MicroRNA-122 (miR-122), a completely conserved, liver-specific miRNA in vertebrates, is essential for the maintenance of liver homeostasis. This 22-nucleotide-length RNA regulates diverse functions such as cholesterol, glucose, and lipid metabolism as well as iron homeostasis and infection of hepatitis C virus (HCV). Landes goose, which has a good, fatty liver, has important significance for us in studying miR-122 function in goose fatty liver. In the current study, we identified miR-122 in goose liver and its expression pattern and target genes. We found that miR-122 was highly expressed in goose liver and its expression was down-regulated after overfeeding; some genes related to lipid metabolism, including prolyl 4-hydroxylase subunit alpha 1 (P4HA1); aldolase, fructose-bisphosphate B (ALDOB); and pyruvate kinase, muscle (PKM2), were predicted and validated as target genes of goose miR-122. After overexpression or inhibition of miR-122 in primary goose hepatocytes, the expression of ALDOB and PKM2 was changed, but not that of P4HA1, indicating miR-122 regulates ALDOB and PKM2 expression at the mRNA level. These findings suggest miR-122 play important roles in goose fatty liver by targeting some of the genes related to lipid metabolism.