Abstract Exogenous lipids serving as stimulators to improve lycopene production in Blakeslea trispora have been widely reported. However, the selection basis of exogenous lipids and their effects on intracellular lipids are not very clear. In this study, five plant oils with different fatty acid compositions were selected to investigate their effects on lycopene production, fatty acid composition and the desaturation degree of intracellular lipids. Among the oils, soybean oil, with a fatty acid composition similar to that of mycelium, exhibited the best stimulating effect on lycopene formation (improvement of 82.1%). The plant oils enhanced the total content of intracellular lipids and the desaturation degree of reserve lipids due to the alteration of fatty acid composition, especially in neutral lipids. Lycopene production was increased with the improved desaturation degree of intracellular lipids, which may be attributed to the enhancement of storage capacity for lycopene in storage lipid, thus reducing the feedback regulation of free lycopene. In addition, the increase of the desaturation degree of reserve lipids through temperature-changing fermentation also enhanced lycopene production. The present study could serve as a basis for a better understanding of the relationship between the fatty acid composition of reserve lipids and lycopene production.