Global food security is critical to eliminating hunger and malnutrition. In the changing climate, farmers in developing countries must adopt technologies and farming practices such as precision agriculture (PA). PA-based approaches enable farmers to cope with frequent and intensified droughts and heatwaves, optimising yields, increasing efficiencies, and reducing operational costs. Biophysical parameters such as Leaf Area Index (LAI), Leaf Chlorophyll Content (LCab), and Canopy Chlorophyll Content (CCC) are essential for characterising field-level spatial variability and thus are necessary for enabling variable rate application technologies, precision irrigation, and crop monitoring. Moreover, robust machine learning algorithms offer prospects for improving the estimation of biophysical parameters due to their capability to deal with non-linear data, small samples, and noisy variables. This study compared the predictive performance of sparse Partial Least Squares (sPLS), Random Forest (RF), and Gradient Boosting Machines (GBM) for estimating LAI, LCab, and CCC with Sentinel-2 imagery in Bothaville, South Africa and identified, using variable importance measures, the most influential bands for estimating crop biophysical parameters. The results showed that RF was superior in estimating all three biophysical parameters, followed by GBM which was better in estimating LAI and CCC, but not LCab, where sPLS was relatively better. Since all biophysical parameters could be achieved with RF, it can be considered a good contender for operationalisation. Overall, the findings in this study are significant for future biophysical product development using RF to reduce reliance on many algorithms for specific parameters, thus facilitating the rapid extraction of actionable information to support PA and crop monitoring activities.