Inhibition of ethylene responses by 1-Methylcyclopropene and 3-Methylcyclopropene

3-Methylcyclopropene (3-MCP) binds to the ethylene receptor and blocks it for several days, but concentrationswise is less effective than 1-methylcyclopropene (1-MCP). In diverse ethylene-responsive systems, including ripening of mature-green bananas (Musa sapientum L.), inhibition of growth in etiolated pea (Pisum sativum L.) seedlings, abscission of orange (Citrus sinensis L.) leaf explants and mung bean (Vigna radiata L.) leaves, and wilting of campanula (Campanula carpatica) and kalanchoe (Kalanchoë blossfeldiana) florets, full inhibition of the ethylene response required higher concentrations of 3-MCP. Depending on the experimental system, the effective concentration of 3-MCP was from 5 to 10 times higher than that required for 1-MCP.     

Fruit age and changes in abscisic Acid content, ethylene production, and abscission rate of cotton fruits

The relationships of fruit age, abscisic acid (ABA) concentration, ethylene evolution, and abscission rates were studied in an effort to determine why cotton (Gossypium hirsutum L., cv. Deltapine 16) fruits rarely abscise more than 15 days after anthesis. Because abscission of cotton fruits is increased by conditions that limit photosynthesis, greenhouse-grown plants with fruits of various ages were placed in dim light for 3 days to induce high rates of fruit abscission. Abscission rates, ABA concentrations, and ethylene evolution rates were determined for fruits of various ages. Almost all of the young fruits abscised, but abscission rate declined with age until almost no abscission was observed in fruits that were 15 or more days past anthesis.     

The roles of abscisic acid and ethylene in the abscission and senescence of cocoa flowers

Cocoa flowers have a limited period of longevity; more than 90% of unpollinated flowers abscised within 32 h after anthesis. Abscisic acid (ABA) levels increased significantly prior to abscission. By 21 h after anthesis, ABA levels had increased almost 10-fold, and by 32 h flowers had 20-fold higher levels of ABA than at anthesis. Fluridone completely inhibited both the increase in ABA, the formation of an abscission zone, and the abscission and senescence of flowers. In contrast, ethylene production increased only slightly 21 h after anthesis and was only 2-fold higher after 32 h. Aminoethoxyvinylglycine (AVG) delayed but did not prevent abscission. In cocoa flowers, ABA is the primary regulator of abscission; ethylene accelerates abscission but only in the presence of ABA. Naphthalene acetic acid (NAA) treatment of flowers at anthesis prevented abscission zone formation and flower abscission, but did not induce fruit set. All parts of the NAA-treated flower except the pedicel senesced after 6 days. NAA+AVG treatment only delayed, whereas fluridone treatment completely prevented flower senescence.     

Ethylene Evolution following Treatment with 1-Aminocyclopropane-1-carboxylic Acid and Ethephon in an in Vitro Olive Shoot System in Relation to Leaf Abscission

1-Aminocyclopropane-1-carboxylic acid (ACC) supplied via the cut base of detached olive shoots caused a burst of ethylene from leaves, but other cyclopropanes tested did not exhibit this effect. Ethephon (ET) and another ethylene-releasing compound caused a prolonged increase in ethylene evolution. ACC had only a very limited effect on leaf abscission regardless of concentration, whereas shoots placed with cut bases in ET for 60 to 80 minutes exhibited 100% leaf abscission within 90 hours. Shoots with inflorescences treated with ET just prior to anthesis began to wilt in vitro within 20 to 30 hours and failed to exhibit leaf abscission. At earlier stages of development, ET induced more leaf abscission on reproductive shoots than on vegetative shoots. It is suggested that the duration of ethylene evolution from the leaves governs their potential for abscission and that bursts of ethylene evolution even though large in amount may not induce abscission.     

Changes in Abscisic Acid and Indoleacetic Acid before and after Anthesis Relative to Changes in Abscission Rates of Cotton Fruiting Forms

Cotton (Gossypium hirsutum L.) fruiting forms exhibit pronounced changes, with age, in their probability of abscission. Large floral buds rarely abscise, but after anthesis the young fruits (bolls) have a high probability of abscising. Abscission rate reaches a peak about 5 to 6 days after anthesis and then gradually decreases. An experiment was conducted to try to determine the reason for the rapid and pronounced increase in probability of abscission just after anthesis. Cotton was grown in the field and fruiting forms of various ages from 9 days before to 9 days after anthesis were all harvested the same day and subsequently analyzed for ABA and IAA. The concentration of ABA decreased slightly at anthesis and increased gradually thereafter. In contrast, the concentration of IAA was high before anthesis and then decreased at anthesis to about one-fifth the previous concentration. IAA remained low for at least 4 days after anthesis and then increased rapidly between 7 and 9 days after anthesis. The high concentration of IAA in floral buds before anthesis is probably a major factor in their resistance to abscission. Likewise, the low concentration of IAA at anthesis and for about 4 days thereafter may promote fruit abscission during the young boll stage.     

Prevention of floret abscission for Agapanthus praecox requires an adequate supply of carbohydrate to the developing florets

Vase solutions of 2.5 or 5% (w/v) sucrose markedly reduced the abscission of all open florets and buds on cut Agapanthus praecox stems. Pulsing cut stems with these solutions was not as effective as continuous treatments. Pulse treatments with 10% sucrose for 4–24 h were relatively ineffective at reducing floret and bud abscission but longer pulses of 48 h reduced abscission. The relative ineffectiveness of sucrose pulse treatments appeared to be due to the low water uptake of the stems (1–3 ml/day). Reducing the number of florets and buds on an inflorescence reduced abscission of florets and buds, and increasing stem length from 25 cm to 50 cm decreased bud abscission. This reduction in abscission is possibly due to the increased availability of assimilates for the remaining buds and florets or reduced competition for assimilates. Exogenous ethylene treatments (9 µl/L for up to 24 h) had no effect on abscission, although STS treatment (4 mM, 4 h) significantly reduced floret abscission when stems were held in vase solutions of sucrose. We conclude that postharvest floret abscission in A. praecox is influenced primarily by the availability of assimilates to the developing florets.     

Nucellar Senescence and Ethylene Production as they Relate to Avocado Fruitlet Abscission

Events preliminary to avocado (Persea americana Mill) fruitletabscission include senescence of the nucellus and seed coat.The dynamics of nucellar deterioration and ethylene productionleading to seed abortion and abscission in avocado was examined.Excised branches bearing clusters of fruit from 1.0–2.5cm diameter were placed in humid chambers to reduce transpirationalwater loss. Fruitlets synchronously began nucellar and seedcoat deterioration 27–33 h after excision and rapidlyprogressed through stages of increasing degradation culminatingin abscission approximately 2 days later. The nucellus-seedcoat produced a temporary burst of ethylene at the first visiblesign of nucellar senescence followed by less ethylene productionin the mesocarp approximately 12 h later. All fruit underwentnucellar degradation prior to abscission. Exogenously appliedethylene accelerated fruitlet abscission with concentrationsas low as 1.0µ 1^–1 and with maximum response at100µl^–1 or greater. Maximal response took 2 days.Aminoethoxyvinyl-glycine (AVG) at 30 µ M inhibited ethyleneproduction and fruitlet abscission. The senescence process,however, was not af fected in any way by ethylene or AVG treatments.Observations of attached fruit suggest that nucellar-seed coatsenescence, concomitant ethylene production, and resulting abscissiontake place in a manner and within a time period similar to thatobserved on detached branches. It is concluded that nucellarand seed coat senescence is prerequisite to avocado fruitletabscission, and the time required from the first indicationof nucellar breakdown to abscission of that fruitlet appearsto be approximately 2 days. The senescence process is responsiblefor a large, transient rate increase in ethylene productionby the nucellus and perhaps seed coat. Ethylene is consideredto be the result rather than the cause of nucellar-seed coatsenescence. The ethylene thus produced induces fruit abscission.     

The never ripe mutation blocks ethylene perception in tomato.

Seedlings of tomato fruit ripening mutants were screened for their ability to respond to ethylene. Ethylene induced the triple response in etiolated hypocotyls of all tomato ripening mutants tested except for one, Never ripe (Nr). Our results indicated that the lack of ripening in this mutant is caused by ethylene insensitivity. Segregation analysis indicated that Nr-associated ethylene insensitivity is a single codominant trait and is pleiotropic, blocking senescence and abscission of flowers and the epinastic response of petioles. In normal tomato flowers, petal abscission and senescence occur 4 to 5 days after the flower opens and precede fruit expansion. If fertilization does not occur, pedicel abscission occurs 5 to 8 days after petal senescence. If unfertilized, Nr flowers remained attached to the plant indefinitely, and petals remained viable and turgid more than four times longer than their normal counterparts. Fruit development in Nr plants was not preceded by petal senescence; petals and anthers remained attached until they were physically displaced by the expanding ovary. Analysis of engineered 1-aminocyclopropane-1-carboxylate (ACC) synthase-overexpressing plants indicated that they are phenotypic opposites of Nr plants. Constitutive expression of ACC synthase in tomato plants resulted in high rates of ethylene production by many tissues of the plant and induced petiole epinasty and premature senescence and abscission of flowers, usually before anthesis. There were no obvious effects on senescence in leaves of ACC synthase overexpressers, suggesting that although ethylene may be important, it is not sufficient to cause tomato leaf senescence; other signals are clearly involved.     

Chilling-induced leaf abscission of Ixora coccinea plants. I. Induction by oxidative stress via increased sensitivity to ethylene

Exposing ixora ( Ixora coccinea ) plants to chilling temperatures (3–9°C for 3 days) resulted in increased leaf abscission, initiated 3 days after transfer to 20°C. Exposure to chilling also induced a 7-fold increase in ethylene production rates of abscission zone (AZ) tissue during the initial 5 h after chilling. The ethylene burst resulted from the high levels of 1-aminocyclopropane-1-carboxylic acid (ACC) accumulated in the AZ during the chilling period. ACC levels following chilling decreased also due to enhanced conjugation to 1-(malonylamino)cyclopropane-1-carboxylic acid (MACC). Treating plants prior to chilling with antioxidants, such as butylated hydroxyanisole (BHA), n -propyl gallate (PG), and vitamin E, significantly reduced chilling-induced leaf abscission. This effect was obtained despite the fact that ethylene production in the treated plants resembled that of chilled plants receiving no BHA. In addition, exposure of plants to ethylene (0.5–10 μl l⁻¹) for 1–3 days significantly enhanced leaf abscission only when they had been pre-chilled. These data imply that chilling-induced leaf abscission was closely correlated with increased sensitivity of the AZ to ethylene rather than with the chilling-induced ethylene burst. Based on the findings that the ethylene action inhibitor, 1-methylcyclopropene (1-MCP), and the antioxidant BHA inhibited both the chilling-induced and the ethylene-enhanced leaf abscission, it is concluded that: (1) although ethylene is essential for chilling-induced abscission, it is not the triggering factor; (2) oxidative processes derived from the chilling stress seem to be the trigger of chilling-induced leaf abscission, operating via increased sensitivity to ethylene.     

Early Application of Ethrel Extends Tomato Fruit Cell Division and Increases Fruit Size and Yield with Ripening Delay

Flowers of tomato (Lycopersicon esculentum Mill.) plants cv. Castle Rock were sprayed with 100 ppm of ethrel, 0.5 mm aminooxyacetic acid (AOA), or water (control) 2 days after anthesis. The fruit period of cell division was extended up to 16–18 days after anthesis with the application of ethrel but reduced from 10–12 days (control) down to only 6–8 days with the application of AOA. In a trend opposite to AOA application, fruits that received ethrel treatment were of higher ethylene and 1-aminocyclopropane-1-carboxylic acid (ACC) levels than control. This was noticed not only during the first 2 weeks after anthesis but also during the fruit climacteric phase. Mesocarp cells of ethrel-treated fruits were greater in number/mm² but smaller in size than control; an opposite trend was obtained with the application of AOA. This was observed for a period of 18 days after anthesis, but by that time or at earlier ages, fruits of AOA treatment were larger in size and heavier in weight than control, and both were larger and heavier than ethrel-treated ones. At 5 weeks after anthesis and thereafter, the fruit response to all treatments was totally reversed because early ethrel-treated fruits became significantly larger in size and heavier in weight with a ripening delay of about 10 and 15 days compared with those of control and AOA-treated ones, respectively. When the same treatments were applied to the whole plant, similar results were obtained because the early application of ethrel increased the fruit yield by about 15% over control with a pronounced ripening delay; an opposite trend was obtained with the application of AOA. No significant differences were found among all treatments in terms of flower or fruit abscission or fruit number/plant. The data suggest that ethylene regulates tomato fruit transmission from cell division to cell enlargement. In addition, fruit cell division is terminated only when endogenous ethylene decreases to its basal level, allowing cell enlargement to dominate and proceed as in the case of the early application of AOA. The ripening delay of ethrel-treated fruits may be caused by the longer time required for the increased cell number to reach maturation. A low level of ethrel application at the tomato early fruiting stage may be used for increasing fruit yield by increasing fruit size and consequently its quality. Received June 1, 1998; accepted December 7, 1998     

Ethylene, a regulator of young fruit abscission

In an earlier study we reported that detached cotton flowers produced sufficient ethylene before the period of natural abscission to suggest that ethylene might be a natural regulator of young fruit abscission. The present report explores this probability further. Intact cotton (Gossypium hirsutum L.) fruits produced ethylene at rates as high as 36 μl ethylene/kg fresh wt·hr during the 2 days before they abscised. Direct measurements of ethylene in gas samples withdrawn from fruits indicated that production of 1 μl ethylene/kg fresh wt·hr is equivalent to an internal concentration of approximately 0.1 μl/l. Fumigation of fruiting cotton plants with only 0.5 μl/l caused 100% abscission of young fruits and floral buds within 2 days. This correlated with the estimated endogenous levels of ethylene. Reduced pressure, which reduced the internal levels of ethylene, delayed abscission of young fruits and leaves, a result which supports our conclusion from this study— that ethylene is one of the regulators of young fruit abscission in cotton.     

Abscission: the role of ethylene modification of auxin transport

The role of ethylene-mediated reduction of auxin transport in natural and ethylene-induced leaf abscission was studied in the cotton (Gossypium hirsutum L., cv. Stoneville 213) cotyledonary leaf system. The threshold level of ethylene required to cause abscission of intact leaves was between 0.08 and 1 μl/l with abscission generally occurring 12 to 24 hours following ethylene fumigation. The threshold level of ethylene required to reduce the auxin transport capacity in the cotyle-donary petiole paralleled that required for stimulation of abscission. In plants where cotyledons are allowed to senesce naturally there is a decline in auxin transport capacity of petioles and increase in ethylene synthesis of cotyledons. The visible senescence process which precedes abscission requires up to 11 days, and increases in ethylene production rates and internal levels were detected well before abscission. Ethylene production rates for entire cotyledons rose to 2.5 mμ1 g⁻¹ hr⁻¹ and internal levels of 0.7 μl/l were observed. These levels appear to be high enough to cause the observed decline in auxin transport capacity. These findings, along with those of others, indicate that ethylene has several roles in abscission control (e.g., transport modification, enzyme induction, enzyme secretion). The data indicate that ethylene modification of auxin transport participates in both natural abscission and abscission hastened by exogenous ethylene.     

Involvement of abscisic acid and ethylene in the responses of citrus seedlings to salt shock

The responses of salt‐sensitive citrus rootstocks to 200 m M NaCl were periodically determined on seedlings of citrange Carrizo ( Citrus sinensis [L.] Osbeck × Poncirus trifoliata [L.] Raf) during 30 days. The stressed seedlings adjusted osmotically, reduced stomatal conductance, increased proline content and ethylene production, and showed massive leaf abscission (92%). The salt shock also increased abscisic acid (ABA) and aminocyclopropane‐1‐carboxylic acid (ACC) in roots, xylem fluid and leaves, and in addition promoted Cl⁻ accumulation. The pattern of change of ABA, ACC and proline followed a two‐phase response: an initial transient increase (10‐12 days) overlapping with a gradual and continuous accumulation. This biphasic response appears to be compatible with the proposal that the transitory hormonal rises are induced by the osmotic component of salinity, whereas the Cl⁻ increase determines the subsequent accumulations. During the second phase, Cl⁻ levels correlated with abscission in leaves. Production of leaf ethylene was also concomitant with the increase in the abscission rate. Salt‐induced abscission was either reduced with CoCl₂ (52%) or inhibited with silver thiosulphate (14%). The results suggest that in salt‐stressed citrus, leaf abscission is induced by the chloride build‐up through a mechanism that stimulates leaf ACC synthesis and further conversion to ethylene.     

Fruit growth and abscission in Macadamia imtegrifolia

Growth in diameter of fruits (follicles) of five cultivate of Macadamia integrifolia Maiden and Betche followed a single sigmoidal pattern. A rapid increase in fruit diameter began 2 to 3 weeks after anithesis and continued until weeks 14 to 16. No further increase in size was observed after 16 weeks. Growth in fresh weight of the two cnltivars observed also followed a single sigmoidal pattern. A rapid increase in fresh weight began 5 to 6 weeks after anthesis and continued until weeks 15 to 18. No further increase in fresh weight was evident after 18 weeks. Abscission of flowers and fruits occurred continuously from anthesis through fruit maturity and could be separated into three periods. The first period of abscission occurred in the first 2 weeks after anthesis and commonly involved over 90% of the flowers. The second period of abscission occurred from 3 to 8 weeks after anthesis and commonly involved over 80% of the initial set of small fruit. This period of abscission occurred prior to and concurrent with the rapid increase in growth. The third period of abscission was gradual and occurred from 9 weeks after anthesis until maturity at 28 to 30 weeks. Measurement of ethylene evolution by fruits showed a peak during weeks 3 and 4. This was prior to the peak in relative abscission rate at week 5. The fruit removal force increased from anthesis through week 12 and decreased from week 24 through maturity.     

Source/Sink relations of abscising and nonabscising soybean flowers

The partitioning of recently fixed ¹⁴C to setting and abscising flowers within the axillary raceme of `Clark' isoline E1t soybeans (Glycine max L. Merr) was examined as a function of time after anthesis of individual flowers. In such racemes, the first four flowers showed a 17% abscission while the next four flowers showed 47% abscission.

Source/sink relations of flowers I-IV (normally setting) were compared to those of flowers V-VIII (normally abscising) by pulse labeling source leaves with ¹⁴CO₂ and determining the radioactivity of individual flowers after a 4-hour chase period. The relative specific activity (RSA;% disintegrations per minute per% dry weight), sink strength (% disintegrations per minute), and its components, sink size (milligrams dry weight) and sink intensity (% disintegrations per milligram dry weight) were then calculated as a function of days after anthesis.

Sink intensity (i.e. the competitive ability to accumulate photoassimilate per unit mass) was very high prior to anthesis of both setting and abscising flowers. Sink intensity then became very low for the first 3 days following anthesis after which it recovered in normally setting flowers, but failed to recover in normally abscising flowers. It is concluded that soybean reproductive abscission is determined at or very near the day of anthesis.