Unmanned aerial vehicle time‐series phenotyping illuminates genotype‐specific responses to off‐target dicamba exposure in soybean

Abstract Dicamba, a synthetic auxin herbicide, is widely used for postemergence weed control. However, the high potential for volatility and spray drift raises concerns about damage to nontarget dicot crops lacking the dicamba monooxygenase transgene, which provides bioengineered resistance. Our objective was to evaluate the practical application of unmanned aerial vehicle (UAV)‐based multispectral imaging for quantifying off‐target dicamba injury, using non‐genetically modified (GM) soybean ( Glycine max (L.) Merr.) genotypes with known tolerant (T) and sensitive (S) responses to provide a reference framework. We conducted field experiments in 2021 and 2022 using varying dicamba rates. Temporal visual injury estimates (0–5 scale) were recorded approximately every 5 days for 6–8 weeks. Significant differences were detected between T and S lines, especially with 1/500 single and weekly repeated treatments, which provided the largest separation. To augment the precision of visual injury assessment, UAV‐based multispectral imagery from each of multiple flights was used to calculate four vegetation indices (VIs): green leaf index (GLI), normalized excess green index (ExG2), normalized green red difference index (NGRDI), and red edge normalized difference vegetation index (RENDVI). These VIs captured differences between T and S lines, indicating dicamba injury can be quantified through remote sensing across contrasting soybean genotypes. Strong negative correlations were observed between visual injury scores and VI scores across treatments at each flight date, with overall correlations for GLI, ExG2, NGRDI, and RENDVI being −0.43, −0.43, −0.44, and −0.39 in 2021 and −0.54, −0.54, −0.64, and −0.57 in 2022. These results support the utility of UAV‐derived VIs for detecting and quantifying off‐target herbicide injury in soybean, facilitating scientific research efforts to develop non‐GM‐tolerant genotypes.