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We studied the effect of dietary linseed oil (LSO) supplementation and DGAT1 K232A (DGAT1) polymorphism on the triacylglycerol composition and crystallization of bovine milk fat. LSO supplementation increased unsaturated triacylglycerols, notably in the C52-C54 carbon range, while reducing the saturated C29-C49 triacylglycerols. These changes were associated with an increase in the low-melting fraction and the crystal lamellar thickness, as well as a reduction in the medium and high-melting fractions and the formation of the most abundant crystal type at 20 °C (β'-2 polymorph). Furthermore, DGAT1 KK was associated with higher levels of odd-chain saturated triacylglycerols than DGAT1 AA, and it was also associated with an increase in the high-melting fraction and the endset melting temperature. An interaction between diet and DGAT1 for the unsaturated C54 triacylglycerols accentuated the effects of LSO supplementation with DGAT1 AA. These findings show that genetic polymorphism and cows' diet can have considerable effects on milk fat properties., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022. Published by Elsevier Ltd.)