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Nitrogen content is a crucial index for crop growth diagnosis and the exact estimation of nitrogen content is of great significance for grasping crop growth status in real-time. This paper takes winter wheat as the study object and the precision agriculture demonstration area of the Jiaozuo Academy of Agricultural and Forestry Sciences in Henan Province as the research area. The hyperspectral reflectance data of the wheat canopy in different growth periods are obtained with the ASD ground object hyperspectral instrument, and the original canopy spectral data are preprocessed by fractional differential and continuous wavelet transform; then, the vegetation indices are established, correlation analysis with nitrogen content is conducted, and the fractional differential spectra are selected; finally, based on the wavelet energy coefficient and the vegetation indices with strong correlations, the methods of support vector machine (SVM), ridge regression, stepwise regression, Gaussian process regression (GPR), and the BP neural network are used to construct the estimation model for nitrogen content in wheat at different growth stages. By adopting the  R 2  and root mean square error (RMSE) indices, the best nitrogen content estimation model at every growth stage is selected. The overall analysis of the nitrogen content estimation effect indicated that for the four growth periods, the maximum modeling and validation  R 2  of the nitrogen content estimation models of the SVM, ridge regression, stepwise regression, GPR, and BP neural network models reached 0.95 and 0.93, the average reached 0.76 and 0.71, and the overall estimation effect was good. The average values of the modeling and validation  R 2  of the nitrogen content estimation model at the flag picking stage were 0.85 and 0.81, respectively, which were 37.10% and 44.64%, 1.19% and 3.85%, and 14.86% and 17.39% higher than those at the jointing stage, flowering stage, and filling stage, respectively. Therefore, the model of the flag picking stage has higher estimation accuracy and a better estimation effect on the nitrogen content. For the different growth stages, the optimal estimation models of nitrogen content were different. Among them, continuous wavelet transform combined with the BP neural network model can be the most effective method for estimating the N content in wheat at the flagging stage. The paper provides an effective method for estimating the nitrogen content in wheat and a new idea for crop growth monitoring. [ABSTRACT FROM AUTHOR]