Root water uptake is an essential process for terrestrial plants that strongly affects the spatiotemporal distribution of water in vegetated soil. Fast neutron tomography is a recently established non-invasive imaging technique capable to capture the 3D architecture of root systems in situ and even allows for tracking of three-dimensional water flow in soil and roots. We present an in vivo analysis of local water uptake and transport by roots of soil-grown maize plants-for the first time measured in a three-dimensional time-resolved manner. Using deuterated water as tracer in infiltration experiments, we visualized soil imbibition, local root uptake, and tracked the transport of deuterated water throughout the fibrous root system for a day and night situation. This revealed significant differences in water transport between different root types. The primary root was the preferred water transport path in the 13-days-old plants while seminal roots of comparable size and length contributed little to plant water supply. The results underline the unique potential of fast neutron tomography to provide time-resolved 3D in vivo information on the water uptake and transport dynamics of plant root systems, thus contributing to a better understanding of the complex interactions of plant, soil and water.