The aim of this study was to detect the polymorphisms in the promoter region of PGR gene based on PCR-Restriction Fragment Length Polymorphism technique (PCR-RFLP). Genomic DNA were extracted and amplified from 100 animal belonging to four rabbit populations; an Egyptian synthetic rabbit line named Moshtohor line (M-line) and their parents of Spanish V-line and Sinai Gabali rabbit breed, in addition to using the French Giant Papillon (FGP) as a reference population. PCR-RFLP was applied on 558 bp using the Eco31I restriction endonuclease. PCR-RFLP for PGR gene revealed three genotypes of GG, AA and GA, and the genotype of GG yielding two bands of 416 and 142 bp, the genotype AA yielding a single 558 bp band, and the heterozygote genotype GA yielding all the three bands of 558, 416 and 142 bp. The genotypic frequency of GA ranged from 0.680 in V-line to 0.880 in FGP and was higher than AA and GG genotypes in all populations studied. The frequency of A allele was higher than the G allele in all populations except in M-line where G allele recorded the highest allele frequency of 0.540. The highest effective number of alleles (Ne) for SNP of PGR gene was recorded for M-line (1.987), followed by FGP (1.972), while the lowest number was recorded for V-line (1.891). The four populations studied were not in Hardy-Weinberg equilibrium (P< 0.05 or P< 0.001). The observed (Ho), expected (He) heterozygosity and the polymorphic information content (PIC) averaged 1.943, 0.485 and 0.367, respectively, i.e. the four rabbit populations showed intermediate levels of genetic diversity.