Mopane woodland are a source of valuable resources that contribute substantially to rural economies and nutrition across Southern Africa. However, a number of factors have, of late, brought the sustainability of the mopane woodland resources into question. One of such factors is the difficulty in monitoring of defoliation process within the woodland. In this study we set out to discriminate the levels of change in forest canopy cover detectable after insect defoliation using ground based hyperspectral measurements in mopane woodland. Canopy spectral measurements were taken from three levels of defoliation: Undefoliated (UD), Partly defoliated (PD) and Refoliating plants (R) using ASD FieldSpec HandHeld 2. A pre-filtering approach (ANOVA) was compared with random forest independent variable selector in selecting the significant wavelengths for classification. Furthermore, a backward feature elimination method was used to select optimal wavelengths for discriminating the different levels of defoliation in mopane woodland. Results show that optimal wavelengths located at 707 nm, 710 nm, 711 nm, 712 nm, 713 nm, 714 nm, 727 nm, and 1066 nm were able to discriminate between the three levels of defoliation. The results further show that there was no significant difference in the overall accuracy of classification when random forest variable selector was used 82.42% (Kappa = 0.64) and the pre-filtering approach (ANOVA) 81.21% (Kappa = 0.68) used before building the classification. Overall, the study clearly demonstrated that the dynamic process of defoliation in mopane woodland can be assessed and detected using hyperspectral dataset and effective algorithm for discrimination.