Anthocyanin can improve the stress tolerance and disease resistance of winter wheat to a certain extent, so timely and accurate monitoring of anthocyanin content is crucial for the growth and development of winter wheat. This study measured the ground-based hyperspectral reflectance and the corresponding anthocyanin concentration at four key growth stages—booting, heading, flowering, and filling—to explore the spectral detection of anthocyanin in winter wheat leaves. Firstly, the first-order differential spectra (FDS) are obtained by processing based on the original spectra (OS). Then, sensitive bands (SBS), the five vegetation indices for optimal two-band combinations (VIo2), and the five vegetation indices for optimal three-band combinations (VIo3) were selected from OS and FDS by band screening methods. Finally, modeling methods such as RF, BP, and KELM, as well as models optimized by genetic algorithm (GA), were used to estimate anthocyanin content at different growth stages. The results showed that (1) among all the models, the GA_RF had incredible performance, VIo3 was the superior parameter for estimating anthocyanin values, and the model GA_RF of FDS data based on VIo3 for the filling stage (Rv2 = 0.950, RMSEv = 0.005, RPDv = 4.575) provided the best estimation of anthocyanin. (2) the first-order differential processing could highlight the degree of response of SBS, VIo2, and VIo3 to the anthocyanin values. The model performances of the FDS were better than that of OS on the whole, and the Rv2 of the optimal models of FDS were all greater than 0.89. (3) GA had optimizing effects on the RF, BP, and KELM, and overall, the GA models improved the R2 by 0.00%-18.93% compared to the original models. These results will provide scientific support for the use of hyperspectral techniques to monitor anthocyanin in the future.