With current and upcoming imaging spectrometers, automated band analysis techniques are needed to enable efficient identification of most informative bands to facilitate optimized processing of spectral data into estimates of biophysical variables. This paper introduces an automated spectral band analysis tool (BAT) based on Gaussian processes regression (GPR) for the spectral analysis of vegetation properties. The GPR-BAT procedure sequentially backwards removes the least contributing band in the regression model for a given variable until only one band is kept. GPR-BAT is implemented within the framework of the free ARTMO's MLRA (machine learning regression algorithms) toolbox, which is dedicated to the transforming of optical remote sensing images into biophysical products. GPR-BAT allows (1) to identify the most informative bands in relating spectral data to a biophysical variable, and (2) to find the least number of bands that preserve optimized accurate predictions. To illustrate its utility, two hyperspectral datasets were analyzed for most informative bands: (1) a field hyperspectral dataset (400–1100 nm at 2 nm resolution: 301 bands) with leaf chlorophyll content (LCC) and green leaf area index (gLAI) collected for maize and soybean (Nebraska, US); and (2) an airborne HyMap dataset (430–2490 nm: 125 bands) with LAI and canopy water content (CWC) collected for a variety of crops (Barrax, Spain). For each of these biophysical variables, optimized retrieval accuracies can be achieved with just 4 to 9 well-identified bands, and performance was largely improved over using all bands. A PROSAIL global sensitivity analysis was run to interpret the validity of these bands. Cross-validated View the MathML source (NRMSECV) accuracies for optimized GPR models were 0.79 (12.9%) for LCC, 0.94 (7.2%) for gLAI, 0.95 (6.5%) for LAI and 0.95 (7.2%) for CWC. This study concludes that a wise band selection of hyperspectral data is strictly required for optimal vegetation properties mapping.