Changes to groundwater storage (GWS) in the Great Artesian Basin (GAB) are affected by a combination of factors, including the extraction of groundwater, shifts in land use practices, and the effects of climate variability. Collectively, these factors have contributed to a decline in groundwater availability over time in some areas, impacting both the volume of groundwater reserves and the crucial replenishment of springs and surface water sources within the GAB. In the GAB, changes in GWS influence the distribution, extent, and temporal dynamics of spring wetlands and critical ecological functions they sustain. Changes to GWS, due to anthropogenic and climatic factors, can also affect the well-being of humans across Australia as they directly influence agricultural productivity and drinking water access. Reduced environmental flows and limited surface water availability are other negative effects of falling groundwater levels that could have direct or indirect impacts on humans and the productivity of ecosystems. In a changing climate, these impacts are expected to magnify progressively over time. This thesis aimed to quantify ecohydrology of the Great Artesian Basin using remote sensing technology. In this study, the term ecohydrology refers to quantifying multi-scaled relationships between hydrological and ecological processes (e.g., spring wetlands, and associated GDEs) and evaluating human impacts on the aquifer. The thesis assessed the spatio-temporal patterns of hydrological stores (groundwater and terrestrial water storage variations) and their responses to changes in climatic conditions (rainfall and evapotranspiration) using satellite observations between 2002 and 2017. Additionally, this research aimed to assess how GWS variation is related to vegetation greenness, and surface water extent under different climatic extremes (floods and droughts). This thesis also aimed to identify areas where GWS variation is particularly sensitive or vulnerable to the impacts of climate change (so-called climate hotspots) and analyzed the relationships between these hotspots and spatio-temporal changes in GWS over the GAB. [...]