantify their blockage by PAs.
Keywords: Calcium; H+ pump; Ion channel; MIFE; Patch-clamp; Plasma membrane; Root; Vacuole.