In plants, unisexual flowers derived from developmental sex determination form separate stamens and pistils that facilitate cross pollination. In cucumber and melon, ethylene plays a key role in sex determination. Six sex determination-related genes have been identified in ethylene biosynthesis in these Cucumis species. The interactions among these genes are thought to involve ethylene signaling; however, the underlying mechanism of regulation remains unknown. In this study, hormone treatment and qPCR assays were used to confirm expression of these sex determination-related genes in cucumber and melon is ethylene sensitive. RNA-Seq analysis subsequently helped identify the ethylene responsive factor (ERF) gene, CsERF110, related to ethylene signaling and sex determination. CsERF110 and its melon ortholog, CmERF110, shared a conserved AP2/ERF domain and showed ethylene-sensitive expression. Yeast one-hybrid and ChIP-PCR assays further indicated that CsERF110 bound to at least two sites in the promoter fragment of CsACS11, while transient transformation analysis showed that CsERF110 and CmERF110 enhance CsACS11 and CmACS11 promoter activity, respectively. Taken together, these findings suggest that CsERF110 and CmERF110 respond to ethylene signaling, mediating ethylene-regulated transcription of CsACS11 and CmACS11 in cucumber and melon, respectively. Furthermore, the mechanism involved in its regulation is thought to be conserved in these two Cucumis species.

A novel dwarf cucumber mutant, scp-2, displays a typical BR biosynthesis-deficient phenotype, which is due to a mutation in CsDET2 for a steroid 5-alpha-reductase. Brassinosteroids (BRs) are a group of plant hormones that play important roles in the development of plant architecture, and extreme dwarfism is a typical outcome of BR-deficiency. Most cucumber (Cucumis sativus L.) varieties have an indeterminate growth habit, and dwarfism may have its value in manipulation of plant architecture and improve production in certain production systems. In this study, we identified a spontaneous dwarf mutant, super compact-2 (scp-2), that also has dark green, wrinkle leaves. Genetic analyses indicated that scp-2 was different from two previously reported dwarf mutants: compact (cp) and super compact-1 (scp-1). Map-based cloning revealed that the mutant phenotype was due to two single nucleotide polymorphism and a single-base insertion in the CsDET2 gene that resulted in a missense mutation in a conserved amino acid and thus a truncated protein lacking the conserved catalytic domains in the predicted steroid 5α-reductase protein. Measurement of endogenous hormone levels indicated a reduced level of brassinolide (BL, a bioactive BR) in scp-2, and the mutant phenotype could be partially rescued by the application of epibrassinolide (EBR). In addition, scp-2 mutant seedlings exhibited dark-grown de-etiolation, and defects in cell elongation and vascular development. These data support that scp-2 is a BR biosynthesis-deficient mutant, and that the CsDET2 gene plays a key role in BR biosynthesis in cucumber. We also described the systemic BR responses and discussed the specific BR-related phenotypes in cucumber plants.

The family Cucurbitaceae includes many economically important crops, such as cucumber (Cucumis sativus), melon (Cucumis melo), watermelon (Citrullus lanatus), and zucchini (Cucurbita pepo), which share homologous gene pathways that control similar phenotypes. Sex determination is a research hotspot associated with yield and quality, and the genes involved are highly orthologous and conserved in cucurbits. In the field, six normal sex types have been categorized according to the distribution of female, male, or bisexual flowers in a given plant. To date, five orthologous genes involved in sex determination have been cloned, and their various combinations and expression patterns can explain all the identified sex types. In addition to genetic mechanisms, ethylene controls sex expression in this family. Two ethylene signaling components have been identified recently, which will help us to explore the ethylene signaling-mediated interactions among sex-related genes. This review discusses recent advances relating to the mechanism of sex determination in cucurbits and the prospects for research in this area.

The WUSCHEL-related homeobox1 (WOX1) transcription factor plays an important role in lateral growth of plant organs; however, the underlying mechanisms in the regulation of reproductive development are largely unknown. Cucumber (Cucumis sativus) has separate male and female flowers, facilitating the study of the role of WOX1 in stamen and carpel development. Here, we identified a mango fruit (mf) mutant in cucumber, which displayed multiple defects in flower growth as well as male and female sterility. Map-based cloning showed that Mf encodes a WOX1-type transcriptional regulator (CsWOX1), and that the mf mutant encodes a truncated protein lacking the conserved WUS box. Further analysis showed that elevated expression of CsWOX1 was responsible for the mutant phenotype in cucumber and Arabidopsis. Comparative transcriptome profiling revealed certain key players and CsWOX1-associated networks that regulate reproductive development. CsWOX1 directly interacts with cucumber SPOROCYTELESS (CsSPL), and many genes in the CsSPL-mediated pathway were down-regulated in plants with the mutant allele at the Mf locus. In addition, auxin distribution was affected in both male and female flowers of the mutant. Taking together, these data suggest that CsWOX1 may regulate early reproductive organ development and be involved in sporogenesis via the CsSPL-mediated pathway and/or modulate auxin signaling in cucumber.

A 14 bp deletion in CsACS2 gene encoding a truncated loss-of-function protein is responsible for elongated fruit shape and perfect flowers in cucumber. In cucumber (Cucumis sativus L.), sex expression and fruit shape are important components of biological and marketable yield. The association of fruit shape and sex expression is a very interesting phenomenon. The sex determination is controlled primarily by the F (female) and M (monoecy) loci. Homozygous recessive mm plants bear bisexual (perfect) flowers, and the fruits are often round shaped. CsACS2 encoding the 1-aminocyclopropane-1-carboxylic acid synthase has been shown to be the candidate gene for the m locus. We recently identified an andromonoecious cucumber line H38 that has bisexual flowers but elongated fruits. To rapidly clone this monoecious gene in H38, we developed a tri-parent mapping strategy, which took advantage of the high-density Gy14 × 9930 cucumber genetic map and the powder of bulk segregant analysis. Microsatellite markers from the Gy14 × 9930 map were used to screen two pairs of unisexual and bisexual bulks constructed from H38 × Gy14 and H38 × 9930 F2 populations. Polymorphic markers were identified and used to quickly develop a framework map and place the monoecious locus of H38 in cucumber chromosome 1. Further fine mapping allowed identification of a novel allele, m-1, at the monoecious locus to control the bisexual flower in H38, which was due to a 14 bp deletion in the third exon of the CsACS2 gene encoding a truncated loss-of-function protein of the cucumber 1-aminocyclopropane-1-carboxylic acid synthase. This new allele provides a valuable tool in understanding the molecular mechanisms of CsACS2 in the relationships of sex determination, fruit shape, and CsACS activities in cucumber.