Experiences can alter functional properties of neurons in primary sensory neocortex but it is poorly understood how stimulus-reward associations contribute to these changes. Using in vivo two-photon calcium imaging in mouse primary visual cortex (V1), we show that association of a directional visual stimulus with reward results in broadened orientation tuning and sharpened direction tuning in a stimulus-selective subpopulation of V1 neurons. Neurons with preferred orientations similar, but not identical to, the CS+ selectively increased their tuning curve bandwidth and thereby exhibited an increased response amplitude at the CS+ orientation. The increase in response amplitude was observed for a small range of orientations around the CS+ orientation. A nonuniform spatial distribution of reward effects across the cortical surface was observed, as the spatial distance between pairs of CS+ tuned neurons was reduced compared with pairs of CS- tuned neurons and pairs of control directions or orientations. These data show that, in primary visual cortex, formation of a stimulus-reward association results in selective alterations in stimulus-specific assemblies rather than population-wide effects.