This paper assesses the capability of hyperspectral remote sensing to estimate chlorophyll a (Chl a), chlorophyll b (Chl b), and chlorophyll a + b (Chl a + b) concentration at cherry leaf scale using a variety of spectral variables during the growth season. A field experiment was conducted in cherry orchard. The leaf reflectance spectra of cherry plants were acquired within 350-1050 nm wavelengths. A variety of spectral variables were mathematically computed based on the leaf spectra and transformation of reflectance spectra. The relationships between all of spectral variables and chlorophyll concentration were discussed. Estimating Chl a, Chl b, and Chl a + b concentration by stepwise linear regression method and curve estimation method were carried out. Results demonstrated that the best spectral variable for prediction of chlorophyll concentration was the new spectral variable (the first derivative of log (1/R 741 ) and D 751 /D 511 ), with the root mean square error prediction (RMSEP) of 4.802 mg L −1 for Chl a concentration, 1.659 mg L −1 for Chl b concentration, and 6.419 mg L −1 for Chl a + b concentration. It should be noted that spectral variables such as D 715 /D 705 , EBFR, D 705 /D 722 , and BND showed a good performance with the RMSEP of 5. 768, 7.838, 12.146, and 14.437 mg L −1 for Chl a concentration, 1.795, 1.985, 1.765, and 3.164 mg L −1 for Chl b concentration, and 7. 935, 11.49, 17.99, and 21.79 mg L −1 for Chl a + b concentration respectively. Further investigation is needed to evaluate the effectiveness of such techniques on other orchard varieties or at the canopy level.