This study explored the quantitative inversion of the chlorophyll content in Paulownia seedling leaves under drought stress and analyzed the factors influencing the chlorophyll content from multiple perspectives to obtain the optimal model. Paulownia seedlings were selected as the experimental materials for the potted water control experiments. Four drought stress treatments were set up to obtain four types of Paulownia seedlings: one pair of top leaves (T1), two pairs of leaves (T2), three pairs of leaves (T3), and four pairs of leaves (T4). In total, 23 spectral transformations were selected, and the following four methods were adopted to construct the prediction model, select the best spectral preprocessing method, and explore the influence of water bands: partial least squares modeling with all spectral bands (all-band partial least squares, AB-PLS), principal component analysis partial least squares (PCA-PLS), correlation analysis partial least squares (CA-PLS), correlation analysis (water band) partial least squares, ([CA(W)-PLS]), and vegetation index modeling. Based on the prediction accuracy and the uniformity of different leaf positions, the optimal model was systematically explored. The results of the analysis of spectral reflectance showed significant differences at different leaf positions. The sensitive bands of chlorophyll were located near 550 nm, whereas the sensitive bands of water were located near 1440 and 1920 nm. The results of the vegetation index models indicate that the multiple-index models outperformed the single-index models. Accuracy decreased as the number of indicators decreased. We found that different model construction methods matched different optimal spectral preprocessing methods. First derivative spectra (R') was the best preprocessing method for the AB-PLS, PCA-PLS, and CA-PLS models, whereas the inverse log spectra (log(1/R)) was the best preprocessing method for the CA(W)-PLS model. Among the 14 indices, the green normalized difference vegetation index (GNDVI) was most correlated with the chlorophyll content sensitivity indices, and the water index (WI) was most correlated with the water sensitive indices. At the same time, the water band affected the cross validation accuracy. When characteristic bands were used for modeling, the cross validation accuracy was significantly increased. In contrast, when vegetation indices were used for modeling, the accuracy of the cross validation increased slightly but its predictive ability was reduced; thus, these changes could be ignored. We found that leaf position also affected the prediction accuracy, with the first pair of top leaves exhibiting the worst predictive ability. This was a bottleneck that limited predictive capability. Finally, we found that the CA(W)-PLS model was optimal. The model was based on 23 spectral transformations, four PLS construction methods, water bands, and different leaf positions to ensure systematicity, stability, and applicability.