Abstract:The present work assessed the usefulness of a set of spectral indices obtained from an unmanned aerial system (UAS) for tracking spatial and temporal variability of nitrogen (N) status as well as for predicting lint yield in a commercial cotton (Gossypium hirsutum L.) farm. Organic, inorganic and a combination of both types of fertilizers were used to provide a range of eight N rates from 0 to 340 kg N ha −1 . Multi-spectral images (reflectance in the blue, green, red, red edge and near infrared bands) were acquired on seven days throughout the season, from 62 to 169 days after sowing (DAS), and data were used to compute structure-and chlorophyll-sensitive vegetation indices (VIs). Above-ground plant biomass was sampled at first flower, first cracked boll and maturity and total plant N concentration (N%) and N uptake determined. Lint yield was determined at harvest and the relationships with the VIs explored. Results showed that differences in plant N% and N uptake between treatments increased as the season progressed. Early in the season, when fertilizer applications can still have an effect on lint yield, the simplified canopy chlorophyll content index (SCCCI) was the index that best explained the variation in N uptake and plant N% between treatments. Around first cracked boll and maturity, the linear regression obtained for the relationships between the VIs and both plant N% and N uptake was statistically significant, with the highest r 2 values obtained at maturity. The normalized difference red edge (NDRE) index, and SCCCI were generally the indices that best distinguished the treatments according to the N uptake and total plant N%. Treatments with the highest N rates (from 307 to 340 kg N ha −1 ) had lower normalized difference vegetation index (NDVI) than treatments with 0 and 130 kg N ha −1 at the first measurement day (62 DAS), suggesting that factors other than fertilization N rate affected plant growth at this early stage of the crop. This fact affected the earliest date at which the structure-sensitive indices NDVI and the visible atmospherically resistant index (VARI) enabled yield prediction (97 DAS). A statistically significant linear regression was obtained for the relationships between SCCCI and NDRE with lint yield at 83 DAS. Overall, this study shows the practicality of using an UAS to monitor the spatial and temporal variability of cotton N status in commercial farms. It also illustrates the challenges of using multi-spectral information for fertilization recommendation in cotton at early stages of the crop.