The objective of this study was to rapidly evaluate fatty acids in a collection of millet (Panicum miliaceum subsp. miliaceum) of different origins so that this information could be disseminated to breeders to advance germplasm use and breeding. To develop the calibration equations for rapid and nondestructive evaluation of fatty acid content, near-infrared reflectance spectroscopy (NIRs) spectra (1104-2494 nm) of samples ground into flour (n=100) were obtained using a dispersive spectrometer. A modified partial least-squares model was developed to predict each component. For foxtail millet germplasm, our models returned coefficients of determination (R 2 ) of 0.89, 0.89, 0.89, and 0.92 for palmitic acid, oleic acid, linoleic acid, and total fatty acids, respectively. The prediction of the external validation set (n=10) showed significant correlation between references values and NIRs values (r 2 =0.64, 0.90, 0.79, and 0.89 for palmitic acid, oleic acid, linoleic acid, and total fatty acids, respectively). Standard deviation/standard errors of cross-validation (SD/SECV) values were close to 3 (2.62, 2.40, 1.85, and 2.23 for palmitic acid, oleic acid, linoleic acid, and total fatty acids, respectively). These results indicate that these NIRs equations are functional for the mass screening and rapid quantification of the oleic and total fatty acids characterizing millet germplasm. Among the samples, IT153514 showed an especially high content of fatty acids (48.14 mg g -1 ), whereas IT123909 had a very low content (34.44 mg g -1 ).