Expression of ethylene biosynthetic and receptor genes in rose floral tissues during ethylene-enhanced flower opening

Ethylene production, as well as the expression of ethylene biosynthetic (Rh-ACS1-4 and Rh-ACO1) and receptor (Rh-ETR1-5) genes, was determined in five different floral tissues (sepals, petals, stamens, gynoecia, and receptacles) of cut rose (Rosa hybrida cv. Samantha upon treatment with ethylene or the ethylene inhibitor 1-methylcyclopropene (1-MCP). Ethylene-enhanced ethylene production occurred only in gynoecia, petals, and receptacles, with gynoecia showing the greatest enhancement in the early stage of ethylene treatment. However, 1-MCP did not suppress ethylene production in these three tissues. In sepals, ethylene production was highly decreased by ethylene treatment, and increased dramatically by 1-MCP. Ethylene production in stamens remained unchanged after ethylene or 1-MCP treatment. Induction of certain ethylene biosynthetic genes by ethylene in different floral tissues was positively correlated with the ethylene production, and this induction was also not suppressed by 1-MCP. The expression of Rh-ACS2 and Rh-ACS3 was quickly induced by ethylene in gynoecia, but neither Rh-ACS1 nor Rh-ACS4 was induced by ethylene in any of the five tissues. In addition, Rh-ACO1 was induced by ethylene in all floral tissues except sepals. The induced expression of ethylene receptor genes by ethylene was much faster in gynoecia than in petals, and the expression of Rh-ETR3 was strongly suppressed by 1-MCP in all floral tissues. These results indicate that ethylene biosynthesis in gynoecia is regulated developmentally, rather than autocatalytically. The response of rose flowers to ethylene occurs initially in gynoecia, and ethylene may regulate flower opening mainly through the Rh-ETR3 gene in gynoecia.     

Role of the gynoecium in natural senescence of carnation (Dianthus caryophyllus L.) flowers

Although the role of the gynoecium in natural senescence of the carnation flower has long been suggested, it has remained a matter of dispute because petal senescence in the cut carnation flower was not delayed by the removal of gynoecium. In this study, the gynoecium was snapped off by hand, in contrast to previous investigations where removal was achieved by forceps or scissors. The removal of the gynoecium by hand prevented the onset of ethylene production and prolonged the vase life of the flower, demonstrating a decisive role of the gynoecium in controlling natural senescence of the carnation flower. Abscisic acid (ABA) and indole-3-acetic acid (IAA), which induced ethylene production and accelerated petal senescence in carnation flowers, did not stimulate ethylene production in the flowers with gynoecia removed (-Gyn flowers). Application of 1-aminocyclopropane-1-carboxylate (ACC), the ethylene precursor, induced substantial ethylene production and petal wilting in the flowers with gynoecia left intact, but was less effective at stimulating ethylene production in the -Gyn flowers and negligible petal in-rolling was observed. Exogenous ethylene induced autocatalytic production of the gas and petal wilting in the -Gyn flowers. These results indicated that ethylene generated in the gynoecium triggers the onset of ethylene production in the petals of carnation during natural senescence.     

Ethylene perception is involved in female cucumber flower development

It is well established that ethylene promotes female flower development in cucumber. However, little is known about how the gaseous hormone selectively affects female flowers, and what mechanism it uses. Previously, we found organ‐specific DNA damage in the primordial anther of female cucumber flowers. This finding led to a hypothesis that ethylene might promote female flower development via the organ‐specific induction of DNA damage in primordial anthers. In this study, we tested this hypothesis first by demonstrating ethylene induction of DNA damage via the ethylene signaling pathway using cucumber protoplasts. Then, using representative component genes of the ethylene signaling pathway as probes, we found that one of the ethylene receptors, CsETR1, was temporally and spatially downregulated in the stamens of stage‐6 female cucumber flowers, especially along with the increase of the nodes. Furthermore, by constructing transgenic Arabidopsis plants with organ‐specific expression of antisense CsETR1 under the control of an AP3 promoter to downregulate ETR1 expression in the stamens, we generated Arabidopsis ‘female flowers’, in which the abnormal stamens mimic those of female cucumber flowers. Our data suggest that ethylene perception is involved in the arrest of stamen development in female cucumber flowers through the induction of DNA damage. This opens up a novel perspective and approach to solve the half‐century‐long puzzle of how gaseous ethylene selectively promotes female flowers in the monoecious cucumber plant.     

Characteristics of the inhibitory action of 1,1-dimethyl-4-(phenylsulfonyl)semicarbazide (DPSS) on ethylene production in carnation (Dianthus caryophyllus L.) flowers

1,1-Dimethyl-4-(phenylsulfonyl)semicarbazide (DPSS)inhibited ethylene productionin carnation flowers during natural senescence, butdid not inhibit the ethyleneproduction induced by exogenous ethylene in carnationflowers, by indole-3-acetic acid (IAA) in mungbean hypocotylsegments and by wounding in winter squashmesocarp tissue. These findings suggested that DPSSdoes not directly inhibit ethylene biosynthesis fromL-methionine to ethylenevia S-adenosyl-L-methionine and1-aminocyclopropane-1-carboxylate. During naturalsenescence of carnation flowers, abscisic acid (ABA)was accumulated in the pistil and petals 2 days beforethe onset of ethylene production in the flower, andthe ABA content remained elevated until the onset ofethylene production. Application of exogenousABA to cut flowers from the cut stem end caused arapid increase in the ABA content in flower tissuesand promoted ethylene production in the flowers. These results were in agreement with the previousproposal that ABA plays a crucial role in theinduction of ethylene production during natural senescence incarnation flowers. DPSS preventedthe accumulation of ABA in both the pistil and petals,suggesting that DPSS exerted its inhibitory action onethylene production in naturally-senescing carnationflowers through the effect on the ABA-related process.     

Chasmogamous and Cleistogamous Pollination in Salpiglossis sinuata

Pollen from chasmogamous flowers of Salpiglossis sinuata L. could not be induced to germinate in vitro unless stigmatal extract was applied to the culture medium. The substance that induces pollen germination in the stigmatal extract is water-soluble and heat-stable. Crosses could not be achieved between chasmogamous and cleistogamous flowers because of structural incompatibility. Pollinated pistils of chasmogamous flowers release a large amount of ethylene. The burst of ethylene release is due to an interaction between pollen tubes and stylar tissue and is directly proportional to the quantity of pollen placed on the stigma. Cleistogamous flower buds also produce a burst of ethylene at the time of pollination within the closed flower. The ethylene release may be a cause of reduced corolla development associated with cleistogamous flowers.     

Graft‐accelerated virus‐induced gene silencing facilitates functional genomics in rose flowers

Rose has emerged as a model ornamental plant for studies of flower development, senescence, and morphology, as well as the metabolism of floral fragrances and colors.Virus-induced gene silencing(VIGS) has long been used in functional genomics studies of rose by vacuum infiltration of cuttings or seedlings with an Agrobacterium suspension carrying TRV-derived vectors. However, VIGS in rose flowers remains a challenge because of its low efficiency and long time to establish silencing. Here we present a novel and rapid VIGS method that can be used to analyze gene function in rose,called ‘graft-accelerated VIGS', where axil ary sprouts are cut from the rose plant and vacuum infiltrated with Agrobacterium. The inoculated scions are then grafted back onto the plants to flower and silencing phenotypes can be observed within 5 weeks, post-infiltration. Using this new method, we successfully silenced expression of the RhDFR_1, RhA G, and RhNUDX_1 in rose flowers, and affected their color, petal number, as well as fragrance, respectively. This grafting method will facilitate high-throughput functional analysis of genes in rose flowers. Importantly, it may also be applied to other woody species that are not currently amenable to VIGS by conventional leaf or plantlet/seedling infiltration methods.     

Light and ethylene effects on assimilate distribution,acid invertase activity and keeping quality of begonia

Flowering plants of Begonia × cheimantha cv Emma and Begonia x hiemalis cv Schwabenland Red were exposed to different light levels (0, 40, 80 M m^–2S^–1) and to ethylene (150 nl 1^–1) in growth cabinets. Increasing irradiance level increased the number of flower buds in both begonia species. The amount of ¹⁴C-assimilates translocated to flower buds and the acid invertase activity in flower buds and flowers also increased with increasing irradiance level. Conversely, treatment with ethylene decreased the accumulation of ¹⁴C in flowers and flower buds, but did not affect acid invertase activity. Ethylene accelerated abscission of flowers and flower buds and increased the number of cup shaped and small flowers.     

A simple,economical, and high efficient protocol to produce in vitro miniature rose

Due to its high commercial value, many studies on rose (Rosa hybrida L.) micropropagation have been published. However, there are a limited number of studies on rose in vitro flowering. These studies only focused on the roles of plant growth regulators in the formation and morphogenesis of flowers. In this protocol, cytokinin was confirmed to positively function in the induction of in vitro rose flowers. In fact, more than 40% of in vitro shoots were induced to flower when they were grown on a medium supplemented with benzylaminopurine (BA) (2 mg L⁻¹) and IAA (0.1 mg L⁻¹). In addition, this study showed that the growth medium supplemented with only coconut water (15 or 20% v/v) was very efficient to induce flowering of in vitro miniature rose plants (> 70%) after 60 d of subculture. In addition, the in vitro flowers were normal and almost similar to ex vitro flowers in terms of flower shape and color. Based on these results, a detailed procedure for in vitro miniature rose flower production is provided.

     

Role of Aminocyclopropane-1-carboxylic Acid (ACC) in Control of Ethylene Production in Fresh and Cold-stored Rose Flowers

The relationships between ethylene production, aminocyclopropane-1-carboxylicacid (ACC) content and ethylene-forming-enzyme (EFE) activityduring ageing and cold storage of rose flower petals (Rose hybridaL. cv. Gabriella) were investigated. During flower ageing at20 °C there was a climacteric rise in petal ethylene production,a parallel increase in ACC content, but a continuous decreasein EFE activity. Applied ACC increased petal ethylene productionc. 200-fold. During cold storage of flowers at 1 °C therewere parallel increases in petal ethylene production and ACCcontent, to levels greater than those reached in fresh flowersheld at 20 °C. EFE activity decreased during storage. Immediatelyafter cold-stored flowers were transferred to 20 °C ethyleneproduction and ACC levels were c. four times greater than infreshly cut flowers. These levels increased to maximum valuesof two to four times the maximum values reached during ageingof fresh, unstored, flowers. It was concluded that in rose petalsethylene synthesis is probably regulated by ACC levels and thatcold storage stimulates ethylene synthesis because it increasesthe levels of ACC in the petals. Key words: Rose flower, senescence, ethylene     

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.     

Role of 1-aminocyclopropane-1-carboxylic acid in the control of female flowering in Cucurbita pepo

Cucurbita pepo L. cv. Trailing Marrow is monoecious, bearing separate male and female flowers and the first functional flowers are usually male. Treatment with 300 ppm ethephon delayed and greatly reduced male flower production and also increased female flower numbers. When plants were sprayed with aminoethoxyvinylglycine (AVG) no female flowers were produced but male flower production was unaffected. Even when ethephon was applied to AVG-treated plants there was still complete inhibition of female flower production. Similarly, AVG-treated plants subsequently exposed to 4000 ppm ethylene for two days never produced female flowers. AVG inhibits the penultimate stage in ethylene biosynthesis i.e. immediately before 1-aminocyclopropane-1-carboxylic acid (ACC). Although spraying AVG-treated plants with ACC did not reverse the inhibition, application of ACC via a cut petiole for a 72 h period following AVG application did cause female flowers to form. The evidence indicates that ACC and not ethylene is the factor controlling female flower production in C. pepo .     

Inhibition of wilting and autocatalytic ethylene production in cut carnation flowers by cis-propenylphosphonic acid

The effect of cis-propenylphosphonic acid (PPOH), a structural analoge of ethylene, on flower wilting and ethylene production was investigated using cut carnation flowers which are very sensitive to ethylene. Wilting (petal in-rolling) of the flowers was delayed by continuously immersing the stems in a 5–20 mM PPOH solution. In addition, the continuous treatment with PPOH markedly reduced autocatalytic ethylene production of the petals accompanying senescence. This reduction of autocatalytic ethylene production was considered responsible for the inhibitory effect of PPOH on flower wilting. The inhibitory activity of trans-propenylphosphonic acid (trans-PPOH), on both flower wilting and the autocatalytic ethylene production accompanying senescence was markedly lower than that of PPOH, suggesting that PPOH action is stereoselective. PPOH may be of interest as a new, water-soluble inhibitor of wilting and autocatalytic ethylene production in cut carnation flowers.