| Summary: | The estimation of anthocyanin (Anth) content is very important for observing the physiological state of plants under environmental stress. The objective of this study was to estimate the Anth of maize leaves at different growth stages based on remote sensing methods. In this study, the hyperspectral reflectance and the corresponding Anth of maize leaves were measured at the critical growth stages of nodulation, tasseling, lactation, and finishing of maize. First-order differential spectra (FD) were derived from the original spectra (OS). First, the spectral parameters highly correlated with Anth were selected. A total of two sensitive bands (R<sub>λ</sub>), five classical vegetation indices (VI<sub>S</sub>), and six optimized vegetation indices (VI<sub>C</sub>) were selected from the original and first-order spectra. Then, univariate regression models for Anth estimation (Anth-UR models) and multivariate regression models for estimating anthocyanins (Anth-MR models) were constructed based on these parameters at different growth stages of maize. It was shown that the first-order spectral conversion could effectively improve the correlation between R<sub>λ</sub>, VI<sub>C</sub>, and Anth, and VI<sub>C</sub> are usually more sensitive to Anth than VI<sub>S</sub>. In addition, the overall performance of Anth-MR models was better than that of Anth-UR models. Among them, Anth-MR models with the combination of three types of spectral parameters (FD(R<sub>λ</sub>) + OS_VI<sub>C</sub> + FD_VI<sub>C</sub>/VI<sub>S</sub>) as inputs had the best overall performance. Moreover, different growth stages had an impact on the Anth estimation models, with tasseling and lactation stages showing better results. The best-performing Anth-MR models for these two growth stages were as follows. For the tasseling stage, the best model was the FD(R<sub>λ</sub>) + OS_VI<sub>C</sub> + VI<sub>S</sub>-based SVM model, with an R<sup>2</sup> of 0.868, RMSE of 0.007, and RPD of 2.19. For the lactation stage, the best-performing model was the FD(R<sub>λ</sub>) + OS_VI<sub>C</sub> + FD_VI<sub>C</sub>-based RF model, with an R<sup>2</sup> of 0.797, RMSE of 0.007, and RPD of 2.24. These results will provide a scientific basis for better monitoring of Anth using remote sensing hyperspectral techniques.
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