Spectral reflectance of maple, chestnut and beech leaves in a wide range of pigment content and composition was investigated to devise a nondestructive technique for total carotenoid (Car) content estimation in higher plant leaves. Reciprocal reflectance in the range 510 to 550 nm was found to be closely related to the total pigment content in leaves. The sensitivity of reciprocal reflectance to Car content was maximal in a spectral range around 510 nm; however, chlorophylls (Chl) also affect reflectance in this spectral range. To remove the Chl effect on the reciprocal reflectance at 510 nm, a reciprocal reflectance at either 550 or 700 nm was used, which was linearly proportional to the Chl content. Indices for nondestructive estimation of Car content in leaves were devised and validated. Reflectances in three spectral bands, 510 ± 5 nm, either 550 ± 15 nm or 700 ± 7.5 nm and the near infrared range above 750 nm are sufficient to estimate total Car content in plant leaves nondestructively with a root mean square error of less than 1.75 nmol/cm2.