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[Objectives] This study aimed to verify the correlation between the c.-751 A>C in ACSF2(encoded by acyl-CoA synthetase family member 2) promoter and laying performance of Yangzhou goose, and analyze the effect of the SNP on ACSF2 expression. [Methods] We used allele specific PCR(AS-PCR) to analyze the genotype of SNP c.-751 A>C in a 343 Yangzhou geese population, and performed the correlation analysis between genotype and laying performance of Yangzhou goose. Quantitative real-time PCR(qPCR) and luciferase reporter gene vector were used to detect the difference between AA and CC promoter activity. ACSF2 over-expression was performed to analyze its role in the energy metabolism of granulosa cells and laying performance. [Results] Genotyping and association analysis results showed that the egg numbers of geese with AA genotype were significantly higher than those CC ones. Quantitative real-time PCR analysis indicated that AA individuals showed lower ACSF2 expression level in ovary tissues. Moreover, the overexpression experiment indicated that ACSF2 was involved in the energy metabolism of granulosa cells. [Conclusions] We identified a functional mutation in the promoter region of ACSF2 affecting gene expression in Yangzhou goose, and indicated the role of ACSF2 in laying performance by regulating energy metabolism of granulosa cells. Our results provide insight into the improvement of goose laying performance by ACSF2 genetic variation selection. [ABSTRACT FROM AUTHOR]