In this paper, the spatio-temporal variation and attribution of land surface temperature in Bosten Lake Basin were studied by methods of spatial analysis, geo-statistical and trend analysis using LST product of MODIS, LUCC data and observed LST data from 2001 to 2014. The results indicated that: (1) MODIS LST product was enough to apply for the analysis of spatio-temporal distribution characteristics of land surface temperature in Bosten Lake Basin(mean R2 =0.96); (2) Inter-annual variation of LST was obvious, especially in the year of 2002, 2007 and 2013, and exceeded the multiannual mean LST value by 0.45℃, 0.59℃ and 0.29℃ respectively. Furthermore, the seasonal variation of LST showed a unimodel pattern, and its peak value occured in July. (3) The LST variation of southeast was higher than the northwest areas, especially in the low vegetation coverage areas. The spatial distribution of LST was related to land cover types,and the order of LST intensity by land cover types was desert > bare soil > dry land > sparse grassland > lake > grass land > herbaceous swamp > meadow; (4) According to the LST variation characteristics of Bosten Lake Basin, the LST trends of severely reduced, slightly reduced, basically unchanged, slightly increased and significantly increased, and their area accounted for 5%,13%,39%,36% and 7% respectively. The spatio-temporal variation of LST not only was influenced by the global climate change, but also effected by temperature field of different LUCC in different district and zone. Vegetation cover and its vertical distribution contributed to the spatio-temporal distribution of regional LST.