Two androecious mutations reveal the crucial role of ethylene receptors in the initiation of female flower development in Cucurbita pepo

Ethylene is the key regulator of sex determination in monoecious species of the family Cucurbitaceae. This hormone determines which individual floral meristems develop as female or male flowers and the female flowering transition. The sex determination genes discovered so far code for ethylene biosynthesis enzymes, but little is known about the importance of ethylene signaling components. In this paper we characterize two novel ethylene‐insensitive mutations (etr1a‐1 and etr1b) which block the female flowering transition of Cucurbita pepo; this makes plants produce male flowers indefinitely (androecy). Two missense mutations in the ethylene‐binding domain of the ethylene receptors CpETR1A or CpETR1B were identified as the causal mutations of these phenotypes by using whole‐genome resequencing. The distinctive phenotypes of single and double mutants for four etr mutations have demonstrated that the final level of ethylene insensitivity depends upon the strength and dosage of mutant alleles for at least three cooperating ETR genes, and that the level of ethylene insensitivity determines the final sex phenotype of the plant. The sex phenotype ranges from monoecy in ethylene‐sensitive wild‐type plants to androecy in the strongest ethylene‐insensitive ones, via andromonoecy in partially ethylene‐insensitive plants. The induction of female flowering transition was found to be associated with upregulation of CpACS11, CpACO2 and CpACS27, three ethylene biosynthesis genes required for female flower development. A model is proposed herein, integrating both ethylene biosynthesis and receptor genes into the genetic network which regulates sex determination in C. pepo.     

Effect of modified endogenous ethylene production on sex expression, bisexual flower development and fruit production in melon (Cucumis melo L.)

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.     

A Conserved Ethylene Biosynthesis Enzyme Leads to Andromonoecy in Two Cucumis Species

Andromonoecy is a widespread sexual system in angiosperms, characterized by plants carrying both male and bisexual flowers. Monoecy is characterized by the presence of both male and female flowers on the same plant. In cucumber, these sexual forms are controlled by the identity of the alleles at the M locus. In melon, we recently showed that the transition from monoecy to andromonoecy result from a mutation in 1-aminocyclopropane-1-carboxylic acid synthase (ACS) gene, CmACS-7. To isolate the andromonoecy gene in cucumber we used a candidate gene approach in combination with genetical and biochemical analysis. We demonstrated co-segregation of CsACS2, a close homolog of CmACS-7, with the M locus. Sequence analysis of CsACS2 in cucumber accessions identified four CsACS2 isoforms, three in andromonoecious and one in monoecious lines. To determine whether the andromonoecious phenotype is due to a loss of ACS enzymatic activity, we expressed the four isoforms in Escherichia coli and assayed their activity in vitro. Like in melon, 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 CsACS2 enzyme leads to the development of female flowers in monoecious lines, whereas a reduction of enzymatic activity yields hermaphrodite flowers. Consistent with this, CsACS2, like CmACS-7 in melon, is expressed specifically in carpel primordia of buds determined to develop carpels. Following ACS expression, inter-organ communication is likely responsible for the inhibition of stamina development. In both melon and cucumber, flower unisexuality seems to be the ancestral situation, as the majority of Cucumis species are monoecious. Thus, the ancestor gene of CmACS-7/CsACS2 likely have controlled the stamen development before speciation of Cucumis sativus (cucumber) and Cucumis melo (melon) that have diverged over 40 My ago. The isolation of the genes for andromonoecy in Cucumis species provides a molecular basis for understanding how sexual systems arise and are maintained within and between species.     

The Control of Sex Expression in Cucurbits by Ethephon

Application of ethephon to field-grown plants of both bush andtrailing forms of Cucurbita maxima and C. pepo caused leaf epinasty,suppression of male flowers and earlier production and increasein numbers of female flowers. This gave rise to an increasein the ratio of female to male flowers per plant and a decreasein the total number of flowers. Observations of C. pepo showed that even at the two true leafstage there are several nodes present in the unexpanded shoot.Each node has one main and several secondary buds. The sex ofthe main bud at the first five to six nodes is usually determinedat this stage but the secondary buds still have bisexual potential.The change in sex expression was brought about by all male flowerbuds that had formed by the spraying time aborting, and allbuds that developed (both main and secondary) for at least 7days after spraying became female flowers. Thus, nodes fiveand six had male flowers in the controls, whereas in ethephon-sprayedplants the presumptive male flowers aborted at the bud stageat these nodes and secondary primordia developed into functionalfemale flowers. Cucurbita maxima, Cucurbita pepo, sex expression, ethephon, ethylene, flower abortion, flower differentiation     

Involvement of Ethylene Biosynthesis and Signalling in the Transition from Male to Female Flowering in the Monoecious Cucurbita pepo

It is well established that ethylene is the main hormonal regulator of sexual expression in the Cucurbitaceae family, controlling not only the sexual fate of individual floral buds, but also the female flower transition, that is, the time at which the first female flower appears and therefore the number of female flowers per plant. Although sex determination of individual flower buds is known to be controlled by specific ethylene biosynthesis ACS genes in melon and cucumber, the role of ethylene genes in the control of the transition to female flowering is still unknown. We have identified two contrasting monoecious inbred lines of Cucurbita pepo, Bolognese (Bog) and Vegetable spaghetti (Veg), which differ in female flower transition but not in flower development. In Bog, which is very sensitive to ethylene, the transition to female flowering is very early, whereas in Veg, which is much less sensitive to ethylene, the transition occurs much later. In this article we compare the production of ethylene and the expression profiles of seven genes involved in the biosynthesis, perception, and signalling of ethylene in the two contrasting lines. Bog, with earlier female flower transition, showed higher ethylene production and CpACO1 expression in the apex at an earlier stage of plant development, when Bog is already producing female flowers, but Veg has not transitioned to female flowering yet. Moreover, the expression of the ethylene receptor and CTR-like genes in the apex of Veg and Bog plants indicates that these genes negatively regulate female flower transition during the earlier stages of plant development. The earlier transition to female flowering in Bog is not only associated with a higher production of ethylene in the apex but also with a premature decline of ethylene negative regulators (receptors and CTR-like) in the apex of the plant. These results provide the basis for a model that explains the regulation of female flowering transition in monoecious cucurbits.     

A positive feedback loop mediated by CsERF31 initiates female cucumber flower development

Sex determination is a crucially important developmental event that is pervasive throughout nature and enhances the adaptation of species. Among plants, cucumber (Cucumis sativus L.) can generate both unisexual and bisexual flowers, and the sex type is mainly controlled by several 1-aminocyclopropane-1-carboxylic acid synthases (CsACSs). However, the regulatory mechanism of these synthases remains elusive. Here, we used gene expression analysis, protein–DNA interaction assays, and transgenic plants to study the function of a gynoecium-specific gene, ETHYLENE RESPONSE FACTOR31 (CsERF31), in female flower differentiation. We found that in a predetermined female flower, ethylene signaling activates CsERF31 by CsEIN3, and then CsERF31 stimulates CsACS2, which triggers a positive feedback loop to ensure female rather than bisexual flower development. A similar interplay is functionally conserved in melon (Cucumis melo L.). Knockdown of CsERF31 by RNAi causes defective bisexual flowers to replace female flowers. Ectopic expression of CsERF31 suppresses stamen development and promotes pistil development in male flowers, demonstrating that CsERF31 functions as a sex switch. Taken together, our data confirm that CsERF31 represents the molecular link between female–male determination and female–bisexual determination, and provide mechanistic insight into how ethylene promotes female flowers, rather than bisexual flowers, in cucumber sex determination.

A key regulator promotes female flower development by triggering a positive feedback loop during cucumber sex determination.     

The role of ethylene and brassinosteroids in the control of sex expression and flower development in <Emphasis Type="Italic">Cucurbita pepo</Emphasis>

In this paper we compare the sensitivity of different squash genotypes to ethylene and brassinosteroids by studying the effects of different ethylene and brassinosteroid treatments on the sexual expression and flower development of different C. pepo genotypes: Bolognese (Bog) and Vegetable Spaghetti (Veg), two contrasting lines for ethylene production and sensitivity, as well as Cora, a standard commercial hybrid. Results have demonstrated that ethylene has a much greater effect on sexual expression and flower development in C. pepo than brassinosteroids. Ethephon increases the number of female flowers per plant and reduces the first male phase of development, while treatments with the ethylene inhibitors AVG and STS reduce the number of female flowers per plant and expand the first male phase of development. The differential response observed between genotypes appears to be related to ethylene production and sensitivity. Bog, which produces more ethylene and is more sensitive to this hormone, responded much better to AVG and STS, reducing the number of female flowers per plant, while Veg, which is characterised by lower production of and sensitivity to ethylene, responded better to ethephon by reducing the first male phase of development and increasing the number of female flowers per plant. The differential abortion of female and male flowers in ethephon, AVG and STS treatments, as well as the occurrence of bisexual flowers in the AVG and STS treated plants of the more ethylene sensitive genotypes, demonstrate that ethylene is also involved in the development of female flowers. Female flower buds require a minimal level of ethylene not only to complete their development and maturation without a premature abortion, but also to arrest the development of stamens in the third whorl and to promote the appropriate growth of the carpels. On the contrary, the role of brassinosteroids in the sexual expression of C. pepo was not so evident. The application of brassinazole, an inhibitor of brassinosteroid biosynthesis slightly changes the production of ethylene in the three analysed genotypes, but those changes have little effect on their sexual phenotypes, and they do not alter the development of the unisexual flowers.     

Correlation between development of female flower buds and expression of the CS-ACS2 gene in cucumber plants

Ethylene plays a key role in sex determination of cucumber flowers. Gynoecious cucumber shoots produce more ethylene than monoecious shoots. Because monoecious cucumbers produce both male and female flower buds in the shoot apex and because the relative proportions of male and female flowers vary due to growing conditions, the question arises as to whether the regulation of ethylene biosynthesis in each flower bud determines the sex of the flower. Therefore, the expression of a 1-aminocyclopropane-1-carboxylic acid synthase gene, CS-ACS2, was examined in cucumber flower buds at different stages of development. The results revealed that CS-ACS2 mRNA began to accumulate just beneath the pistil primordia of flower buds at the bisexual stage, but was not detected prior to the formation of the pistil primordia. In buds determined to develop as female flowers, CS-ACS2 mRNA continued to accumulate in the central region of the developing ovary where ovules and placenta form. In gynoecious cucumber plants that produce only female flowers, accumulation of CS-ACS2 mRNA was detected in all flower buds at the bisexual stage and at later developmental stages. In monoecious cucumber, flower buds situated on some nodes accumulated CS-ACS2 mRNA, but others did not. The proportion of male and female flowers in monoecious cucumbers varied depending on the growth conditions, but was correlated with changes in accumulation of CS-ACS2 mRNA in flower buds. These results demonstrate that CS-ACS2-mediated biosynthesis of ethylene in individual flower buds is associated with the differentiation and development of female flowers.     

乙烯利诱导网纹甜瓜主蔓两性花花芽分化的形态和解剖学研究

以网纹甜瓜(Cucumis melo L.var. reticulatus Naud.)品种’西域1号’为试验材料,于幼苗3叶1心期喷施浓度为150 mg·L^-1的乙烯利溶液进行处理,诱导主蔓形成两性花,以清水为对照,分别对处理和对照植株不同时期的主蔓和侧蔓花芽分化过程进行形态和解剖学观察。结果表明：经乙烯利处理后,幼苗植株主蔓花原基持续向两性花分化,最终发育形成两性花。未经处理植株的主蔓花原基在分化早期与两性花发育过程相同,但在雌蕊出现后,不再继续发育,最终发育形成雄花。处理植株主蔓两性花发育过程与侧蔓两性花发育过程相同。     

Characterization of ethylene effects on sex determination in cucumber plants

Sex differentiation in cucumber plants (Cucumis sativus L.) appears to be determined by the selective arrest of the stamen or pistil primordia. We investigated the influence of an ethylene-releasing agent (ethephon) or an inhibitor of ethylene biosynthesis (aminoethoxyvinyl glycine) on sex differentiation in different developmental stages of flower buds. These treatments influence sex determination only at the stamen primordia differentiation stage in both monoecious and gynoecious cucumbers. To clarify the relationships between the ethylene-producing tissues and the ethylene-perceiving tissues in inducing female flowers in the cucumber, we examined the localization of mRNA accumulation of both the ACC synthase gene (CS-ACS2) and the ethylene-receptor-related genes (CS-ETR1, CS-ETR2, and CS-ERS) in flower buds by in situ hybridization analysis. CS-ACS2 mRNA was detected in the pistil primordia of gynoecious cucumbers, whereas it was located in the tissues just below the pistil primordia and at the adaxial side of the petals in monoecious cucumbers. In flower buds of andromonoecious cucumbers, only CS-ETR1 mRNA was detected, and was located in the pistil primordia. The localization of the mRNAs of the three ethylene-receptor-related genes in the flower buds of monoecious and gynoecious cucumbers overlap but are not identical. We discuss the relationship between the mRNA accumulation patterns and sex expression in cucumber plants.     

Floral primordia-targeted ACS (1-aminocyclopropane-1-carboxylate synthase) expression in transgenic Cucumis melo implicates fine tuning of ethylene production mediating unisexual flower development

Main conclusion

Floral primordia-targeted expression of the ethylene biosynthetic gene, ACS , in melon suggests that differential timing and ethylene response thresholds combine to promote carpels, inhibit stamens, and prevent asexual bud formation. Typical angiosperm flowers produce both male and female reproductive organs. However, numerous species have evolved unisexuality. Melons (Cucumis melo L.) can produce varying combinations of male, female or bisexual flowers. Regardless of final sex, floral development begins with sequential initiation of all four floral whorls; unisexuality results from carpel or stamen primordia arrest regulated by the G and A loci, respectively. Ethylene, which promotes femaleness, is a key factor regulating sex expression. We sought to further understand the location, timing, level, and relationship to sex gene expression required for ethylene to promote carpel development or inhibit stamen development. Andromonoecious melons (GGaa) were transformed with the ethylene biosynthetic enzyme gene, ACS (1-aminocyclopropane-1-carboxylate synthase), targeted for expression in stamen and petal, or carpel and nectary, primordia using Arabidopsis APETALA3 (AP3) or CRABSCLAW (CRC) promoters, respectively. CRC::ACS plants did not exhibit altered sex phenotype. AP3::ACS melons showed increased femaleness manifested by gain of a bisexual-only phase not seen in wild type, decreased male buds and flowers, and loss of the initial male-only phase. In extreme cases, plants became phenotypically hermaphrodite, rather than andromonoecious. A reduced portion of buds progressed beyond initial whorl formation. Both the ACS transgene and exogenous ethylene reduced the expression of the native carpel-suppressing gene, G, while elevating expression of the stamen-suppressing gene, A. These results show ethylene-mediated regulation of key sex expression genes and suggest a mechanism by which temporally regulated ethylene production and differential ethylene response thresholds can promote carpels, inhibit stamens, and prevent the formation of asexual buds.     

Molecular characterization and isolation of the <Emphasis Type="Italic">F/f</Emphasis> gene for femaleness in cucumber (<Emphasis Type="Italic">Cucumis sativus</Emphasis> L.)

The biological processes leading to sex expression in plants are of tremendous practical significance for fruit production of many agricultural and horticultural crops. Sex-expression studies in cucumber showed that the different sex types are determined by three major genes: M/m, F/f and A/a. The M/m gene in the dominant condition suppresses stamina development and thus leads to female flowers. The F/f gene in the dominant condition shifts the monoecious sex pattern downwards and promotes femaleness by causing a higher level of ethylene in the plant. To investigate the molecular character of the gene F/f, we used nearly isogenic gynoecious (MMFF) and monoecious (MMff) lines (NIL) produced by our own backcross programme. Our investigations confirmed the result of other groups that an additional genomic ACC synthase (key enzyme of ethylene biosynthesis) sequence (CsACS1G) should exist in gynoecious genotypes. A linkage was also verified between the F/f locus and the CsACS1G sequence with our plant material. After the exploration of different Southern hybridization patterns originating from different CsACS1 probes, a restriction map of the CsACS1 locus was constructed. By using this restriction map, the duplication of the CsACS1 gene and following mutation of the CsACS1G gene could be explained. The promoter regions of the genes CsACS1G and CsACS1 were amplified in a splinkerette PCR and sequenced. An exclusive amplification of the new isolated sequence (CsACS1G) in gynoecious (MMFF) and sub-gynoecious (MMFf) genotypes confirmed that the isolated gene is the dominant F allele.     

Sex expression in floral buds of Acorn squash

Summary Staminate flowers of the Acorn squash (Cucurbita pepo, cv. Table Queen) are initiated monosexually, pistillate flowers are initiated bisexually. Pistillate flower primordia are physiologically bisexual from a stage below 0.3 mm until they are 0.8 mm in width.Male and female floral buds were cultivated on White's medium, modified according to Galun et al. (1963). In excised female floral buds the relative development of stamens is better than in in-situ floral buds; the opposite relationship holds for ovary development. The smaller the bud at excision, the greater the stamen growth and the lesser the ovary growth, both in absolute and relative terms. Although smaller pistillate floral buds cultured in vitro showed a greater increase in maleness than larger ones, there was no difference in the percentage of female buds between explants of the two sizes when originating from the mixed stem region (that is, the stem region carrying male and female flowers). Addition of 3-indoleacetic acid and the anti-auxin p-chlorophenoxyisobutyric acid to standard medium in equal amounts resulted in somewhat better development of excised floral buds than addition of each of these compounds alone.The percentage of female buds among explants originating from identical nodes was not significantly influenced by 3-indoleacetic acid or p-chlorophenoxyisobutyric acid.Weizmann Memorial Fellow. The studies reported in this publication were mainly undertaken at the Weizmann Institute of Science     

A Major Gene Conferring Reduced Ethylene Sensitivity and Maleness in Cucurbita pepo

External treatment with ethylene had indicated earlier that this hormone is the main factor controlling sex determination in Cucurbita pepo. Up to now, however, there was no genetic evidence that supported the relationship between ethylene production, or perception, and sexual expression in this species. Here we demonstrate that the extreme male phenotype of the Vegetable Spaghetti (Veg) inbred line of C. pepo subspecies pepo is determined by a major gene that confers reduced ethylene sensitivity in plants. The production of female flowers in the Veg line is very delayed and reduced with respect to the contrasting Bolognese (Bog) line, ranging between 5 and 35% of female flowers per plant. This enhanced maleness trait segregates as a single gene in the F₂ and backcross (BC) generations, and co-segregates with a weak ethylene-insensitive phenotype in the F₂ population, suggesting that the gene responsible for the Veg phenotype could be the result of a mutation in a receptor or response gene for ethylene. Although the etiolated seedlings of the Veg line, and the most androecious plants in the F₂ generation, produce more ethylene than those of the contrasting line, they are less sensitive to this hormone, as indicated by a weaker triple response and a delayed abscission of ethylene-treated male flowers. Given that the sexual phenotype of F₂ plants is correlated with ethylene sensitivity, with the more sensitive plants producing the higher number of female flowers, our results demonstrate that the ethylene response is directly involved in the control of sex determination in C. pepo. It regulates the induction of female flower production, and therefore the extension of the initial phase of development in which the plant produces only male flowers, as well as the number of female flowers per plant.     

C(2)H(4) metabolism in morning glory flowers

Flowers of Ipomoea tricolor Cav. (cv. Heavenly Blue) were cut at various stages of development and evaluated for their ability to metabolize ethylene. Freshly cut buds or flowers were treated in glass containers for 8 hours with 6 μl/liter of highly purified ¹⁴C₂H₄. Following removal of dissolved ¹⁴C₂H₄, radioactivity was determined for the different flower tissues and trappd CO₂. ¹⁴C₂H₄ oxidation to ¹⁴CO₂ and tissue incorporation occurred at very low to nondetectable levels 2 to 3 days prior to flower opening. About 1 day prior to full bloom, just at the time when mature buds become responsive to ethylene (Kende and Hanson, Plant Physiol 1976, 57: 523-527), there was a dramatic increase in the capacity of the buds to oxidize ¹⁴C₂H₄ to ¹⁴CO₂. This activity continued to increase until the flower was fully opened reaching a peak activity of 2,500 dpm per three flowers per 8 hours. It then declined as the flower closed and rapidly senesced. A similar but smaller peak occurred in tissue incorporation and it was followed by a second peak during late flower senescence. This first peak in tissue incorporation and the dramatic peak in ethylene oxidation slightly preceded a large peak of natural ethylene production which accompanied flower senescence. The ethylene metabolism observed was clearly dependent on cellular metabolism and did not involve microorganisms since heat killing destroyed this activity and badly contaminated heat-killed flowers were unable to metabolize ethylene.     

Epigenetic control of sexual phenotype in a dioecious plant,Melandrium album

Melandrium album (syn.Silene latifolia) is a model dioecious species in which theY chromosome, present only in heterogametic males, plays both a male-determining and a strict female-suppressing role. We showed that treatment with 5-azacytidine (5-azaC) induces a sex change to androhermaphroditism (andromonoecy) in about 21% of male plants, while no apparent phenotypic effect was observed in females. All of these bisexual androhermaphrodites (with the standard male 24,AA +XY karyotype) were mosaics possessing both male and hermaphrodite flowers and, moreover, the hermaphrodite flowers displayed various degrees of gynoecium development and seed setting. Southern hybridization analysis with a repetitive DNA probe showed that the 5-azacytidine-treated plants were significantly hypomethylated in CG doublets, but only to a minor degree in CNG triplets. The bisexual trait was transmitted to two successive generations, but only when androhermaphrodite plants were used as pollen donors. The sex reversal was inherited with incomplete penetrance and varying expressivity. Based on the uniparental inheritance pattern of androhermaphroditism we conclude that it originated either by 5-azaC induced inhibition ofY-linked female-suppressing genes or by a heritable activation of autosomal female-determining/promoting genes which can be reversed, on passage through female meiosis, by a genomic imprinting mechanism. The data presented indicate that female sex suppression inM. album XY males is dependent on methylation of specific DNA sequences and can be heritably modified by hypomethylating drugs.     

The influence of ethylene perception on sex expression in melon (<Emphasis Type="Italic">Cucumis melo </Emphasis>L.) as assessed by expression of the mutant ethylene receptor, <Emphasis Type="Italic">At-etr1-1</Emphasis>, under the control of constitutive and floral targeted promoters

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.     

Adventitious staminate flower formation in gibberellin treated gynoecious cucumber plants

Single gibberellin (A₄₊₇) treatments induced the appearanceof staminate floral buds in several consecutive nodes on themain stem of genetically female cucumber (Cucumis sativus L.).The staminate buds appeared next to pistillate buds which showedvarious degrees of degeneration. Similarly, repeated GA treatmentsinduced the appearance of staminate flowers in otherwise strictlyhermaphrodite plants, next to bisexual flowers. However, thebisexual buds, unlike the pistillate ones, did not show anydeleterious effects of the GA treatment. Therefore, it is inferredthat the hormonally induced staminate buds did not develop bysexual reversion of would-be pistillate or bisexual buds, butrather, represent adventitious buds which, in normally grownfemale or hermaphrodite plants, never develop. It thus seemsthat predetermined pistillate or bisexual buds do not changeinto staminate ones, while change in the reverse direction hasbeen demonstrated in the past (at least for the gynoecious ones). The effectiveness of the GA treatment in the gynoecious plantsshowed an acropetal gradient both within the affected region,as well as along the main stem. Autoradiographic histologicalexaminations showed that the course of development of the inducedstaminate floral bud did not differ from that of normally developingbuds. (Received June 16, 1977; )