The compartment specific distribution of ascorbate in plants is of great importance for plant development, growth and defense as this multifunctional metabolite plays important roles in the detoxification of reactive oxygen species (ROS), redox signaling, modulation of gene expression and is important for the regulation of enzymatic activities. Even though changes in ascorbate contents during plant growth and various stress conditions are well documented and the roles of ascorbate in plant defense during abiotic stress conditions are well established, still too little is known about its compartment specific roles during plant development and defense. This mini-review focuses on the subcellular distribution of ascorbate in plants and describes different methods that are currently used to study its compartment specific distribution. Finally, it will also briefly discuss data available on compartment specific changes of ascorbate during some abiotic stress conditions such as high light conditions and exposure to ozone.