This study aimed to analyze the spatial and temporal variation of the vegetation in the northern Argentine Puna, utilizing both field sampling and remote-sensing tools. The study was performed within the Pozuelos Biosphere Reserve (Jujuy province, Argentina), which aims to generate socio-economic development compatible with biodiversity conservation. Our study was designed to analyze the dynamics of the Puna vegetation at local scale and assess and monitor the seasonal (dry and wet seasons), interannual, and spatial variation of the vegetation cover, biomass, dominant species, and vegetation indices. Ten vegetation units (with differences in composition, cover, and high and low stratum biomass) were identified at our study site. The diversity of these vegetation units correlated with geomorphology and soil type. In the dry season, the vegetation unit with greatest vegetation cover and biomass was the Festuca chrysophylla grassland, whereas in the wet season, the units with greatest cover and biomass were vegas (peatlands) and short grasslands. The Festuca chrysophylla grasslands and short grasslands were located in areas with clay soils, except peatlands, associated with valleys and coarse-texture soils. The vegetation indices used (NDVI, SAVI, and MSAVI2) were able to differentiate functional types of vegetation and showed a good statistical fit with cover values. Our results suggest that the integrated utilization of remote-sensing tools and field surveys improves the assessment of the Puna vegetation and would allow a periodic monitoring at production unit scale taking into account its spatial and temporal variation.