Citrus is one of the most economically valuable fruit trees in the world, for which leaf chlorophyll content (LCC) serves as a crucial indicator for evaluating its growth and health status. However, the quantitative estimation of LCC using remote-sensing techniques is still challenging owing to unclear sensitive spectral ranges, baseline drift and overlapping spectrum peaks. To resolve these issues, we clarified the spectral response characteristics of citrus LCC using fractional-order derivatives (FOD) and continuous wavelet transform (CWT) methods to determine its sensitive spectral range with in situ full-spectrum leaf hyperspectral data. We proposed a novel method for estimating the LCC of citrus by combining an ensemble learning regression model based on Hyperopt optimization (H-ELR) with partial least squares regression (PLSR) using the ultra-high dimensional feature variables produced by dual- and tri-band combination strategies. We evaluated the retrieval performance of LCC between the FOD- and CWT-based optimal spectral feature variables. Besides, we further examined the feasibility of improving the estimation accuracy of LCC by the combination of their optimal feature variables. Finally, we evaluated the effect of the spectral curve trend changes and different dimensional spectral features on LCC estimation. The results showed that: (1) The FOD- and CWT-based methods improved the correlation between original spectral reflectance and LCC, with the correlation coefficient increasing by 0.046 and 0.054, respectively. We confirmed that 425–740 nm is the optimal spectral range for LCC estimation. (2) We found that the 0.9 order derivative Tri-band index (TBI2 (R571, R1697, R740)) constructed by combining the sensitive spectral bands of leaf water content achieved a high-precision LCC estimation (R2 = 0.876). In addition, the MERIS terrestrial chlorophyll index (MTCI) constructed based on the scale6 of CWT also gained good retrieval accuracy (R2 = 0.806). (3) We demonstrated that a combination of FOD-based and CWT-based sensitive spectral features improves estimation accuracies (R2 = 0.891) of LCC and revealed that the reflectance peaks and slope peaks at 550 and 750 nm are essential variables for predicting the citrus LCC. (4) The combination of PLSR and H-ELR model provided a good retrieval performance of citrus LCC with the kurtosis (γ = 3.2) and skewness (Sk = 0.066) of the residual prediction values. Our proposed method can provide a scientific basis for estimating LCC and other physiological parameters of citrus and other crop types, which is also important to optimize agricultural management practices and improve crop yield.