The in vivo features of the absorption of leaf photosynthetic and photo-protective pigments are closely linked to the leaf spectrum in the 400–800 nm regions. However, this information is difficult to obtain because the overlapping leaf pigments can mask the contribution of individual pigments to the leaf spectrum. Here, to limit the masking phenomenon between these pigments, the separation technology for leaf spectral overlapping was employed in the PROSPECT model with the ZJU dataset. The main results of this study include the following aspects: (1) the absorption coefficients of separated chlorophyll a and b, carotenoids and anthocyanins in the leaf in vivo display the physical principles of forming an absorption spectrum similar to those in an organic solution; (2) the differences in the position of each absorption peak of pigments between the leaf in vivo and in an organic solution can be described by a spectral displacement parameter; and (3) the overlapping characteristics between the separated pigments in the leaf in vivo are clearly drawn by a range of absorption feature (RAF) parameter. Moreover, the absorption coefficients of the separated pigments were successfully applied in leaf spectral modeling and pigment retrieval. The results show that the separated multiple pigment absorption coefficients from the leaf spectrum in vivo are effective and provide a framework for future refinements in describing leaf optical properties.

The PROSPECT leaf optical radiative transfer models, including PROSPECT-MP, have addressed the contributions of multiple photosynthetic pigments (chlorophyll a and b, and carotenoids) to leaf optical properties, but photo-protective pigment (anthocyanins), another important indicator of vegetation physiological and ecological functions, has not been simultaneously combined within a leaf optical model. Here, we present a new calibration and validation of PROSPECT-MP+ that separates the contributions of multiple photosynthetic and photo-protective pigments to leaf spectrum in the 400–800 nm range using a new empirical dataset that contains multiple photosynthetic and photo-protective pigments (LOPEX_ZJU dataset). We first provide multiple distinct in vivo individual photosynthetic and photo-protective pigment absorption coefficients and leaf average refractive index of the leaf interior using the LOPEX_ZJU dataset. Then, we evaluate the capabilities of PROSPECT-MP+ for forward modelling of leaf directional hemispherical reflectance and transmittance spectra and for retrieval of pigment concentrations by model inversion. The main result of this study is that the absorption coefficients of chlorophyll a and b, carotenoids, and anthocyanins display the physical principles of absorption spectra. Moreover, the validation result of this study demonstrates the potential of PROSPECT-MP+ for improving capabilities in remote sensing of leaf photosynthetic pigments (chlorophyll a and b, and carotenoids) and photo-protective pigment (anthocyanins).