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.     

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 production and β-cyanoalanine synthase activity in carnation flowers

The relationship between ethylene production and the CN^--assimilating enzyme -cyanoalanine synthase (CAS; EC 4.4.1.9) was examined in the carnation (Dianthus caryophyllus L.) flower. In petals from cut flowers aged naturally or treated with ethylene to accelerate senescence the several hundred-fold increase in ethylene production which occurred during irreversible wilting was accompanied by a one- to twofold increase in CAS activity. The basal parts of the petal, which produced the most ethylene, had the highest CAS activity. Studies of flower parts (styles, ovaries, receptacles, petals) showed that the styles had a high level of CAS together with the ethylene-forming enzyme (EFE) system for converting 1-aminocyclopropane-1-carboxylic acid (ACC) to ethylene. The close association between CAS and EFE found in styles could also be observed in detached petals after induction by ACC or ethylene. Treatment of the cut flowers with cycloheximide reduced synthesis of CAS and EFE. The data indicate that CAS and ethylene production are associated, and are discussed in relation to the hypothesis that CN^- is formed during the conversion of ACC to ethylene.Abbreviations ACC 1-aminocyclopropane-1-carboxylic acid - AVG aminoethoxyvinylglyoine - CAS -cyanoalanine synthase - CHI cycloheximide - EFE ethylene-forming enzyme     

Lack of Control by Early Pistillate Ethylene of the Accelerated Wilting of Petunia hybrida Flowers

Well before pollen tube penetration, ethylene has begun to disseminate from pollinated styles of Petunia hybrida flowers. Previous stigmatic application of aminoethoxyvinylglycine (AVG) completely prevented this ethylene synthesis, indicating that the endogenous 1-aminocyclopropane-1-carboxylic acid (ACC) in pollen is not readily converted on the stigma. Compared to other flower parts, the capacity of the ethylene forming enzyme was largest in the stigma. When applied to the stigma, ACC caused ethylene synthesis, but did not accelerate wilting, unless high concentrations (20 nanomols) were used. Upon pollination or stigma wounding, the early ethylene evolved exclusively from the gynoecium, much later followed by the synthesis of corolla ethylene. Employing wideneck Erlenmeyer flasks, the competitive inhibitor of ethylene action, norbornadiene, was applied to entire flowers in situ, with delaying effects on wound-induced wilting. In contrast, norbornadiene treatment of styles alone, using capillaries, could not postpone wilting. Pollination with foreign pollen species did not lead to accelerated corolla wilting, notwithstanding considerable synthesis of ethylene during the first 5 hours. In situ treatment of the stigma with AVG considerably delayed wound- and pollination-induced wilting. Removal of the entire AVG-treated style 6 hours after stigma wounding still allowed for the postponement of the accelerated wilting, even at very low concentrations of AVG. It is concluded that early stylar ethylene does not play a role in the acceleration of wilting but that, much later, corolla ethylene does, induced by a mobile wilting factor from the stigma, which is ACC.     

Role of ethylene in the senescence of isolated hibiscus petals

Senescence of petals isolated from flowers of Hibiscus rosa-sinensis L. (cv Pink Versicolor) was associated with increased ethylene production. Exposure to ethylene (10 microliters per liter) accelerated the onset of senescence, as indicated by petal in-rolling, and stimulated ethylene production. Senescence was also hastened by basal application of 1-aminocyclopropane-1-carboxylic acid (ACC). Aminooxyacetic acid, an inhibitor of ethylene biosynthesis, effectively inhibited ethylene production by petals and delayed petal in-rolling. In marked contrast to these results with mature petals, immature petals isolated from flowers the day before flower opening did not respond to ethylene in terms of an increase in ethylene production or petal in-rolling. Furthermore, treatment with silver thiosulfate the day before flower opening effectively prevented petal senescence, while silver thiosulfate treatment on the morning of flower opening was ineffective. Application of ACC to both immature and mature petals greatly stimulated ethylene production indicating the presence of an active ethylene-forming enzyme in both tissues. Immature petals contained less free ACC than mature, presenescent petals and appeared to possess a more active system for converting ACC into its conjugated form. Thus, while the nature of the lack of responsiveness of immature petals to ethylene is unknown, ethylene production in hibiscus petals appears to be regulated by the control over ACC availability.     

Programme of senescence in petals and carpels of Pisum sativum L. flowers and its control by ethylene

The role of ethylene in the control of senescence of both petals and unpollinated carpels of pea was investigated. An increase in ethylene production accompanied senescence, and the inhibitors of ethylene action were effective in delaying senescence symptoms in different flower verticils. Pollination did not seem to trigger the senescence syndrome in the corolla as deduced from the observation that petals from pollinated and unpollinated flowers and from flowers whose carpels had been removed senesced at the same time. A cDNA clone encoding a putative ethylene-response sensor (psERS) was isolated from pea flowers, and the pattern of expression of its mRNA was studied during development and senescence of different flower tissues. The levels of psERS mRNA paralleled ethylene production (and also levels of 1-aminocyclopropane-1-carboxylic acid oxidase (ACO) mRNA) in both petals and styles. Silver thiosulfate treatments were efficient at preventing ACO and psERS mRNA induction in petals. However, the same inhibitor showed no ability to modify expression patterns in pea carpels around the anthesis stage, suggesting different controls for ethylene synthesis and sensitivity in different flower organs. Received: 18 June 1998 / Accepted: 22 December 1998     

Pollination-induced senescence of Phalaenopsis petals

The longevity of cut Phalaenopsis (Phalaenopsis hybrid, cv. Herbet Hager) flowers is normally 2 to 3 weeks. After pollination however, there was a rapid acceleration of the wilting process, beginning after only 24 h. Enhancement of senescence in several Phalaenopsis cultivars as well as in Doritaenopsis, Dendrobium and Cymbidium, was induced by successful pollination and only slightly or not at all by emasculation. Wilting of the flowers was accompanied by a loss of water from cells of the upper layer of the petals, leading to their upward folding. Following pollination there was an increase in ethylene production and sensitivity to ethylene. The increase in ethylene production began about 10 h after pollination and reached its peak after 30 h. An obvious increase in sensitivity to ethylene could already be detected 4 h after pollination and reached its peak 10 h after pollination. The increase en ethylene sensitivity following pollination was not dependent on endogenous ethylene production as it occurred also in flowers treated with (aminooxy)acetic acid, an inhibitor of ethylene biosynthesis.Abbreviations AOA = (aminooxy)acetic acid - RH = relative humidity - SEM = scanning electron microscope     

Effects of pollination on floral attraction and longevity

The end of a flower's attraction to pollinators may be due toa range of visible cues such as permanent flower closure, acolour change, and withering or abscission of the petals. Floralattraction may be reduced by pollination. Pollination-inducedconclusion of floral attraction is often due to a colour changeor to flower closure. This may or may not be followed by a reductionin floral longevity, defined as the time to petal withering,wilting or shattering. In a few species floral longevity isincreased following pollination-induced flower closure or apollination-induced change in colour. Floral attraction, therefore,has to be disting uished from floral longevity. A literature survey shows that pollination rapidly reduces floralattraction in numerous orchids, but among other plant familiesonly about 60 genera have been found to show pollination-inducedshortening of floral attraction. Although only a few specieshave been investigated, it was invariably established that theeffect of pollination is blocked by inhibitors of ethylene synthesisor ethylene perception, hence is mediated by ethylene. The flowersthat cease to be attractive to pollinators, shortly followingpollination, tend to be from families that are known mainlyto comprise species in which flower longevity, petal colour,or flower closure, is sensitive to exogenous ethylene. Thisindicates that the effect of pollination on floral attractionis generally mediated by endogenous ethylene. Numerous species reportedly show a decrease in the period offloral attraction after exposure to ethylene, whereas only fora small number of species a decrease in the period of floralattraction induced by pollination has been observed. This discrepancymay be due to the greater attention that has been paid to theeffects of ethylene. Nonetheless, the possibility remains thatendogenous ethylene has a role in changing perianth form andcolour in addition to signalling the occurrence of pollination. Key words: Ethylene sensitivity, flower closure, flower longevity, pollination, petal colour, petal wilting, petal withering, petal abscission     

Changes in ethylene production and 1-aminocyclopropane-1-carboxylic acid content of pollinated carnation flowers

Pollination of flowers of standard carnation (Dianthus caryophyllus L. cv. White Sim) with pollen from flowers of miniature carnations (D. caryophyllus L. cv. Exquisite) caused them to wilt irreversibly within 1 to 2 days. Pollination stimulated a sequential increase in ethylene production by stigmas, ovaries, receptacles, and petals of the flowers. The ACC content of the stigmas increased rapidly in the first few hours after pollination. The possibility that subsequent production of ethylene by other parts of the flower is stimulated by translocated ACC is discussed. Ethylene production and ACC content of other parts of the flower reached their maximum 24 h after pollination. The petal tissues contributed the bulk of the ethylene productionper flower thereafter. There appears to be a qualitative difference between the enzyme in the stigmas converting ACC to ethylene and that in other parts of the flower.     

Role of short-chain saturated fatty acids in the control of ethylene sensitivity in senescing carnation flowers

In cut carnations ( Dianthus caryophyllus L. cv. Cally). petal senescence was associated with a climacteric pattern in ethylene production and an increase in ethylene sensitivity during the preclimacteric stage. The increase in ethylene sensitivity was caused by short-chain saturated fatty acids (C₇ to C₁₀) produced in the petals during the early stages of senescence. Pollination or application of octanoic acid to the styles of unpollinated flowers resulted in a sudden increase in ethylene sensitivity and a marked acceleration of senescence. Treatment with silver thiosulfate (STS) resulted in a suppression of ethylene sensitivity and a marked reduction in the levels of these fatty acids. However, even in STS-treated flowers pollination or treatment with octanoic acid gave rise to a drastic increase in ethylene sensitivity. Exposure of carnation flowers to 2. 5-norbornadicne (NBD) vapours resulted in a dramatic suppression of ethylene sensitivity which was also overridden by stylar application of octanoic acid. Exposure to NBD suppressed the increase in ethylene sensitivity caused by treatment with octanoic acid. It appears that short-chain saturated fatty acids increased ethylene sensitivity by increasing the ability of the tissue to bind ethylene.     

Does Pollination Induce Corolla Abscission of Cyclamen Flowers by Promoting Ethylene Production?

Very low ethylene production rates were measured in nonpollinated Cyclamen persicum Mill flowers, and no change in production was observed during the whole life span of the flower until death. Normal senescence was accompanied by a gradual discoloration and loss of turgor followed by wilting. Pollination induced a dramatic increase in ethylene evolution, culminating in a peak 4 days after pollination, and abscission of the corolla on that day. Silver-thiosulfate, an inhibitor of ethylene action, had no effect on longevity of unpollinated flowers, but completely nullified the effect of pollination on corolla abscission. Exposing unpollinated flowers to very high ethylene concentrations (50 microliters per liter) for 48 hours did not promote corolla abscission or senescence. 1-Aminocyclopropane-1-carboxylic acid, the immediate precursor of ethylene, increased ethylene production by unpollinated flowers more than 100-fold, but did not promote corolla abscission. 1-Aminocyclopropane-1-carboxylic acid did enhance corolla abscission of pollinated flowers. It is concluded that the main effect of pollination in inducing corolla abscission of cyclamen is by rendering the tissue sensitive to ethylene, apart from the promotion of ethylene production.     

Style-controlled wilting of the flower

Differences in rate of wilting in cross-, self-and unpollinated flowers of self-incompatiblePetunia hybrida L. clone W166H appeared to be significant. Wilting rate was fastest following cross-pollination and slowest in unpollinated flowers. The difference between wilting behaviour of cross- and self-pollinated flowers was not caused by rate of pollen tube growth and not by the incompatibility (recognition or rejection) reaction either. It is assumed, that, following pollination, the wilting reaction is only retarded after penetration of pollen tubes of the same genetic composition as the style (complete self-pollination). The number of viable pollen grains necessary to initiate a maximal wilting-rate of flowers following cross- and self-pollination is about 800, which means that a fifth of the stigmatic surface must be covered with living pollen grains. It is suggested that pollen tube penetration and injury of the style have a similar influence on the initiation of wilting.Wilting-rate following pollination is faster in young plants as compared with wilting in old plants. The wilting process of unpollinated and self-pollinated flowers started in the early morning and lasted till afternoon. Cross-pollinated flowers wilted independently of the hour of the day. The role of flower-wilting as a means of communication to the environment with regard to pollination of the style is discussed.     

Transient Water Stress in Carnation Flowers: Effect of Amino-oxyacetic Acid

A short and temporary water stress imposed on cut carnationflowers (Dianthus caryophyllus L., cv. White Sim) flowers advancedsenescence symptoms, including ethylene production and wilting.Pretreatment with amino-oxyacetic acid (AOA) resulted in anincrease of the resistance of the flowers to water stress: waterloss during stress was reduced, recovery was more rapid andwilting was delayed. Water stress accelerated the decrease inlevel of membrane phospholipids, but pretreatment with AOA counteractedthis effect. Since the content of membrane sterols was not affectedby the treatments, the mole ratio of sterol to phospholipidincreased in water-stressed flower petals but not in stressedflowers pretreated with AOA. Membrane permeability and fluiditywere also adversely affected by water stress and AOA: waterstress alone resulted in an increase in permeability and a decreasein fluidity, but in AOA-pretreated stressed flower petals theseparameters were similar to those of nonstressed control flowerpetals. On the basis of these results two main conclusions can be drawn:(a) Water stress induces alterations in the physical and compositionalproperties of carnation petal membranes, (b) Pretreatment ofthe flowers with AOA influences petal membrane traits, mostprobably via modifications in phospholipid turnover, in a waywhich counteracts the effects of water stress. Key words: Amino-oxyacetic acid, Water stress, Carnation flowers     

Ethylene biosynthetic genes are differentially expressed during carnation (Dianthus caryophyllus L.) flower senescence

Ethylene production and expression patterns of an 1-aminocyclopropane-1-carboxylic acid (ACC) oxidase (CARAO1) and of two ACC synthase (EC 4.4.1.14) genes (CARACC3 and CARAS1) were studied in floral organs of cut carnation flowers (Dianthus caryophyllus L.) cv. White Sim. During the vase life and after treatment of fresh flowers with ethylene, production of ethylene and expression of ethylene biosynthetic genes first started in the ovary followed by the styles and the petals. ACC oxidase was expressed in all the floral organs whereas, during the vase life, tissue-specific expression of the two ACC synthase genes was observed. After treatment with a high ethylene concentration, tissue specificity of the two ACC synthase genes was lost and only a temporal difference in expression remained. In styles, poor correlation between ethylene production and ACC synthase (CARAS1) gene expression was observed suggesting that either activity is regulated at the translational level or that the CARAS1 gene product requires an additional factor for activity.Isolated petals showed no increase in ethylene production and expression of ethylene biosynthetic genes when excised from the flower before the increase in petal ethylene production (before day 7); showed rapid cessation of ethylene production and gene expression when excised during the early phase of petal ethylene production (day 7) and showed a pattern of ethylene production and gene expression similar to the pattern observed in the attached petals when isolated at day 8. The interorgan regulation of gene expression and ethylene as a signal molecule in flower senescence are discussed.     

Ethylene Co-ordinates Petal Abscission in Red Raspberry (Rubus idaeus L.) Flowers

The time-course of flower development of Rubus idaeus L. cv.Glen Clova was studied on detached buds opened in the laboratory.After sepal and petal opening petal abscission occurred withthe petals from an individual flower being shed over 3-4 h.Abscission was accompanied by a peak in ethylene production.Treatment of flowers with aminoethoxyvinylglycine eliminatedthe peak in ethylene production but did not prevent petal abscission.However, petal loss was much slower, taking place over a periodof days rather than hours. Abscission was more effectively retardedby silver thiosulphate. Exogenous ethylene accelerated the rateof petal abscission and senescence. The increase in ethyleneproduction coincident with petal abscission appears to accelerateand co-ordinate the shedding of the separate petals on an individualflower. If ethylene is important in the induction of abscissionit would appear that the low rate of production sustained inthe presence of aminoethoxyvinylglycine must be sufficient.Copyright1993, 1999 Academic Press Rubus idaeus L., raspberry, flower, petal, abscission, ethylene     

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     

Pollination-Induced Corolla Wilting in Petunia hybrida Rapid Transfer through the Style of a Wilting-Inducing Substance

Pollination or wounding of the stigma of Petunia hybrida flowers led to the generation of a wilting factor and its transfer to the corolla within 4 hours. This was concluded from the effects of time course removal of whole styles. In this 4-hour period, pollen tubes traversed only a fraction of the total distance to the ovaries. Both pollination and wounding of the stigma immediately resulted in an increase of ethylene evolution. Accelerated wilting, however, occured only when treated styles remained connected with the ovaries, and not when they were detached and left in the flower. A wilting factor was found in eluates collected from the ovarian end of the styles, only in the case of previous pollination or wounding. In such eluates, the level of the ethylene precursor 1-amino-cyclopropane-1-carboxylic acid was below detection.

These observations suggest a material nature of the wilting factor in Petunia flowers, which rapidly passes through the style to the corolla, but which is different from 1-aminocyclopropane-1-carboxylic acid.

     

Pollination-induced flower senescence: a review

Ethylene has long been implicated in the control of the senescence of many cut flower species, but the control of senescence in relation to wild species has received much less attention. The longevity of individual flowers varies greatly from species to species; in some each flower is open for just a few hours, whilst in others the flower may persist for several weeks, or even months. The functional life of the flower may be terminated by petal wilting, abscission or a colour change of all, or part, of the perianth. In some species pollination appears to reduce floral longevity whilst in others, particularly those species having short-lived flowers, the pattern of flower development and senescence appears unaffected by pollination.Examples of the various pollination-induced strategies shown by plants are presented and the role of ethylene and other potential mediators of senescence in these processes discussed.