The isotopic composition of water vapor can be used to track atmospheric hydrological processes and to evaluate numerical models simulating the water cycle. Accurate model–observation comparisons require understanding the spatial and temporal variability of tropospheric water vapor isotopes. The challenging task of obtaining highly resolved water vapor isotopic observations is typically addressed through airborne measurements performed aboard conventional aircraft, but these offer limited microscale insights. This study uses ultralight aircraft observations to investigate water vapor isotopic composition in the lower troposphere over southern France in late summer 2021. Combining observations with models, we identify key drivers of isotopic variability and detect short-lived, small-scale processes. The key findings of this study are that (i) at hourly and sub-daily scales, vertical mixing is the primary driver of isotopic variability in the lowermost troposphere above the study site; (ii) evapotranspiration significantly impacts the boundary layer water vapor isotopic signature, as revealed by the δ18O–δD relationship; and (iii) while water vapor isotopes generally follow large-scale humidity patterns, with separation distances that might range up to 100–300 km, they also reveal distinct small-scale structures (approximately hundreds of meters) that are not fully explained by humidity variations alone, highlighting sensitivity of water vapor isotopic composition to additional fine-scale processes. The latter are particularly evident for δD, which also exhibit the largest differences in horizontal and vertical gradients. Combined with other airborne datasets, our results support a simple model driven by surface observations to simulate tropospheric δD vertical profiles, improving surface–satellite comparisons.