New 3-D Fluorescence Spectral Indices for Multiple Pigment Inversions of Plant Leaves via 3-D Fluorescence Spectra

Abstract

High-sensitivity fluorescence monitoring has been widely used in agriculture and environmental science. However, the active fluorescence detection information of leaf segments mainly focuses on total chlorophyll, and the fluorescence information of chlorophyll a, chlorophyll b, and some other pigments has not been explored. This only considers the fluorescence spectrum characteristics at a single wavelength or the fluorescence integral from a range of wavelength regions and does not completely consider the linkage relation between the excitation, emission, and interference information. In this paper, the three-dimensional fluorescence spectrum, containing the excitation and emission fluorescence spectra, and the corresponding multiple pigment characteristics from the upgraded LOPEX_ZJU database were collected. The linkages of excitation and emission of the three-dimensional fluorescence spectra of these pigments were analyzed for the newly built multiple pigment 3-D fluorescence spectral indices (3-D FSIs), including those of chlorophyll a, chlorophyll b, carotenoid, anthocyanin, and flavonoid 3-D FSIs. Then, these pigment inversion models were established and validated. The results show that the 3-D FSIs performances for the photosynthetic pigment content inversion (including chlorophyll a and b, and carotenoids) were much better than those for the photo-protective pigments (including anthocyanins and flavonoids) from the 3-D fluorescence spectra of these plant leaves. Here, the 3-D fluorescence normalization index (FNI ((F430,690 − F430,763)/(F430,690 + F430,763))) for the chlorophyll a inversion model has a high accuracy, the RMSE is 2.96 μg/cm2, and the 3-D fluorescence reciprocal difference index (FRI (F650,704/F650,668) for the chlorophyll b model has an encouraging RMSE (2.01 μg/cm2). The RMSE of the 3-D fluorescence ratio index (FRI (F500,748/F500,717)) for the carotenoid inversion is 3.77 μg/cm2 RMSE. Only FRI (F370,615/F370,438) was selected for the modeling and validating evaluation of the leaf Flas content inversion, but the evaluation metrics were not good, with an RMSE (151.13 μg/cm2). For Ants, although there was a 3-D FSI (FRDI (1/F540,679 − 1/F540,557)), and its evaluation metrics, with an RMSE (2.8 μg/cm2), were at or over 0.05 level, the validating evaluation metric VC (98.3577%) was not encouraging. These results showed that fluorescence, as a nondestructive and efficient detection method, could determine the contents of chlorophyll a, chlorophyll b, and carotenoid in plant leaves, providing a new method to detect plant information. It can also provide a potential chance for the fluorescence images of fine photo-protective pigments, especially chlorophyll a and b, using the special active fluorescence excitation light source and a few fluorescence imaging channels from the optimal FSIs.