ABSTRACT. ATP citrate lyase (ACLY) is the primary enzyme responsible for the synthesis of cytosolic acetyl-CoA, which is a key precursor of both fatty acid and mevalonate synthesis pathways. Genetic variation of the ACLY gene may influence multiple traits associated with animal production. Here, we identified three non-synonymous mutations in ACLY exons in five beef cattle populations using DNA pool sequencing and high-resolution melting analysis. Results from association analyses revealed that the single nucleotide polymorphism (SNP) g.17127C>T is significantly associated with chest girth (P < 0.01) and body height (P < 0.05) in the Fleckvieh x Zhangye local crossbred cattle, and with body slanting length (P < 0.05) in the Simmental x Guyuan local crossbred cattle. SNP g.40427T>C is significantly associated with an increase in chest girth (P < 0.05) in the Simmental x Huzhu cattle population. These results provide preliminary evidence that polymorphisms in the bovine ACLY gene are associated with growth traits in beef cattle in northwest China. However, a larger sample set is needed to validate these findings.