Timely diagnosis of sugar beet above-ground biomass (AGB) is critical for the prediction of yield and optimal precision crop management. This study established an optimal quantitative prediction model of AGB of sugar beet by using hyperspectral data. Three experiment campaigns in 2014, 2015 and 2018 were conducted to collect ground-based hyperspectral data at three different growth stages, across different sites, for different cultivars and nitrogen (N) application rates. A competitive adaptive reweighted sampling (CARS) algorithm was applied to select the most sensitive wavelengths to AGB. This was followed by developing a novel modified differential evolution grey wolf optimization algorithm (MDE–GWO) by introducing differential evolution algorithm (DE) and dynamic non-linear convergence factor to grey wolf optimization algorithm (GWO) to optimize the parameters c and γ of a support vector machine (SVM) model for the prediction of AGB. The prediction performance of SVM models under the three GWO, DE–GWO and MDE–GWO optimization methods for CARS selected wavelengths and whole spectral data was examined. Results showed that CARS resulted in a huge wavelength reduction of 97.4% for the rapid growth stage of leaf cluster, 97.2% for the sugar growth stage and 97.4% for the sugar accumulation stage. Models resulted after CARS wavelength selection were found to be more accurate than models developed using the entire spectral data. The best prediction accuracy was achieved after the MDE–GWO optimization of SVM model parameters for the prediction of AGB in sugar beet, independent of growing stage, years, sites and cultivars. The best coefficient of determination (R2), root mean square error (RMSE) and residual prediction deviation (RPD) ranged, respectively, from 0.74 to 0.80, 46.17 to 65.68 g/m2 and 1.42 to 1.97 for the rapid growth stage of leaf cluster, 0.78 to 0.80, 30.16 to 37.03 g/m2 and 1.69 to 2.03 for the sugar growth stage, and 0.69 to 0.74, 40.17 to 104.08 g/m2 and 1.61 to 1.95 for the sugar accumulation stage. It can be concluded that the methodology proposed can be implemented for the prediction of AGB of sugar beet using proximal hyperspectral sensors under a wide range of environmental conditions.