Abstract: Vacuoles are ubiquitous organelles in the fungal and plant kingdoms. They serve a variety of functions and are important for cell homeostasis. A constant turnover of proteins and membranes makes vacuoles dynamic organelles. Various transport pathways share the vacuole as their joint destination. The trafficking pathways are regulated independently. In yeast cells many components of the protein and membrane transport machinery are known. Recent years have seen much progress in our understanding of the protein-sorting pathways and the biogenesis of this organelle. Improvements of our understanding of the vesicular transport pathways and vacuolar membrane fusion are reviewed.