Significance Vacuoles play crucial roles in plant growth and adaptation to the environment. However, the mechanisms responsible for transporting membranes and contents to plant vacuoles remain largely uncharacterized, and the pathways and compartments involved are not fully charted. We report on the characterization of 17 vacuolar trafficking mutants that define a set of 13 cellular factors involved in transport of soluble vacuolar proteins in Arabidopsis thaliana . We show that two of these factors, the GARP tethering complex subunits VPS51 and VPS54, reside in an ER- and microtubule-associated compartment (EMAC) and that VPS51 interacts with the motor domain of kinesins, revealing an unknown compartment of the vacuolar pathway and suggesting that microtubules and kinesins participate in vacuolar trafficking in plants.