Relationships between leaf pigment content and spectral reflectance across a wide range of species, leaf structures and developmental stages

Leaf pigment content can provide valuable insight into the physiological performance of leaves. Measurement of spectral reflectance provides a fast, nondestructive method for pigment estimation. A large number of spectral indices have been developed for estimation of leaf pigment content. However, in most cases these indices have been tested for only one or at most a few related species and thus it is not clear whether they can be applied across species with varying leaf structural characteristics. Our objective in this study was to develop spectral indices for prediction of leaf pigment content that are relatively insensitive to species and leaf structure variation and thus could be applied in larger scale remote-sensing studies without extensive calibration. We also quantified the degree of spectral interference between pigments when multiple pigments occur within the same leaf tissue. We found that previously published spectral indices provided relatively poor correlations with leaf chlorophyll content when applied across a wide range of species and plant functional types. Leaf surface reflectance appeared to be the most important factor in this variation. By developing a new spectral index that reduces the effect of differences in leaf surface reflectance, we were able to significantly improve the correlations with chlorophyll content. We also found that an index based on the first derivative of reflectance in the red edge region was insensitive to leaf structural variation. The presence of other pigments did not significantly affect estimation of chlorophyll from spectral reflectance. Previously published carotenoid and anthocyanin indices performed poorly across the whole data set. However, we found that the photochemical reflectance index (PRI, originally developed for estimation of xanthophyll cycle pigment changes) was related to carotenoid/chlorophyll ratios in green leaves. This result has important implications for the interpretation of PRI measured at both large and small scales. Our results demonstrate that spectral indices can be applied across species with widely varying leaf structure without the necessity for extensive calibration for each species. This opens up new possibilities for assessment of vegetation health in heterogeneous natural environments.