The trans-Golgi network (TGN) is a tubular-vesicular organelle that matures from the trans cisternae of the Golgi apparatus. In plants, the TGN functions as a central hub for three trafficking pathways: the secretory pathway, the vacuolar trafficking pathway and the endocytic pathway. Here, we describe a novel TGN-localized membrane protein, CONTINUOUS VASCULAR RING (COV1), that is crucial for TGN function in Arabidopsis. The COV1 gene was originally identified from the stem vascular patterning mutant of Arabidopsis thaliana. However, the molecular function of COV1 was not identified. Fluorescently tagged COV1 proteins co-localized with the TGN marker proteins, SYNTAXIN OF PLANTS 4 (SYP4) and vacuolar-type H(+)-ATPase subunit a1 (VHA-a1). Consistently, COV1-localized compartments were sensitive to concanamycin A, a specific inhibitor of VHA. Intriguingly, cov1 mutants exhibited abnormal Golgi morphologies, including a reduction in the number of Golgi cisternae and a reduced association between the TGN and the Golgi apparatus. A deficiency in COV1 also resulted in a defect in vacuolar protein sorting, which was characterized by the abnormal accumulation of storage protein precursors in seeds. Moreover, we found that the development of an idioblast, the myrosin cell, was abnormally increased in cov1 leaves. Our results demonstrate that the novel TGN-localized protein COV1 is required for Golgi morphology, vacuolar trafficking and myrosin cell development.