Abscission: the initial effect of ethylene is in the leaf blade

The leaf blade of cotton (Gossypium hirsutum L. cv. Stoneville 213) was investigated as the initial site of ethylene action in abscission. Ethylene applied at 14 μl/l to intact 3-week-old plants caused abscission of the third true leaf within 3 days. However, keeping only the leaf blade of this leaf in air during ethylene treatment of the rest of the plant completely prevented its abscission for up to 7 days. This inhibition of abscission was apparently the result of continued auxin production in the blade since (a) the application of an auxin transport inhibitor to the petiole of the air-treated leaf blade restored ethylene sensitivity to the leaf in terms of abscission; (b) repeated applications of naphthaleneacetic acid to the leaf blade of the third true leaf, when the entire plant was exposed to ethylene, had the same preventive effect on abscission of this leaf as keeping its leaf blade in air; and (c) the inhibitory effect of ethylene on auxin transport in the petiole, which is reduced by auxin treatment, was also reduced by placing the leaf blade in air.     

Starch Synthetase, Phosphorylase, ADPglucose Pyrophosphorylase, and UDPglucose Pyrophosphorylase in Developing Maize Kernels

Three types of whole plant experiments are presented to substantiate the concept that an important function of ethylene in abscission is to reduce the transport of auxin from the leaf to the abscission zone. (a) The inhibitory effect of ethylene on auxin transport, like ethylene-stimulated abscission, persists only as long as the gas is continuously present. Cotton (Gossypium hirsutum L. cv. Stoneville 213) and bean (Phaseolus vulgaris L. cv. Resistant Black Valentine) plants placed in 14 μl/l of ethylene for 24 or 48 hours showed an increase in leaf abscission and a reduced capacity to transport auxin; but when returned to air, auxin transport gradually increased and abscission ceased. (b) Ethylene-induced abscission and auxin transport inhibition show similar sensitivities to temperature. A 24-hour exposure of cotton plants to 14 μl/l of ethylene at 8 C resulted in no abscission and no significant inhibition of auxin transport. Increasing the temperature during ethylene treatment resulted in a progressively greater reduction in auxin transport with abscission occurring at [unk]27 C where auxin transport was inhibited over 70%. (c) Auxin pretreatment reduced both ethylene-induced abscission and auxin transport inhibition. No abscission occurred, and auxin transport was inhibited only 18% in cotton plants which were pretreated with 250 mg/l of naphthalene acetic acid and then placed in 14 μl/l of ethylene for 24 hours. In contrast, over 30% abscission occurred, and auxin transport was inhibited 58% in the corresponding control plants.     

Failure of Ethylene to Change the Distribution of Indoleacetic Acid in the Petiole of Coleus blumei X frederici during Epinasty

The effect of ethylene on the distribution of applied indoleacetic acid in the petiole of Coleus blumei Benth. X C. frederici G. Taylor has been investigated during the development of epinastic curvature. Using intact plants, ¹⁴C-IAA was applied to the distal region of the leaf lamina and the accumulation of label in the abaxial and adaxial halves of 5 mm petiole sections was determined after 1.5, 3, and 6 hours. Over this period the label was transported out of the lamina into the petiole at a rate of at least 66 mm hr⁻¹. Of the total amount of label in the petiole sections, 24 to 30% was located in the adaxial half and this distribution was not altered significantly by exposing plants to an atmosphere containing 50 μl/l ethylene. Thus when epinastic curvature is induced by ethylene there is no associated increase in the IAA content of the expanding adaxial half. The role of endogenous IAA in petiole epinasty was studied by restricting its movement with DPX 1840 (3,3a-dihydro-2-[p-methoxyphenyl]-8H-pyrozolo{5,1-a}isoindol-8-one). The leaf petioles still showed an initial epinastic response to ethylene. It is concluded that ethylene-induced epinasty is not dependent upon either any change in the transport of IAA or its redistribution within the petiole.     

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.     

Rapid growth inhibition of Avena coleoptile segments by abscisic Acid

An angular position sensing transducer was used to make continuous measurements of elongation of a column of Avena sativa coleoptile segments. Elongation stimulated by 2 μm indoleacetic acid was inhibited by 0.1 mm abscisic acid with a latent period of about 4 or 5 minutes at pH 6.0, 30 C. Full growth inhibition was not established until about 1 hour after the addition of the abscisic acid. The same degree of final growth inhibition could be obtained under the above conditions using 10 μM and 1 μM abscisic acid, but the latent period was longer. Pretreatments with abscisic acid affected the growth rate but did not extend the latent period of a subsequent response to auxin. The short term kinetics of inhibition by abscisic acid were not similar to those of any of the inhibitors of RNA and protein synthesis tested in this system.     

Functional Assessment of Cardiac Responses of Adult Zebrafish (Danio rerio) to Acute and Chronic Temperature Change Using High-Resolution Echocardiography

The zebrafish (Danio rerio) is an important organism as a model for understanding vertebrate cardiovascular development. However, little is known about adult ZF cardiac function and how contractile function changes to cope with fluctuations in ambient temperature. The goals of this study were to: 1) determine if high resolution echocardiography (HRE) in the presence of reduced cardiodepressant anesthetics could be used to accurately investigate the structural and functional properties of the ZF heart and 2) if the effect of ambient temperature changes both acutely and chronically could be determined non-invasively using HRE in vivo. Heart rate (HR) appears to be the critical factor in modifying cardiac output (CO) with ambient temperature fluctuation as it increases from 78 ± 5.9 bpm at 18°C to 162 ± 9.7 bpm at 28°C regardless of acclimation state (cold acclimated CA– 18°C; warm acclimated WA– 28°C). Stroke volume (SV) is highest when the ambient temperature matches the acclimation temperature, though this difference did not constitute a significant effect (CA 1.17 ± 0.15 μL at 18°C vs 1.06 ± 0.14 μl at 28°C; WA 1.10 ± 0.13 μL at 18°C vs 1.12 ± 0.12 μl at 28°C). The isovolumetric contraction time (IVCT) was significantly shorter in CA fish at 18°C. The CA group showed improved systolic function at 18°C in comparison to the WA group with significant increases in both ejection fraction and fractional shortening and decreases in IVCT. The decreased early peak (E) velocity and early peak velocity / atrial peak velocity (E/A) ratio in the CA group are likely associated with increased reliance on atrial contraction for ventricular filling.     

Enhancement of auxin sensitivity in Ranunculus sceleratus by ethylene: A mechanism to escape from hypoxia under temporary submergence

We analyzed auxin-induced and ethylene-enhanced elongation of petiole segments in Ranunculus sceleratus L. The early time course of elongation in petiolar segments was monitored with a computer-based video digitizer system. The application of ethylene-releasing ethrel slightly increased the elongation rate in the absence of IAA. When IAA alone was applied, elongation increased after a latent period of approximately 30 min. Maximal elongation rate was attained immediately after the latent period, and then the stabilized steady rate was recorded. During this phase, addition of ethrel strongly increased the elongation rate after a period of approximately 18 min. Although ethrel could acidify the growth medium, only a small part of the enhanced elongation was due to an acid-growth effect. Most of the growth stimulation was auxin-dependent and must be ascribed to the presence of ethylene. In the presence of ethrel, the log-concentration-response curve of IAA appeared to be shifted to the left. This kinetic analysis indicates an increase, due to ethylene, in the sensitivity of the R. sceleratus petiole to auxin, which results in inducing rapid growth to escape from hypoxia under temporary submergence.     

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.     

Free Methionine Levels in rin and Normal Isogenic Tomato Fruits Ripened in the Field or in Storage

Free methionine levels in rin and normal tomato fruits were determined microbiologically. Similar levels (1750 μg/100 g fresh weight) for mature green fruits of both rin and a normal isogenic line suggest that the lack of ripening of rin fruits is not due to low methionine levels. Methionine levels of mature green rin and normal fruits were 1750 μg/ 100 g fresh weight. Normal fruits ripened either on or off the vine were 2860 and 2500 μg/100 g fresh weight, respectively. The rin fruits which were left on the plant or held in air at 20 C until soft and yellow were significantly lower in methionine than C₂H₄-treated rin fruits or any normal fruits. Harvested rin and normal fruits held at 20 C in continuously applied ethylene (10 μl/l) had higher methionine levels than comparable air controls; levels in treated rin fruits were significantly higher than those in normal fruits.     

Autoinhibition of Ethylene Formation in Nonripening Stages of the Fruit of Sycomore Fig (Ficus sycomorus L.)

Differences in the mechanism of ethylene emanation of Ficus sycomorus L. during various stages of the fruit development were investigated by enclosing the figs in jars. Two distinct patterns of ethylene emanation were found. Pattern a. in stages not capable of ripening, neither spontaneously nor as a result of physiological treatment (nonripening stages A and C), ethylene concentration in the jar increased linearly for a short time and then remained constant. Pattern b. in stages capable of ripening (ripening stages B, D, and E), the linear increase in ethylene concentration continued for the entire period of measurement. In nonripening stages, ethylene emanation stopped when ethylene concentration in the jar reached a constant value (0.6 μl/l at stage C). Aeration of the figs and the jar renewed ethylene emanation. CO₂ concentration in the jar never exceeded 0.5%. Treatment of stage C figs with 0.6 to 10 μl/l exogenous ethylene caused immediate and complete cessation of ethylene emanation whereas the same treatment did not cause any change in rate of ethylene emanation from figs at the ripening stages B and D. Gashing (wounding) of stage C figs temporarily changed the pattern of ethylene emanation from pattern a to pattern b.     

Role of cytokinins in carnation flower senescence

Stem and leaf tissues of carnation (Dianthus caryophyllus) plants appear to contain a natural antisenescence factor since removal of most of these tissues from cut carnation flowers hastened their senescence. However, kinetin (5-10 μg/ml) significantly delayed senescence of flowers with stem and leaf tissues removed. In addition, the life span of cut flowers with intact (30-cm) stems was increased with kinetin treatment. Peak ethylene production by presenescent flowers was reduced 55% or more with kinetin treatment and was delayed by 1 day. Kinetin-treated flowers were less responsive to applied ethylene (100 μl/l for 3 hours) than untreated flowers. Possible natural roles of cytokinins in carnation flower senescence are discussed.     

Thermotolerance of Leaf Discs from Four Isoprene-Emitting Species Is Not Enhanced by Exposure to Exogenous Isoprene

The effects of exogenously supplied isoprene on chlorophyll fluorescence characteristics were examined in leaf discs of four isoprene-emitting plant species, kudzu (Pueraria lobata [Willd.] Ohwi.), velvet bean (Mucuna sp.), quaking aspen (Populus tremuloides Michx.), and pussy willow (Salix discolor Muhl). Isoprene, supplied to the leaves at either 18 μL L⁻¹ in compressed air or 21 μL L⁻¹ in N₂, had no effect on the temperature at which minimal fluorescence exhibited an upward inflection during controlled increases in leaf-disc temperature. During exposure to 1008 μmol photons m⁻² s⁻¹ in an N₂ atmosphere, 21 μL L⁻¹ isoprene had no effect on the thermally induced inflection of steady-state fluorescence. The maximum quantum efficiency of photosystem II photochemistry decreased sharply as leaf-disc temperature was increased; however, this decrease was unaffected by exposure of leaf discs to 21 μL L⁻¹ isoprene. Therefore, there were no discernible effects of isoprene on the occurrence of symptoms of high-temperature damage to thylakoid membranes. Our data do not support the hypothesis that isoprene enhances leaf thermotolerance.     

Rapid isolation of mesophyll cells from leaves of soybean for photosynthetic studies

Mesophyll cells were rapidly isolated from soybean (Glycine max [L.]) leaves using a combined Macerase enzyme-stirring technique. About 50% to 70% of the leaf cells on a chlorophyll basis from 3 grams of leaves could be isolated in 15 minutes. The cells obtained by this method were capable of high rates of photosynthesis even after storage in the dark for periods of up to 9 hours. The CO₂-saturated rate of photosynthesis increased from 5 μm CO₂/mg Chl·hour at 5 C to 170 μm CO₂/mg Chl·hour at 40 C. At atmospheric CO₂ concentration, the rate varied from 5 to 55 μm CO₂/mg Chl·hour over this temperature range. The reduced temperature response of photosynthesis at low CO₂ concentration was due to an increased Km(CO₂) of the cells with increasing temperature. The products of photosynthesis in the isolated cells were similar to the products of leaf photosynthesis.     

Rapidly Induced Wound Ethylene from Excised Segments of Etiolated Pisum sativum L., cv. Alaska: II. Oxygen and Temperature Dependency

Wound-induced ethylene synthesis by subapical stem sections of etiolated Pisum sativum L., cv. Alaska seedlings, as described by Saltveit and Dilley (Plant Physiol 1978 61: 447-450), was half-saturated at 3.6% (v/v) O₂ and saturated at about 10% O₂. Corresponding values for CO₂ production during the same period were 1.1% and 10% O₂, respectively. Anaerobiosis stopped all ethylene evolution and delayed the characteristic pattern of wound ethylene synthesis. Exposing tissue to 3.5% CO₂ in air in a flow-through system reduced wound ethylene synthesis by 30%. Enhancing gas diffusivity by reducing the total pressure to 130 mm Hg almost doubled the rate of wound ethylene synthesis and this effect was negated by exposure to 250 μl liter⁻¹ propylene. Applied ethylene or propylene stopped wound ethylene synthesis during the period of application, but unlike N₂, no lag period was observed upon flushing with air. It is concluded that the characteristic pattern of wound-induced ethylene synthesis resulted from negative feedback control by endogenous ethylene.

No wound ethylene was produced for 2 hours after excision at 10 or 38 C. Low temperatures prolonged the lag period, but did not prevent induction of the wound response, since tissue held for 2 hours at 10 C produced wound ethylene immediately when warmed to 30 C. In contrast, temperatures above 36 C prevented induction of wound ethylene synthesis, since tissue cooled to 30 C after 1 hour at 40 C required 2 hours before ethylene production returned to normal levels. The activation energy between 15 and 36 C was 12.1 mole kilocalories degree⁻¹.

     

Petiole Growth in Ranunculus sceleratus L.: Ethylene Synthesis and Submergence

Petioles of the celery-leaved buttercup (Ranunculus sceleratusL.) elongate in response to treatment with ethylene in air whenthe leaf blades are attached. An enhanced rate of elongationgrowth also occurs when the leaves are submerged. Submergencecauses an increase in extractable ethylene gas within the tissues,and these levels appear to approach those required to saturatethe ethylene-promoted elongation growth response. Coincidentwith a rise in ethylene in the tissues is a dramatic increasein the level of I-aminocyclopropane-1-carboxylic acid (ACC),the precursor of ethylene. Both the petiole and leaf blade tissueshave a similar capacity to evolve ethylene in the presence ofadded ACC. However, in air the leaf blade evolves more ethylenefrom endogenous resources than the petiole. The simultaneousincreases in ethylene and ACC levels in submerged tissues areconsidered in terms of the low diffusivity of ethylene in water,the ‘autocatalytic’ effect of ethylene on ethylenebiosynthesis and the rôle of both carbon dioxide and oxygenfluxes in ethylene metabolism of submerged tissues. Ranunculus sceleratus, celery-leaved buttercup, petiole growth, submergence, ethylene metabolism     

A kinetic analysis of hyponastic growth and petiole elongation upon ethylene exposure in Rumex palustris

Background and Aims

Complete submergence is an important stress factor for many terrestrial plants, and a limited number of species have evolved mechanisms to deal with these conditions. Rumex palustris is one such species and manages to outgrow the water, and thus restore contact with the atmosphere, through upward leaf growth (hyponasty) followed by strongly enhanced petiole elongation. These responses are initiated by the gaseous plant hormone ethylene, which accumulates inside plants due to physical entrapment. This study aimed to investigate the kinetics of ethylene-induced leaf hyponasty and petiole elongation.

Methods

Leaf hyponasty and petiole elongation was studied using a computerized digital camera set-up followed by image analyses. Linear variable displacement transducers were used for fine resolution monitoring and measurement of petiole growth rates.

Key Results

We show that submergence-induced hyponastic growth and petiole elongation in R. palustris can be mimicked by exposing plants to ethylene. The petiole elongation response to ethylene is shown to depend on the initial angle of the petiole. When petiole angles were artificially kept at 0°, rather than the natural angle of 35°, ethylene could not induce enhanced petiole elongation. This is very similar to submergence studies and confirms the idea that there are endogenous, angle-dependent signals that influence the petiole elongation response to ethylene.

Conclusions

Our data suggest that submergence and ethylene-induced hyponastic growth and enhanced petiole elongation responses in R. palustris are largely similar. However, there are some differences that may relate to the complexity of the submergence treatment as compared with an ethylene treatment.     

Elevated CO2 and Phosphate Limitation Favor Micromonas pusilla through Stimulated Growth and Reduced Viral Impact

Growth and viral infection of the marine picoeukaryote Micromonas pusilla was studied under a future-ocean scenario of elevated partial CO₂ (pCO₂; 750 μatm versus the present-day 370 μatm) and simultaneous limitation of phosphorus (P). Independent of the pCO₂ level, the ratios of M. pusilla cellular carbon (C) to nitrogen (N), C:P and N:P, increased with increasing P stress. Furthermore, in the P-limited chemostats at growth rates of 0.32 and 0.97 of the maximum growth rate (μ_max), the supply of elevated pCO₂ led to an additional rise in cellular C:N and C:P ratios, as well as a 1.4-fold increase in M. pusilla abundance. Viral lysis was not affected by pCO₂, but P limitation led to a 150% prolongation of the latent period (6 to 12 h) and an 80% reduction in viral burst sizes (63 viruses per cell) compared to P-replete conditions (4 to 8 h latent period and burst size of 320). Growth at 0.32 μ_max further prolonged the latent period by another 150% (12 to 18 h). Thus, enhanced P stress due to climate change-induced strengthened vertical stratification can be expected to lead to reduced and delayed virus production in picoeukaryotes. This effect is tempered, but likely not counteracted, by the increase in cell abundance under elevated pCO₂. Although the influence of potential P-limitation-relieving factors, such as the uptake of organic P and P utilization during infection, is unclear, our current results suggest that when P limitation prevails in future oceans, picoeukaryotes and grazing will be favored over larger-sized phytoplankton and viral lysis, with increased matter and nutrient flow to higher trophic levels.     

Upsurge in respiration and peroxide formation in potato tubers as influenced by ethylene, propylene, and cyanide

A continuous application of ethylene (10 μl/l) and propylene (500 μl/l) to potato tubers (Solanum tuberosum L.) resulted in an upsurge of respiration and a concomitant rise in peroxides. When applied in 100% O₂, the effect of ethylene and propylene on respiration and peroxide formation was augmented. Hydrogen cyanide (500 μl/l) mimicked the action of ethylene and propylene inducing a respiratory rise and a corresponding increase in peroxides. As with ethylene, the effect of HCN was augmented in high O₂ tensions. The results support the suggestion that ethylene activates the cyanide-insensitive respiratory pathway.     

Probing a Membrane Matrix Regulating Hormone Action: II. The Kinetics of Lipid-Induced Growth and Ethylene Production

Lipids which are active oleanimins, i.e., those which stimulate respiration and auxin-induced cell elongation of pea stem sections, also initiate a period of ethylene formation in them after a lag period of at least 1 hour. Production of ethylene requires auxin, is inhibited by cycloheximide and dinitrophenol applied during or before the lag period, and is greatly stimulated by lipids longer than 20 Ångstroms in length such as heptadecyl-benzene, chloro- or iodohexadecane, triolein, and vitamins E and K₁, but not by the shorter chloro- and iododecane. β-Stigmasterol at 10 to 40 μm concentrations depresses both oleanimin-induced growth and ethylene formation.     

Diurnal growth trends, water potential, and osmotic adjustment of maize and sorghum leaves in the field

The daily cycle of leaf elongation rate, water potential, and solute potential of maize and sorghum, as well as temperature, were monitored in the field. Major climatic features were high radiation and a minimum air temperature of about 12 C. Leaf elongation of both crops was slowest at night, presumably because of low temperature. Peak elongation rates were in daytime when leaf water potential (Ψ) was low. Solute potential also decreased during daylight, thus permitting the maintenance of appreciable turgor pressure, a critical parameter for cell expansion.