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Biophysical parameters such as leaf chlorophyll content (LCC) and leaf area index (LAI) are standard vegetation products that can be retrieved from Earth observation imagery. This paper introduces a new machine learning regression algorithms (MLRAs) toolbox into the scientific Automated Radiative Transfer Models Operator (ARTMO) software package. ARTMO facilitates retrieval of biophysical parameters from remote observations in a MATLAB graphical user interface (GUI) environment. The MLRA toolbox enables analyzing the predictive power of various MLRAs in a semiautomatic and systematic manner, and applying a selected MLRA to multispectral or hyperspectral imagery for mapping applications. It contains both linear and nonlinear state-of-the-art regression algorithms, in particular linear feature extraction via principal component regression (PCR), partial least squares regression (PLSR), decision trees (DTs), neural networks (NNs), kernel ridge regression (KRR), and Gaussian processes regression (GPR). The performance of multiple implemented regression strategies has been evaluated against the SPARC dataset (Barrax, Spain) and simulated Sentinel-2 (8 bands), CHRIS (62 bands) and HyMap (125 bands) observations. In general, nonlinear regression algorithms (NN, KRR, and GPR) outperformed linear techniques (PCR and PLSR) in terms of accuracy, bias, and robustness. Most robust results along gradients of training/validation partitioning and noise variance were obtained by KRR while GPR delivered most accurate estimations. We applied a GPR model to a hyperspectral HyMap flightline to map LCC and LAI. We exploited the associated uncertainty intervals to gain insight in the per-pixel performance of the model.