The nutritional diagnosis of crops is carried out through costly elemental analyses of different plant organs, particularly leaves, in the laboratory. However, visible and near-infrared (Vis-NIR) spectroscopy of unprocessed plant samples has a high potential as a faster, non-destructive, environmental-friendly alternative to elemental analyses. In this work, the potential of this technique to estimate the concentrations of macronutrients such as nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), and magnesium (Mg), and micronutrients such as iron (Fe), manganese (Mn) and boron (B), in persimmon (Diospyros kaki L.) ‘Rojo Brillante’ leaves, has been investigated. Throughout the crop cycle variable rates of N and K were applied to obtain six nutritional status levels in persimmon trees in an experimental orchard. Then, leaves were systematically sampled throughout the cropping season from the different nutritional levels and spectral reflectance measurements were acquired in the 430–1040 nm wavelength range. The concentrations of nutrients were determined by inductively coupled plasma optical emission spectrometry (ICP-OES) for P, K, Ca, Mg, Fe, Mn and B after microwave digestion, while the Kjeldahl method was used for N. Then, partial least squares regression (PLS-R) was used to model the concentrations of these nutrients from the reflectance measurements of the leaves. The model was calibrated using 75% of the samples while the remaining 25% were left as the independent test set for external validation. The results of the test set indicated an acceptable validation for most of the nutrients, with determination coefficients (R2) of 0.74 for N and P, 0.54 for K, 0.77 for Ca, 0.60 for Mg, 0.39 for Fe, 0.69 for Mn and 0.83 for B. These findings support the potential use of Vis-NIR spectrometric techniques as an alternative to conventional laboratory methods for the persimmon nutritional status diagnosis although more research is needed to know how the models developed one year perform in ensuing years.