Furthermore, TILLinG produces a broad range of mutations including nonsense, missense and splicing mutations which can be used for protein domain annotation. This technique has been successfully applied to a large variety of organisms including plants and animals and has become the method of choice for gene functional analysis in crop species. In cucumber, sex determination is genetically governed by the genes Monoecious, gynoecious and androecious, and the interplay of these three genes can result in a range of sexual phenotypes. Monoecious and andromonoecious individuals bear male flowers and LDN-193189 respectively female or hermaphrodite flowers. Gynoecious and hermaphrodite individuals only bear female and hermaphrodite flowers, respectively. The androecious gene increases maleness and plants of the aaff genotype are androecious bearing only male flowers. In melon, sex determination is genetically governed by the genes andromonoecious and gynoecious and plants of,LEE011 and genotypes are monoecious, andromonoe-cious, gynoecious and hermaphrodite, respectively. Phenotypical-ly, melon andromonoecious gene appears to act similarly to cucumber Monoecious gene. In both species, the dominant allele, M in cucumber and A in melon, suppresses stamen development in pistillated flowers without affecting male flower formation, whereas the recessive allele, m in cucumber and a in melon ‘‘releases’’ such inhibition, resulting in bisexual flowers instead of female flowers. In both Cucumis species, sexual morphs can be also modified by hormonal and environmental factors, with ethylene playing a major role. Consistent with ethylene being a feminizing agent, we previously demonstrated that the melon A gene encodes for the rate-limiting enzyme in ethylene biosynthesis, the 1-aminocyclopropane-1-carboxylic acid synthase, CmACS-7. CmACS-7 is also specifically expressed in carpel primordia, and in andromonoecious genotypes, a missense mutation leads to loss of enzymatic activity. In cucumber, using a genetic approach we showed that CsACS2, a cucumber ACS highly homologous to CmACS-7, co-segregates with the Monoecious locus.