The invasion of alien species is a significant threat to global biodiversity and the top driver of climate change. The present study was conducted in the Great Rann of Kachchh, part of Kachchh Biosphere Reserve, Gujarat, India, which has been severely affected by invasion of Prosopis juliflora. The invasive weed infestation has been identified using multi-temporal remote sensing datasets of 1977, 1990, 1999, 2005 and 2011. Spatial analyses of the transition matrix, extent of invasive colonies, patchiness, coalescence and rate of spread were carried out. During the study period of three and half decades, almost 295 km2 of the natural land cover was converted into Prosopis cover. This study has shown an increment of 42.9% of area under Prosopis cover in the Great Rann of Kachchh, part of the Kachchh Biosphere Reserve during 1977 to 2011. Spatial analysis indicates high occupancy of Prosopis cover with most of the invasion (95.9%) occurring in the grasslands and only 4.1% in other land cover types. The process of Prosopis invasion shows high patch initiation, followed by coalescence, indicating aggressive colonization of species. The number of patches within an area of > 1 km2 increased from 1977 to 2011, indicating the formation of new Prosopis habitats by replacing the grasslands. The largest patch of Prosopis cover increased from 144 km2 in 1977 to 430 km2 in 2011. The estimated mean patch size was 7.8 km2 in 1977. The mean patch size was largest during 2011, i.e., 9 km2. The annual spread rate for Prosopis has been estimated as 2.1% during 2005–2011. The present work has investigated the long term changes in Prosopis cover in the Great Rann of Kachchh, part of Kachchh Biosphere Reserve. The spatial database generated will be useful in preparing strategies for the management of Prosopis juliflora.