Chinese Hami melon consists of the varieties cassaba, chandalak, ameri, and zard. To show their genetic diversity, 120 melon accessions, including 24 accessions of Hami melon, were analyzed using molecular markers of nuclear and cytoplasmic genomes. All Hami melon accessions were classified as the large-seed type with seed length longer than 9 mm, like US and Spanish Inodorus melon. Conomon accessions grown in east China were all the small-seed type. Both large- and small-seed types were in landraces from Iran, Afghanistan, Pakistan, and Central Asia. Analysis of an SNP in the PS-ID region (Rpl16-Rpl14) and size polymorphism of ccSSR7 showed that the melon accessions consisted of three chloroplast genome types, that is, maternal lineages. Hami melon accessions were T/338 bp type, which differed from Spanish melon and US Honey Dew (T/333 bp type), indicating a different maternal lineage within group Inodorus. The gene diversity (D), calculated from random amplified polymorphic DNA (RAPD) and simple sequence repeat (SSR) polymorphism, was 0.476 in 120 melon accessions; the largest diversity was in Central Asian accessions (D = 0.377) but was low for Hami melon accessions (D = 0.243), even though Hami melon has diverse morphological traits, earliness, and shelf life. Reflecting such small genetic diversity, Hami melon accessions of vars. ameri and zard were grouped into cluster II, except for one accession, by the unweighted pair group method and the arithmetic mean (UPGMA) cluster analysis. Variety chandalak with distinct characters, such as early maturing and poor shelf life, was assigned to clusters IV and VI, indicating inter-varietal genetic differentiation within Hami melon. Three accessions from Turkmenistan and Afghanistan, with large seeds and T/338 bp type of chloroplast genome, were classified as cluster II with Hami melon accessions of vars. ameri and zard. We therefore concluded that Hami melon may have been transmitted from the west. The small-seed type melon of group Conomon grown in east China may have been introduced into China independently of Hami melon, because it had the A/338 bp type of the chloroplast genome and was clustered distantly from Hami melon according to nuclear genome analysis.