The Tarim Basin in Northwest China hosts petroleum reservoirs of Cambrian, Ordovician, Carboniferous, Triassic, Jurassic, Cretaceous and Tertiary ages. The sedimentary thickness in the basin reaches about 15 km and with an area of 560 000 km2, the basin is expected to contain giant oil and gas fields. It is therefore important to determine the ages and depositional environments of the petroleum source rocks. For prospective evaluation and exploration of petroleum, palynological investigations were carried out on 38 crude oil samples collected from 22 petroleum reservoirs in the Tarim Basin and on additionally 56 potential source rock samples from the same basin. In total, 173 species of spores and pollen referred to 80 genera, and 27 species of algae and fungi referred to 16 genera were identified from the non-marine Mesozoic sources. By correlating the palynormorph assemblages in the crude oil samples with those in the potential source rocks, the Triassic and Jurassic petroleum source rocks were identified. Furthermore, the palynofloras in the petroleum provide evidence for interpretation of the depositional environments of the petroleum source rocks. The affinity of the miospores indicates that the petroleum source rocks were formed in swamps in brackish to lacustrine depositional environments under warm and humid climatic conditions. The palynomorphs in the crude oils provide further information about passage and route of petroleum migration, which is significant for interpreting petroleum migration mechanisms. Additionally, the thermal alternation index (TAI) based on miospores indicates that the Triassic and Jurassic deposits in the Tarim Basin are mature petroleum source rocks.