Nineteen polymorphic and eleven monomorphic isozyme loci were identified in thirteen enzyme systems in a survey of four-hundred melon (Cucumis melo L.) accessions. Segregation of allozymes in F2 and backcross (BC) families for isozyme loci agreed with the expected 1:2:1 and 1:1 segregation ratios (P <0.01). Eleven isozyme loci were linked and were integrated to form a map containing two linkage groups spanning 98 cM with a mean linkage distance of ≈9 cM. Linkage groups (A and B) contain the following loci in the order: A Fdp-2, Pgd, Pgm, Mpi-1, Idh, and Ac, and B Pep-gl, Mdh-2, Mdh-4, Mdh-5, Mdh-6. The remaining eight loci (Acp-1, Acp-4, Ak-4, Fdp-1, Gpi, Mpi-2, Pep-la, and Pep-pap) segregated independently. The isozyme map constructed in this study provides genomic information for future linkage studies with economically important traits and concensus map construction through map merging.