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Miscanthus × giganteus (Miscanthus) is a warm-season perennial grass grown for bioenergy feedstock production. Nitrogen (N) fertilizer management is crucial for the sustainability of Miscanthus production. In our two-year study (2018 and 2019), we investigated the role of vegetation indices (VIs) in evaluating N fertilization (0 N, 56 N, 112 N, and 168 N kg ha−1) impacts on Miscanthus biomass yield and stand health. The flight campaigns were conducted early, middle, and late during the summer growing season. Among the VIs, mid-summer growing season NDRE provided the best prediction of fresh biomass (R2 = 0.87 and 0.97) and dry biomass (R2 = 0.89 and 0.97) in 2018 and 2019, respectively. The VIs generally showed that it was possible to distinguish between 0 N and 168 N treatments, but neither 0 N and 56 N kg ha−1 nor 112 N and 168 N kg ha−1 could be separated. The results from this study highlight the importance of moderate application of N (112 kg N ha−1) in improving and maintaining the stand health and biomass yield of Miscanthus over time and suggest that mid-summer growing season VIs, NDRE in particular, can be useful for assessment of Miscanthus stand health and biomass yield. [ABSTRACT FROM AUTHOR]