Abstract: Twenty-six species (or forms) of fossil pollen and spores referred to 24 genera extracted from the crude oil samples collected from the Yumen oil-bearing region in West Gansu are observed and the definition of the petroleum sporo-pollen assemblage is given by the authors. On the basis of palynological investigation, the oil source rock of this oil-bearing region is tentatively discussed in this paper. The fossil pollen and spores found from the Tertiary petroleum in this region may be divided into two groups, because they are extremely different in fossilized state. The group of spores and pollen with higher fossilized state is mainly consisted of Schizaeoisporites zizyphinus, Gibotiumspora paradoxa, Monosulcites minimus and Bennettiteaepollenites sp. The other group comprises Pinus, Picea, Ephedra, Potamogeton, Lilium, Ulmus, Chenopodium, Nymphaea, Eutrema, Lens, Nitraria, Tamarix, Achillea, Artemisia, Solidago, Bidens, Graminidites, Cypheraceaepollis, etc. The above mentioned petroleum sporo-pollen assemblage of the tertiary oil reservoir rock is a gathering of fossil pollen and spores brought from the Lower Cretaceous deposits and the tertiary deposits respectively. The appearance of the Early Cretaceous pollen and spores bearing in the black shales of the Lower Xinminbu Formation in the tertiary petroleum may be considered to be a result of petroleum migration. And it reflects that the oil source rock might belong to Early Cretaceous Series. The fossil pollen and spores extracted from the Early Cretaceous petroleum in this region contain merely Schizaeoisporites zizyphinus, Monosulcites minimus, Bennettiteaepollenites sp., Psophosphaera, etc. The tertiary pollen and spores are completely absent there. So it indicates that both the oil source rock and the oil reservoir rock should belong to the fossil-bearing Lower Xinminbu Formation. In addition, there are some pollen of Monosulcites and Psophosphaera in the petroleum reserved in the fissures of the Silurian metamorphic rock. They are also probably brought from the Lower Xinminbu Formation by oil and gas in the process of their migration. In any case, judging from the pollen and spores found from the crude oil, the oil source rock of this oil-bearing region is deduced to be the black shales of the Lower Xinminbu Formation referred to as Early Cretaceous Epoch.