Spatial and temporal distribution of mineral nutrients and sugars throughout the lifespan of <Emphasis Type="Italic">Hibiscus rosa-sinensis</Emphasis> L. flower

Although the physiological and molecular mechanisms of flower development and senescence have been extensively investigated, a whole-flower partitioning study of mineral concentrations has not been carried out. In this work, the distribution of sucrose, total reducing sugars, dry and fresh weight and macro and micronutrients were analysed in Hibiscus rosa-sinensis L. petals, stylestigma including stamens and ovary at different developmental stages (bud, open and senescent flowers). Total reducing sugars showed the highest value in petals of bud flowers, then fell during the later stages of flower development whereas sucrose showed the highest value in petals of senescent flowers. In petals, nitrogen and phosphorus content increased during flower opening, then nitrogen level decreased in senescent flowers. The calcium, phosphorus and boron concentrations were highest in ovary tissues whatever the developmental stage. Overall, the data presented suggests that the high level of total reducing sugars prior the onset of flower opening contributes to support petal cells expansion, while the high amount of sucrose at the time of petal wilting may be viewed as a result of senescence. Furthermore, this study discusses how the accumulation of particular mineral nutrients can be considered in a tissue specific manner for the activation of processes directly connected with reproduction.     

Sites of ethylene production in the pollinated and unpollinated senescing carnation (Dianthus caryophyllus) inflorescence

Production of endogenous ethylene from the styles, ovary and petals of pollinated and unpollinated flowers of Dianthus caryophyllus L. was measured. The rate of ethylene production of cut, unpollinated flowers aged in water at 18°C was low until the onset of petal wilting, when a rapid surge of ethylene occurred in all tissues. The flower ethylene production was evolved mostly from the styles and petals. The bases of petals from unpollinated, senescing flowers evolved ethylene faster and sometimes earlier than the upper parts. Treatment of cut flowers with propylene, an ethylene analogue, accelerated wilting of flower petals and promoted endogenous ethylene production in all flower tissues. Pollination of intact flowers also promoted endogenous ethylene production and caused accelerated petal wilting within 2–3 days from pollination. Although the data are consistent with the hypothesis that ethylene forms a link between pollination of the style and petal wilting, in the unpollinated flower the style and petals can evolve a surge of ethylene independently of each other, about the time when the petals irreversibly wilt. The results are discussed in relation to the role of ethylene in flower senescence.     

An Effect of Ethylene on the Distribution of 14C-sucrose from the Petals to Other Flower Parts in the Senescent Cut Inflorescence of Dianthus caryophyllus

The distribution of carbon-14 in the flower parts of the cutcarnation inflorescence after feeding ¹⁴C-sucrose through thepetals was studied during natural ageing and after ethylenetreatment. Levels of ethylene which caused irreversible wiltingof petals also promoted an accelerated transfer of the radioactivesucrose to the nectar, gynaecium and stem. Since the nectarreceived a relatively large proportion of the radioactive carbon,the composition of the sugars in the nectar and the vascularizationof the nectary were investigated. Sucrose comprised about 85per cent of the nectar sugars and the balance was glucose andfructose. The vascular tissue closest to the nectary consistedof phloem elements; tracheary elements terminated deeper inthe receptacle and were surrounded by a ring of phloem. Thepercentage of solutes in the nectar was about 18 per cent andincreased when the flowering stems were placed in sucrose solutions;the solutes in the nectar were principally sugars. Taken togetherthe results show that the nectary can act as a sink for sucroseand, in the flower at least, that translocation of sucrose takesplace in the phloem. The results provide further evidence forthe hypothesis that ethylene promotes mobilization of substrateand an efflux of material from petals to the gynaecium, nectarand stem.     

Senescence of cut carnation flowers: Ovary development and CO2 fixation

In the presence of ethylene, which enhances carnation flower senescence, carbohydrates contribute to ovary growth not only from the stem and calyx but also from the petals. With silver thiosulphate and ethanol treatments which delay flower senescence, the petals remain the active sink and ovary development is suppressed. Ethylene stimulated chloroplast development in the ovary wall. However, the calyx plus stem of all treatments showed the greater photosynthetic ability and transported a major portion of the synthesised products to the ovary.     

Contribution of sepals and gibberellin treatments to growth and development of rose (Rosa hybrida) flowers

The fresh weight of sepals during the development of the rose flowerbuds from 4 mm to 22 mm in diameter increased fromabout 30 mg to ca. 350 mg. However, due to a morerapid gain in the total fresh weight of the flower, the sepal fresh weight as aproportion of the total weight of the buds decreased from about 55% to only 8%at the end of the measurement period. The net photosynthesis of sepals,measuredclose to the flower harvest, was approximately 60% of that in the youngest,uppermost leaves whereas no photosynthesis occured in the petals. Theconcentration of sucrose in petals of almost fully developed, desepalledflowerswas 15% lower in comparison with the control flowers with intact sepals. On theother hand, the concentration of sucrose in petals of control and desepalledflowers that were kept for 10 days in complete darkness was equal, reachingabout 50% of the concentration in petals of flowers grown in the light.Periodicmeasurements of reducing sugars in the petals did not show differences in theirconcentration between the control and desepalled flowers during the first 8daysafter sepal removal. After an additional four days the concentration ofreducingsugars in petals of the desepalled flowers was only 50% in comparison to thatinpetals of control flowers. Excising the sepals reduced fresh and dry weights,aswell as the length of buds and the peduncles, indicating that sepals may be asource of gibberellins during flower development. Treatment with50mg GA₃ in lanolin paste, completely restored thelength of the peduncles, but only partially restored the other measuredparameters of the flowers. Formation of 'star-shape' abnormality indesepalled flowers, which is a common phenomenon in rose flowers exposed toexternal ethylene was completelly prevented by applying GA₃ afterthesepals were excisied. This supported the previously suggested hypothesis aboutthe flinction of gibberellins in reducing the sensitivity of rose flower organsto ethylene.     

Effects of Ethylene and Sucrose on Translocation of Dry Matter and 14C-Sucrose in the Cut Flower of the Glasshouse Carnation (Dianthus caryophyllus) During Senescence

The translocation and distribution of dry matter were studiedin the floral and vegetative parts of the cut carnation duringsenescence. The change in dry weights of the tissues and theamount of radioactivity recovered from them after feeding with¹⁴C-sucrose were measured. Treatments with ethylene and sucrosewere used to alter the rate of senescence of the flowers. Sucrosemoved through the stem relatively unchanged but was rapidlyinverted and metabolized in the petals. During natural ageing,¹⁴C moved from the stem to the flower and the movement was enhancedby exogenous sucrose, which also reduced the loss of dry matterin the petals and promoted their growth. Treatment with ethylenecaused petals to wilt and lose dry weight, and ovaries to enlargeand increase in dry weight. The distribution of radioactivityin flowers fed with 14C-sucrose before and after ethylene treatmentsupported the observation that dry matter was translocated betweenthe flower parts. The results indicate that a change in thesource-ink relationships of the flower parts contributes tothe factors that determine the rate of flower senescence.     

Interrelationship between the Different Flower Parts during Emasculation-Induced Senescence in Cymbidium Flowers

In Cymbidium flowers emasculation by removal of the anther capand the pollinia, led to rapid colouration of the lip and advancedwilting of the petals and sepals. The ethylene production ofwhole flowers showed an emasculation-induced early peak in ethyleneevolution followed some days later by a second increase concomitantwith the wilting of the flower. In non-emasculated flowers theethylene production increased later and simultaneously withcolouration of the lip and wilting of the petals and sepals.At all stages of senescence, the contribution of the lip, petals,and sepals to the total amount of ethylene produced was negligible. Parallel to the increase in ethylene production of whole flowers,an increase in 1-aminocyclopropane-l-carboxylic acid (ACC) andmalonyl-ACC (MACC) in the central column and, to a lesser extent,in the ovary was observed. Also an increase in internal ethyleneconcentration was demonstrated and this, in contrast, was apparentin all the different flower parts. The activity of the ethylene-formingenzyme in lips, petals, and sepals showed an increase afteremasculation and such an effect could also be induced by treatmentof isolated lips with low concentrations of ethylene. The data indicate that senescence in Cymbidium flowers is regulatedby the central column and perhaps the ovary and that both ACCand ethylene may play a signalling role in inter-organ communication. Key words: 1-aminocyclopropane-l-carboxylic acid, ethylene, Cymbidium, senescence     

Changes of Carbohydrates in Pepper (Capsicum annuum L.) Flowers in Relation to Their Abscission Under Different Shading Regimes

Abscission of pepper flowers is enhanced under conditions oflow light and high temperature. Our study shows that pepperflowers accumulate assimilates, particularly in the ovary, duringthe day time, and accumulate starch, which is then metabolizedin the subsequent dark period. With the exception of the petals,the ovary contains the highest total amounts of sugars and starch,compared with other flower parts and contains the highest totalactivity, as well as activity calculated on fresh mass basis,of sucrose synthase, in accordance with the role of this enzymein starch biosynthesis. Low light intensity or leaf removaldecreased sugar accumulation in the flower and subsequentlycaused flower abscission. The threshold of light intensity fordaily sugar accumulation in the sink leaves was much lower thanin flowers, resulting in higher daytime accumulation of sugarsin the sink leaves than in the adjacent flower buds under anylight intensity, suggesting a competition for assimilates betweenthese organs. Flowers of bell pepper cv. ‘Maor’and ‘899’ (sensitive to abscission) accumulatedless soluble sugars and starch under shade than the flowersof bell pepper cv. ‘Mazurka’ and of paprika cv.‘Lehava’ (less sensitive). The results suggest thatthe flower capacity to accumulate sugars and starch during theday is an important factor in determining flower retention andfruit set. Pepper; Capsicum annuum L.; abscission; shading; pepper flowers; ovary; leaves; sugars; starch; acid invertase; sucrose synthase     

Endogenous levels of abscisic acid in aging carnation flower parts

Aging carnation flower parts were used to determine whether or not any correlation existed between the concentration of abscisic acid (ABA) and a predisposition of the tissue for ethylene synthesis. Levels of ABA were measured using an enzyme-linked immunosorbent assay (ELISA) following purification steps including prepacked silica gel columns. Increased ABA levels paralleled the increase of ethylene and the onset of irreversible wilting in the carnation petals. Neither the green tissue nor the receptacle showed any sign of wilting with the remainder of the flower parts, but increased ABA was detected in both tissues subsequent to, or coincident with, the ethylene climacteric peak in the senescing petals. An increase of ABA in both the styles and the ovary was detected in the preclimacteric flower, and did not appear to be triggered by the production of ethylene. Increased ABA in the gynoecium also did not result in the onset of ethylene production in the preclimacteric flower.     

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.     

Translocation of 14C, carbohydrate content and activity of the enzymes of sucrose metabolism in rose petals temperatures

Both export of ¹⁴C from the source leaves of roses (Rosa × hybrida cv. Golden Times) and import of ¹⁴C to the petals were reduced by plant exposure to low night temperature. However, the import was affected to a greater extent than the export. During all stages of flower bud development the concentration of reducing sugars in petals of roses grown at reduced night temperature was lower than in petals of plants grown at higher night temperature. There was no significant difference in starch content in response to the night temperature, and the content of starch decreased toward complete flower bud opening. The concentration of sucrose in flowers at the low night temperature remained low during all stages of flower bud development, while at the high night temperature the concentration of sucrose increased during flower bud development, reaching a peak at the stage when petals start to unfold. At both temperatures the concentration of sucrose declined at complete flower opening. The activity of sucrose synthase (EC 2.4.1.14) was inhibited by low temperature in young rose shoots more than in the petals, while the activity of acid invertase (EC 3.2.1.26) was affected similarly in both tissues by the temperature treatments.     

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.     

Nonosmotic inhibition by sugars of the ethylene-forming activity associated with microsomal membranes from carnation petals

The activity of the membrane-bound ethylene-forming enzyme, previously reported in carnation (Dianthus caryophyllus L. cv White Sim) petals (Mayak, Legge, Thompson 1981 Planta 153: 49-55), is inhibited by sugars. Of the various sugars tested, sorbitol was the most effective and glucose the least. The effect of sugars was also evaluated on solubilized ethylene-forming enzyme activity, obtained by the use of 0.6% Nonidet NP-40 detergent. Similar to the membrane-bound activity, the solubilized activity was also inhibited. Kinetic studies revealed that the inhibition by sugars is reversible, and that inhibition by sucrose is uncompetitive while that by sorbitol is competitive. During senescence of petals, a decline in sugar content and climacteric like increase in ethylene occurs. Hence, the physiological relevance of sugar inhibition and its possible involvement in the regulation of ethylene biosynthesis is suggested.     

Effects of Sucrose on Water Relations of Cut, Senescing, Carnation Flowers

Carnation flower stems were stood in water or sucrose solutionand changes in water content, water and osmotic potential, turgorpressure and solutes (sugars, nitrogen, phosphorus, potassium)of petals were measured throughout the flower life. In bothtreatments the petals had a higher specific water content atincipient wilting than when the flowers were first cut. In water,turgor pressure decreased rapidly after the seventh day becauseof a decrease in tissue solute content. In sucrose solution,loss, of solutes was delayed probably because the sugar provideda respiratory substrate to maintain cell membrane integrity.In these cells, sugars and water accumulated causing decreasesin water potential and osmotic potential. Solutes and waterwere lost at about day 15 and turgor pressure decreased. Therewas some evidence that from about day 11 cells were so gorgedwith sugars that they burst when they were placed in water duringthe adjustment of water content prior to water potential measurements. Most of the initial petal osmotic energy content could be accountedfor by sugar, potassium, and anions associated with potassium,but in water, as the petals aged and sugar content decreased,so the potassium ions contributed a larger proportion of theosmotic energy; with stems in sucrose, the endogenous sugarcontent (reducing sugars plus sucrose) contributed an increasingproportion of the total osmotic energy. Dianthus caryophyllus, carnation, flowers, water relations, senescence     

Sucrose metabolism and IAA and ethylene production in muskmelon ovaries

Changes induced by the pollination of ovaries may be mediated by phytohormones and involve sudar-mediated by phytohormones and involve sugar-metabolizing enzymes. In order to further explore these relationships, soluble sugars, sucrose-phosphate synthase (EC 2.4.1.14), sucrose synthase (EC 2.4.1.13), acid and neutral invertases (EC 3.2.1.26), indole-3-acetic acid (IAA), and ethylene were investigated in muskmelon (Cucumis melo L.) ovaries sampled before, during, and after anthesis. The fresh weight of ovaries increased 100% within 48 h after pollination, but did not change significantly in the absence of pollination. While sugar content per ovary increased after pollination, sugar content per mg protein was unaffected. Sucrose was not detected in nonpollinated ovaries 48 h after anthesis. Free IAA content was highest in ovaries sampled 48h before anthesis. Pollination had no immediate effect on IAA content per mg protein in postanthesis ovaries. Although detected in all ovaries sampled, ethylene production increased significantly only in nonpollinated ovaries. Activity of sucrose-phosphate synthase was the same at all stages. The specific activities of sucrose synthase and the invertases were highest in nonpollinated ovaries. The increase in rate of sugar import into ovaries following pollination was not accompanied by an increase in the specific activity of any enzyme assayed, but was coincident with an increase in the total activity per ovary of surcose synthase and acid invertase. There appears to be no direct relationship between sucrose-metabolizing enzymes, IAA or ethylene in developing pollinated ovaries but the increase in sucrose cleavage activity in nonpollinated ovaries may be related to the increase in ethylene production.     

Endogenous ethylene does not regulate opening of unstressed Iris flowers but strongly inhibits it in water-stressed flowers

The floral buds of Iris flowers (Iris x hollandica) are enclosed by two sheath leaves. Flower opening depends on lifting the flower up to a position whereby the tepals can move laterally. This upward movement is carried out by elongation of the subtending pedicel and ovary. In the pedicels and ovaries of unstressed control flowers, the concentration of ACC (1-aminocyclopropane-1-carboxylic acid) and the rate of ethylene production increased during d 0-1 of flower opening, and then decreased. Exposure to ≥200nLL(-1) ethylene for 24h at 20°C inhibited elongation of the pedicel+ovary, and inhibited flower opening. However, pulsing of unstressed flowers with solutions containing inhibitors of ethylene synthesis (AOA, AVG), or an inhibitor of ethylene action (STS), did not affect pedicel+ovary elongation or flower opening. When the flowers were dehydrated for 2 d at 20°C and 60% RH, they did not open when subsequently placed in water, and showed inhibited elongation in the pedicel+ovary. This dehydration treatment resulted in elevated pedicel+ovary ACC levels and in increased ethylene production. Treatment with STS prevented the increase in ACC levels and ethylene production, overcame the effect of dehydration on elongation of the pedicel+ovary, and resulted in full flower opening. It is concluded that flower opening in unstressed Iris flowers is not regulated by endogenous ethylene. An increase in endogenous ethylene above normal levels during stress, by contrast, strongly inhibited flower opening, due to its inhibitory effect on elongation of the pedicel+ovary.     

Combined Effects of Abscisic Acid and Sucrose on Growth and Senescence of Rose Flowers

The effects of sucrose and abscisic acid (ABA) and their interaction on development and senescence of petals were studied with leafless roses cultivar Super Star. Sucrose and ABA had opposing effects on the cut flowers. Sucrose retarded and ABA promoted processes associated with senescence: wilting, increase in pH, “blueing” and decrease in protein content of petals. These opposing effects are mutually antagonized when both chemicals are applied. ABA applied to flowers cut at the bud stage, promoted the rate of petal growth (but not their final size), increased respiration and caused a decrease in sucrose and an increase in level of reducing sugars. It is suggested that one way by which ABA accelerates senescence of cut roses is by promoting petal growth and respiration, thus decreasing the carbohydrate level in the petals and triggering the chain of metabolic processes leading to aging.     

The Relationship between Sucrose Supply, Sucrose-cleaving Enzymes and Flower Abortion in Pepper

Abortion of pepper flowers depends on the light intensity perceivedby the plant and on the amounts of sucrose taken up by the flower(Aloni B, Karni L, Zaidman Z, Schaffer AA. 1996.Annals of Botany78: 163–168). We hypothesize that changes in the activityof sucrose-cleaving enzymes within the flower ovary might beresponsible for the changes in flower abortion under differentlight conditions. In the present study we report that the activityof sucrose synthase, but not of cytosolic acid invertase, increasesin flowers of pepper plants which were exposed, for 2 d, toincreasing photosynthetically active radiation (PAR) in therange of 85–400 µmol m^-2s^-1at midday. Sucrose synthaseactivity increased in parallel with the increasing concentrationsof starch in the flower ovary. Feeding flower explants, preparedfrom 3-d-predarkened plants, with 100 mM sucrose for 24 h, causeda 23% increase in reducing sugars and a 2.5-fold increase instarch concentration, compared with explants fed with buffer.Likewise, feeding explants of pepper flowers with sucrose, glucose,fructose and also mannitol increased the sucrose synthase activityin the ovaries. Concomitantly, sucrose, glucose and fructose,but not mannitol, reduced the abortion of flower explants. Itis suggested that sucrose entry into the flower increases theflower sink activity by inhibiting abscission and inducing metabolicchanges, thus enhancing flower set. Pepper; Capsicum annuum L.; abscission; light; pepper flowers; sucrose; glucose; fructose; starch; acid invertase; sucrose synthase