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Adipose differentiation-related protein (ADFP) plays an important role in regulating lipid storage and affecting body fat distribution. The objective of the current research was to identify the polymorphisms in chicken ADFP gene and their associations with carcass traits. We screened all coding sequence and part of introns (totally 2155bp) of the ADFP gene by using single strand conformation polymorphism method in individuals from Sanhuang chicken, Mountainous Black-bone chicken and a commercial crossbred chicken (totally 427 birds). Three novel single-nucleotide polymorphisms were found in intron 4 (SNP1), intron 5 (SNP2) and exon 8 (SNP3), respectively. The association analyses showed that genotypes of SNP1 were significantly associated with abdominal fat weight and percentage of abdominal fat (P<0.05), genotypes of SNP2 were significantly associated with breast muscle weight and percentage of abdominal fat (P<0.05), and the main haplotype/haplotypes combination (their frequencies were higher than 5%) were marginally significantly associated with breast muscle weight, abdominal fat weight, and percentage of abdominal fat (0.05<P<0.1). Haplotype H2 (A-C-A) was an advantageous haplotype for breast muscle weight and H5 (T-C-G) was an advantageous haplotype for weight and percentage of abdominal fat. Conversely, H3H3 (A-T-G/A-T-G) was a disadvantage haplotype combination for accumulation of abdominal fat. These results suggested that polymorphisms of ADFP gene were associated with carcass traits, especially with fatness traits. ADFP is a potential major gene or in close linkage disequilibrium with the QTL affecting fatness traits in chickens.