The inheritance of corolla traits, double, hose-in-hose, and single shapes, was studied by investigating the progenies and lines from self- and cross-pollinations of Eustoma flowers. A clear segregation was observed in progenies crossed between the single and double shapes, and we identified that the hose-in-hose shape was only inherited by forming the heterozygous genotype between the native strains and/or wild species with single and double flowers. According to data on the 965 individuals from F₁ and F₂ lines, it is likely that multiple alleles control the three kinds of corolla trait. Segregation patterns of the three flower shapes could be explained by the presence of three alleles designated as D^D, D^W, and D^S in Eustoma flowers, controlling the phenotypes of double (D^DD^D and D^DD^S), hose-in-hose (D^DD^W), and single (D^SD^S, D^WD^W and D^SD^W) flowers (genotypes). Based on the results, it was concluded that the allele D^W originales from the wild Eustoma species.