Members of the Cucurbitaceae family display a range of sexual phenotypes including various combinations of male, female, or bisexual flowers. Ethylene appears to be a key hormone regulating the sex determination process. Application of ethylene, or inhibition of ethylene action, increases or decreases the number of pistil-bearing buds, respectively. Elevated levels of ethylene production and expression of genes for ethylene biosynthesis, have been correlated with pistillate flower production. In this study, we sought to determine the effect of modified endogenous ethylene production on sex expression by constitutively expressing ACS (1-aminocyclopropane-1-carboxylate synthase), the first committed enzyme for ethylene biosynthesis, in transgenic melons (Cucumis melo L.). Most melon genotypes are andromonoecious, where an initial phase of male flowers is followed by a mixture of bisexual and male flowers. ACS melon plants showed increased ethylene production by leaves and flower buds, and increased femaleness as measured by earlier and increased number of bisexual buds. ACS melons also had earlier and increased number of bisexual buds that matured to anthesis, suggesting that ethylene is important not only for sex determination, but also for development of the bisexual bud to maturity. Field studies showed that ACS melons had earlier mature bisexual flowers, earlier fruit set, and increased number of fruit set on closely spaced nodes on the main stem. These results provide a direct demonstration of the importance of endogenous ethylene production for female reproductive processes in melon.

Sexual diversity expressed by Curcurbitaceae species is a primary example of developmental plasticity in plants. Ethylene, which promotes femaleness (carpel development), plays a key role in sex determination. We sought to determine the critical location for ethylene perception in developing floral primodia. The dominant negative Arabidopsis ethylene response mutant gene, etr1-1, was introduced into melon (Cucumis melo L.) plants under control of the constitutive cauliflower mosaic virus (CaMV) 35S promoter, or floral-targeted Apetela3 (AP3) and Crab's Claw (CRC) promoters, which in Arabidopsis, promote expression in petal and stamen, and carpel and nectary primordia, respectively. Based on effects of exogenous ethylene, it was predicted that inhibition of ethylene perception by carpel primordia would inhibit carpel development. Constitutive expression of etr1-1 caused several phenotypes associated with ethylene insensitivity, verifying that etr1-1 inhibits ethylene perception in the heterologous melon system. Carpel-bearing bud production was essentially abolished in 35S::etr1-1 melons, providing direct demonstration of the requirement for ethylene perception for carpel development. CRC::etr1-1 plants, however, showed enhanced femaleness as manifested by earlier and increased number of carpel-bearing buds, and production of female (rather than bisexual) buds. Despite increased carpel-bearing bud formation, a greater proportion of the CRC::etr1-1 carpel-bearing buds aborted before anthesis. AP3::etr1-1 plants showed increased maleness by nearly exclusive staminate flower production, and poorly developed carpels in the rare bisexual flowers. These results indicate that ethylene perception by the stamen (or petal) primordia plays a critical role in promoting carpel development at the time of sex determination, while ethylene perception by the carpel is important for maturation of carpel-bearing flowers to anthesis.

The Cucurbitaceae family is noted for a diversity of sex expression phenotypes. Typically, a phase of male flowers precedes either female or bisexual flower production. Sex determination of individual flowers is regulated by a combination of genetic, environmental, and hormonal factors. Ethylene, auxins, and gibberellins have all been shown to influence flower sex expression in cucurbits. Ethylene, which promotes femaleness, plays a predominant role. In this study, we tested whether brassinosteroids (BR), a more recently identified class of plant hormones, also influences cucurbit sex expression. Applied epi-brassinolide (epi-BL) caused a significant decrease in time of appearance of the first female flower on monoecious cucumber plants, and increased total female flowers on the main stem. Increasing concentrations had a stronger effect. Of the three species tested, cucumber, melon and zucchini, cucumber was the most responsive to BR. Application of epi-BL also caused an increase in ethylene production by cucumber and zucchini seedlings, suggesting that the BR effect may be mediated by ethylene. To investigate the possible relationship between BR and ethylene on sex expression, we identified the concentration of ethephon (5 ppm) that caused an increase in ethylene production comparable to that induced by 10 μm epi-BL (approximately two-fold). Treatment with 5 ppm ethephon was sufficient to increase femaleness of cucumber plants, but not zucchini plants, suggesting that the difference in response to epi-BL treatment may reflect differences in sensitivity to ethylene. Collectively, our results indicate that application of brassinosteroids to cucumber cause earlier and increased female flower production, and that the effects may be mediated, at least in part, by brassinosteroid-induced production of ethylene.

The cucurbit family is noted for diversity in sex expression phenotypes. Typically, a phase of male flowers precedes the appearance of female or hermaphrodite flowers. Sex determination of individual flowers is regulated by genetic, environmental, and hormonal factors. Ethylene, auxins, and gibberellins all influence flower sex, with ethylene, which promotes femaleness, playing a predominant role. In this study, we tested whether brassinosteroids, a more recently identified class of plant hormones, also influence cucurbit sex expression. Applied epi-brassinolide (epi-BL) caused a significant decrease in time of appearance of the first female flower on monoecious cucumber plants, and increased total female flowers on the main stem. Increasing concentrations had a stronger effect. Of the three species tested, cucumber, melon, and zucchini, cucumber was the most responsive. Application of epi-BL also caused an increase in ethylene production by cucumber and zucchini seedlings, suggesting that the BR effect may be mediated by ethylene. To investigate the possible relationship between BR and ethylene on sex expression, we identified the concentration of ethephon (5 ppm) that caused an increase in ethylene production comparable to that induced by 10 μm epi-BL (about two-fold). Treatment with 5 ppm ethephon was sufficient to increase femaleness of cucumber plants, but not zucchini plants, suggesting that the difference in response to epi-BL may reflect differences in sensitivity to ethylene. Collectively, our results indicate that application of brassinosteroids to cucumber cause earlier and increased female flower production, and that the effects may be mediated, at least in part, by brassinosteroid-induced increased production of ethylene.