Nitrogen (N) remobilization is a critical process that provides substantial N to winter wheat grains for improving yield productivity. However, our understanding on the N remobilization efficiency (NRE) is still limited due to the lack of efficient, repeatable approaches for evaluating the NRE. In this study, we provided a proof of concept for estimating N concentration and assessing N remobilization using hyperspectral data of individual organs, which offers a non-chemical and low-cost approach to screen germplasms for an optimal NRE in drought-resistance breeding.