Freezing injury may cause irreversible damage to wheat (Triticum aestivum L) tissues and can significantly reduce yield and quality. Therefore, quick and non-destructively estimating the degree of frost damage for formulating anti-freezing protection strategies and preventing frost damage is very crucial. In this study, we obtained hyperspectral images of wheat leaves for accurate identification of frost damage. A remote-controlled Unmanned Ground Vehicle (UGV) equipped with an imaging spectral camera was used to capture the hyperspectral images of frost-damaged wheat leaves. We compared the efficiency of two methods (the one without removal of weeds, and the other is to remove the corresponding area of weeds from the hyperspectral image by Deeplab V3+) for estimation of wheat freezing damage degree by using four different algorithms; Support Vector Machine Classification (SVM), Mahalanobis Distance Classification (MaD), Minimum Distance Classification (MiD), and Maximum Likelihood Classification (ML). We found that, Deeplab V3+ can efficiently identify the weeds from hyperspectral images, as the overall accuracy (OA) values of different algorithms were high in images with weeds removal as compared to the values in weeds containing images. Further, applying ML model after weeds removal have high OA (93.26 %) as compared to the other models. Thus, using Deeplab V3+ and ML can be a potential approach to identify the freezing injury in wheat for sustainable agricultural productivity.