Understanding the evolution of sex determination in plants requires the cloning and the characterization of sex determination genes. Monoecy is characterized by the presence of both male and female flowers on the same plant. Andromonoecy is characterized by plants carrying both male and bisexual flowers. In watermelon, the transition between these two sexual forms is controlled by the identity of the alleles at the A locus. We previously showed, in two Cucumis species, melon and cucumber, that the transition from monoecy to andromonoecy results from mutations in 1-aminocyclopropane-1-carboxylic acid synthase (ACS) gene, ACS-7/ACS2. To test whether the ACS-7/ACS2 function is conserved in cucurbits, we cloned and characterized ClACS7 in watermelon. We demonstrated co-segregation of ClACS7, the homolog of CmACS-7/CsACS2, with the A locus. Sequence analysis of ClACS7 in watermelon accessions identified three ClACS7 isoforms, two in andromonoecious and one in monoecious lines. To determine whether the andromonoecious phenotype is due to a loss of ACS enzymatic activity, we expressed and assayed the activity of the three protein isoforms. Like in melon and cucumber, the isoforms from the andromonoecious lines showed reduced to no enzymatic activity and the isoform from the monoecious line was active. Consistent with this, the mutations leading andromonoecy were clustered in the active site of the enzyme. Based on this, we concluded that active ClACS7 enzyme leads to the development of female flowers in monoecious lines, whereas a reduction of enzymatic activity yields hermaphrodite flowers. ClACS7, like CmACS-7/CsACS2 in melon and cucumber, is highly expressed in carpel primordia of buds determined to develop carpels and not in male flowers. Based on this finding and previous investigations, we concluded that the monoecy gene, ACS7, likely predated the separation of the Cucumis and Citrullus genera.

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: A total of 67 watermelon accessions which include 37 accessions from Korean and 27 accessions from Turkish germplasm and 3 accessions of other related species from USA were investigated for morphological characteristics. The UPOV descriptor list for 56 characters (6 seedlings, 4 plants, 11 leaves, 5 flowers, 23 fruits and 7 seeds) was used in characterization. In addition, eight quantitative characters, hypocotyl length, cotyledon width, cotyledon length, fruit weight, fruit length, fruit width, thickness of outer layer of pericarp and soluble solid content were also measured. The 56 qualitatively scored characters were analyzed by principle coordinate analysis (PCoA) while the eight quantitative ones were subjected to principle component analysis (PCA). Morphological characterization result demonstrated that the accessions displayed high morphological diversity(how much percent?). A high level of phenotypic diversity was observed from the results of morphological characterization. However, plant growth habit and leaf blade flecking showed constant characters for all of the accessions. The Korean and Turkish watermelon genotypes are diverse groups and can be separated by both multivariate analysis of morphological characters although the grouping was more apparent in PCoS results.

In this study, the Turkish watermelon genetic resources were collected in 2004 and morphologically characterized in 2005. The collection comprising of traditional cultivars and local accessions were collected from 15 different provinces of Southeastern Anatolia, Aegean, Marmara-Thrace, Middle Anatolia and Mediterranean regions of Turkey where local watermelon types are widely grown. A total of 134 accessions were characterized morphologically according to UPOV descriptor list for 56 qualitative characters (6 seedling, 4 plant, 11 leaf, 5 flower, 23 fruit and 7 seed) and morphologic relatedness were examined. Additionally, 22 quantitative characters were also measured. Our results indicated that the accessions studied have a great deal of variation for most of the morphological traits. The accessions were divided into five different groups according to principle component analysis. However, the geographical origin was not a great source for grouping.

Genetic diversity of the Turkish watermelon genetic resources was evaluated using different Citrullus species, wild relatives, foreign landraces, open pollinated (OP) and commercial hybrid cultivars by RAPD markers. The germplasm was consisted of 303 accessions collected from various geographical regions. Twenty-two of 35 RAPD primers generated a total of 241 reproducible bands, 146 (60.6%) of which were polymorphic. Based on the RAPD data the genetic similarity coefficients were calculated and the dendrogram was constructed using UPGMA (Unweighted pair-group method with arithmetic average). Cluster analysis of the 303 accessions employing RAPD data resulted in a multi-branched dendrogram indicating that most of the Turkish accessions belonging to var. lanatus of Citrullus lanatus (Thunb.) Matsum et Nakai were grouped together. Accessions of different Citrullus species and Praecitrullus fistulosus (Stocks) Pangalo formed distant clusters from C. lanatus var. lanatus. Among 303 accessions, a subset of 56 accessions was selected representing different groups and a second dendrogram was constructed. The genetic similarity coefficients (GS) within the Turkish accessions were ranged from 0.76 to 1.00 with 0.94 average indicating that they are closely related. Taken together, our results indicated that low genetic variability exist among the watermelon genetic resources collected from Turkey contrary to their remarkable phenotypic diversity.