Exposure of Petunia Seedlings to Ethylene Decreased Apical Dominance by Reducing the Ratio of Auxin to Cytokinin

Seedlings of Petunia x hybrida ‘Orchid’ treated with the ethylene-releasing compound ethephon at 0.9, 1.7, and 3.5 mM evolved ethylene at a higher rate as the concentration of ethephon increased. Regardless of the concentration of ethephon applied, ethylene evolution peaked 6 to 8 h following application. Evidence that ethephon application decreased apical dominance included an increase in the number of new nodes on the main stem and a sustained increase in the length of new and existing lateral shoots compared to the control (no ethephon). Plants treated with 3.5 mM ethephon developed mild chlorosis, whereas a concentration of 1.7 mM ethephon decreased apical dominance without phytotoxic effects. The auxin/cytokinin ratio decreased in the apical shoot section as early as 1 h after ethephon treatment. In contrast, a decrease in the ratio in the subapical shoot section was not detected until 24 h after ethephon application. Reduction in auxin/cytokinin ratio was a result of a decrease in indole-3-acetic acid (IAA) and an increase of zeatin riboside (ZR), but not isopentenyladenosine (iPA). These results suggest that exposing ‘Orchid’ petunia seedlings to ethylene via ethephon lowers the auxin/cytokinin ratio, thereby promoting the outgrowth of lateral shoots.     

Influence of exogenous ethylene on cambial activity,xylogenesis and ray initiation in young shoots of Leucaena leucocephala (lam.) de Wit

The effect of exogenous ethephon on cambial activity, xylem production and ray population in young shoots of Leucaena leucocephala was investigated anatomically. The application of ethephon showed a diphasic effect on cambial activity and xylogenesis in a dose dependent manner. Lower concentration of ethephon enhanced cambial activity while high concentrations reduced cambial cell divisions and daughter-cell differentiation. High ethephon concentration also resulted in shorter vessel elements, thick walled fibers and phenolic accumulation in ray cells and vessel elements, whereas low concentration allowed elongation of fibers and vessel elements. The density of rays increased significantly with increase in ethylene concentration. The evaluation of longitudinal sections of cambial zone in ethephon treated plants showed high frequency of transformation of fusiform initials into ray initials through divisions and segmentation, resulting in high ray frequency in both xylem and phloem. The present study demonstrates that ethylene plays an important role in regulating secondary vascular tissue composition by reducing the population of fusiform initials in the cambium.     

Influence of ACC and Ethephon on cell growth in etiolated lupin hypocotyls. dependence on cell growth state

The possible implication of ethylene on the growth regulation of etiolated lupin hypocotyls was investigated. Excised hypocotyl sections from actively growing seedlings produced ethylene at a rate of 3 nmol h^-1 g^-1 min^-1. The rate of ethylene production was increased about 7 times when sections were treated with 10 mM 1-aminocyclopropane-1-carboxylic acid (ACC). Measurement of endogenous ACC showed that 95 % of total ACC (64.2 nmol g^-1 min^-1) corresponded to conjugated ACC. Treatments to intact seedlings with the ethylene precursor ACC, and the ethylene generating compound, 2-chloroethyl phosphonic acid (ethephon) during the cell elongation phase of the hypocotyl (from 7 to 21 dage), modified the cell growth of the organ. ACC (1 or 5 mM) or low concentrations of ethephon (0.66 mM) produced a transient decrease in the growth rate without modifying the final length of the hypocotyls. Higher concentrations of ethephon reduced the final length; the younger the seedlings were, the greater the reduction. Simultaneously to inhibition of cell elongation, ethephon produced stimulation of the radial expansion of cells in pith and cortex. The growth inhibition period, which lasted for 2 days after the treatments, was followed by another period in which the growth rate of treated plants surpassed that of the control. In both cases differences were observed along the hypocotyls due to the different growth status of the cells. It is suggested that the sensitivity to ethylene and the metabolism of ethylene depend on the growth status of the cells.     

Exposure of Petunia Seedlings to Ethylene Decreased Apical Dominance by Reducing the Ratio of Auxin to Cytokinin

Seedlings of Petunia x hybrida Orchid treated with the ethylene-releasing compound ethephon at 0.9, 1.7, and 3.5 mM evolved ethylene at a higher rate as the concentration of ethephon increased. Regardless of the concentration of ethephon applied, ethylene evolution peaked 6 to 8 h following application. Evidence that ethephon application decreased apical dominance included an increase in the number of new nodes on the main stem and a sustained increase in the length of new and existing lateral shoots compared to the control (no ethephon). Plants treated with 3.5 mM ethephon developed mild chlorosis, whereas a concentration of 1.7 mM ethephon decreased apical dominance without phytotoxic effects. The auxin/cytokinin ratio decreased in the apical shoot section as early as 1 h after ethephon treatment. In contrast, a decrease in the ratio in the subapical shoot section was not detected until 24 h after ethephon application. Reduction in auxin/cytokinin ratio was a result of a decrease in indole-3-acetic acid (IAA) and an increase of zeatin riboside (ZR), but not isopentenyladenosine (iPA). These results suggest that exposing Orchid petunia seedlings to ethylene via ethephon lowers the auxin/cytokinin ratio, thereby promoting the outgrowth of lateral shoots.     

Ethylene preparation and its application to physiological experiments

Ethylene is the first identified gaseous hormone regulating many aspects of plant growth and development. ACC and ethephon are two widely used chemicals replacing ethylene treatment when ethylene is not available. However, the amount of ethylene converted by ACC and ethephon is not controllable, leaving it questionable whether either treatment can mimic the effects of ethylene for experiments that are sensitive to ethylene concentration, response window, and treatment durations. Ethylene can be chemically made by ethanol dehydration; however, further purification from the dehydration products is needed. We previously reported that the ethylene gas can be easily prepared by decomposing ethephon in a buffered condition and the resulting ethylene can be used directly. Ethylene responses can be estimated by the measurement of the hypocotyl length of etiolated seedlings, or by ERF1 (Ethylene Response Factor1) expression. Although ACC of low concentrations is insufficient to induce ERF1 expression, ACC of high concentrations can replace ethylene for experiments where ethylene treatment is not feasible. However, ACC may undergo early consumption. Versatile approaches were developed so that laboratories lacking ethylene and techniques for gas handling can easily perform necessary ethylene treatments.Key words: ethylene preparation, ethephon, ACC     

Stimulation of Methionine Sulfoxide Reduction and Ethylene Production by Isoprothiolane in Relation to Its Activity against Non-parasitic Damping-off (MURENAE Disease) of Rice Seedlings

A rice blast controlling agent, isoprothiolane (diisopropyl 1,3-dithiolan-2-ylidenemalonate), stimulated the reduction of methionine sulfoxide to methionine by the rice plant. In the presence of isoprothiolane, the methionine/(methionine + its sulfoxide) ratio was increased to 129~208% of the control. The ethylene production by the plant was also enhanced by isoprothiolane, probably because methionine is an important precursor of ethylene. The non-parasitic damping-off caused by chilling stress on rice seedlings was effectively prevented with the application of isoprothiolane as well as ethephon, which easily decomposes to ethylene and acids. Therefore, the ethylene level modified by isoprothiolane and ethephon can contribute to their protective activity against the non-parasitic damping-off of rice seedlings. Indeed, a close relationship between the ethylene level and the protective activity against damping-off was obtained with isoprothiolane, but not with ethephon. Endogenous ethylene seems to be more effective in controlling the damping-off than exogenous ethylene from ethephon.     

A comparison of the effects of mechanically-induced stress,ethephon and silver thiosulphate on the growth of cauliflower seedlings

Mechanically-induced stress (MIS) was applied to cauliflower seedlings by brushing with paper for 1.5 minutes each day. With the possible exception of the effect on leaf 1 thickness, none of the growth responses induced by MIS were significantly nullified by spraying the seedlings with the ethylene inhibitor, silver thiosulphate (STS). The ethylene-releasing compound, ethephon, induced some changes in cauliflower growth similar to those caused by MIS, such as reduced shoot and leaf 1 weight, a reduction in cotyledon and leaf 1 area, and an increase in the thickness of leaves 1 and 2. However, other effects of ethephon were different from those of brushing. Petiole length and diameter and the weight and area of leaf 2 were reduced by brushing but generally increased by ethephon. STS reversed, at least partially, most of the ethephon effects on growth, thus demonstrating that it acts as an inhibitor of ethylene action in cauliflower.The results are discussed in relation to the possible endogenous control of MIS growth responses by endogenous ethylene and auxin.     

Ethylene is involved in the actin cytoskeleton rearrangement during the root gravitropic response of <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis>

Gravitropism, the directed plant growth with respect to the gravity vector, is regulated by auxin and its polar transport system, several secondary messengers, and by the cytoskeleton. Recently we have shown that the actin cytoskeleton in the root transition zone of Arabidopsis thaliana (L.) Heynh was rearranged after gravistimulation (rotation by 90°): the fraction of axially aligned microfilaments decreased and the fraction of oblique and transversally-oriented microfilaments increased. In the present research we have studied the effect of ethylene and inhibitors of its synthesis on actin cytoskeleton rearrangement during the gravitropic response. Application of the ethylene releasing substance ethephon to A. thaliana seedlings led to the disassembly of actin microfilaments as well as their broad angle distribution in cells of the root transition zone. This actin rearrangement was escaped by treatment with the ethylene synthesis inhibitor aminoethoxyvinylglycine (AVG). Another negative regulator of ethylene, salicylic acid, was shown to disturb actin microfilament rearrangement as well. We conclude that ethylene is essential for the process of actin cytoskeleton rearrangement in root cortex cells during the gravitropic bending response.     

Regulation of Expression of the prb-1b / ACC Deaminase Gene by UV-8 in Transgenic Tomatoes

Transgenic tomato plants with 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase gene from Enterobacter cloacae UW4 under the control of a pathogenesis-related promoter (prb-1b) from tobacco were challenged by abiotic stresses to determine the expression patterns ofthe transgene. No ACC deaminase RNA or protein was detected by RT-PCR and in western blots prepared from leaf proteins of transgenic plants after wounding or treatment with α-amino butyric acid, xylanase, ethephon, salicylic acid, jasmonic acid, ethylene, or ethylene plus jasmonic acid. However, expression of the ACC deaminase transgene was observed in leaves and roots oftransformed tomato lines exposed to UV light. The UV response required a minimum of 48 h of exposure and was specific to UV-8 light.     

Gravitropism in Higher Plant Shoots : IV. Further Studies on Participation of Ethylene

Ethylene at 1.0 and 10.0 cubic centimeters per cubic meter decreased the rate of gravitropic bending in stems of cocklebur (Xanthium strumarium L.) and tomato (Lycopersicon esculentum Mill), but 0.1 cubic centimeter per cubic meter ethylene had little effect. Treating cocklebur plants with 1.0 millimolar aminoethoxyvinylglycine (AVG) (ethylene synthesis inhibitor) delayed stem bending compared with controls, but adding 0.1 cubic centimeter per cubic meter ethylene in the surrounding atmosphere (or applying 0.1% ethephon solution) partially restored the rate of bending of AVG-treated plants. Ethylene increases in bending stems, and AVG inhibits this. Virtually all newly synthesized ethylene appeared in bottom halves of horizontal stems, where ethylene concentrations were as much as 100 times those in upright stems or in top halves of horizontal stems. This was especially true when horizontal stems were physically restrained from bending. Ethylene might promote cell elongation in bottom tissues of a horizontal stem or indicate other factors there (e.g. a large amount of `functioning' auxin). Or top and bottom tissues may become differentially sensitive to ethylene. Auxin applied to one side of a vertical stem caused extreme bending away from that side; gibberellic acid, kinetin, and abscisic acid were without effect. Acidic ethephon solutions applied to one side of young seedlings of cocklebur, tomato, sunflower (Helianthus annuus L.), and soybean (Glycine max [L.] Merr.) caused bending away from that side, but neutral ethephon solutions did not cause bending. Buffered or unbuffered acid (HCl) caused similar bending. Neutral ethephon solutions produced typical ethylene symptoms (i.e. epinasty, inhibition of stem elongation). HCl or acidic ethephon applied to the top of horizontal stems caused downward bending, but these substances applied to the bottom of such stems inhibited growth and upward bending—an unexpected result.     

1-Aminocyclopropane-1-Carboxylic Acid Transported from Roots to Shoots Promotes Leaf Abscission in Cleopatra Mandarin (Citrus reshni Hort. ex Tan.) Seedlings Rehydrated after Water Stress

The effect of water stress and subsequent rehydration on 1-aminocyclopropane-1-carboxylic acid (ACC) content, ACC synthase activity, ethylene production, and leaf abscission was studied in Cleopatra mandarin (Citrus reshni Hort. ex Tan.) seedlings. Leaf abscission occurred when drought-stressed plants were allowed to rehydrate, whereas no abscission was observed in plants under water stress conditions. In roots of water-stressed plants, a high ACC accumulation and an increase in ACC synthase activity were observed. Neither increase in ACC content nor significant ethylene production were detected in leaves of water-stressed plants. After rehydration, a sharp rise in ACC content and ethylene production was observed in leaves of water-stressed plants. Content of ACC in xylem fluid was 10-fold higher in plants rehydrated for 2 h after water stress than in nonstressed plants. Leaf abscission induced by rehydration after drought stress was inhibited when roots or shoots were treated before water stress with aminooxyacetic acid (AOA, inhibitor of ACC synthase) or cobalt ion (inhibitor of ethylene-forming enzyme), respectively. However, AOA treatments to shoots did not suppress leaf abscission. The data indicate that water stress promotes ACC synthesis in roots of Cleopatra mandarin seedlings. Rehydration of plants results in ACC transport to the shoots, where it is oxidized to ethylene. Subsequently, this ethylene induces leaf abscission.     

Role of aminocyclopropane-l-carboxylic acid in ethylene release by distal tissues following localized application of ethephon in Cucurbita pepo

The application of ethephon to a single leaf of Cucurbita pepo L. cv. Trailing Marrow plants caused a huge increase in ethylene production from the treated organ and an increased rate of ethylene production from other parts of the plant. These increases were particularly marked in the shoot apex and expanding leaf. Prior treatment with aminoethoxyvinylglycine (AVG), an ethylene biosynthesis inhibitor, blocked the increased production of ethylene at sites distant from the point of ethephon application. This strongly suggests that the increased ethylene production at these distant sites is due to ethylene biosynthesis and not a result of the translocation of ethylene released by the breakdown of ethephon at the site of application. Assays of 1-aminocyclopropane-l-carboxylic acid (ACC), an ethylene precursor, showed that it increased substantially after ethephon application but was at undetectable levels in the presence of AVG. It is proposed that the application of ethephon stimulates ethylene biosynthesis, but that transport through the plants is effected by ACC which is then converted to ethylene at the shoot apex and leaves.     

Polyamines in plants infected by citrus exocortis viroid or treated with silver ions and ethephon

The levels of polyamines in leaves of Gynura aurantiaca DC and tomato, Lycopersicon esculentum Mill. cv Rutgers, infected with citrus exocortis viroid (CEVd) or treated with silver nitrate or ethephon (2-chloroethylphosphonic acid) were measured by HPLC in relation to development of symptoms. Previously it had been demonstrated that treatment of G. aurantiaca plants with silver nitrate or ethephon closely mimicked the effects of viroid infection in the plants. In the studies reported here, a marked decrease in putrescine level was observed in plants infected by CEVd or treated with silver ions or ethephon. There was no significant change in either spermidine or spermine levels. Treatment of G. aurantiaca plants with specific inhibitors of ethylene biosynthesis (aminoethoxyvinylglycine, Co²⁺) or ethylene action (norbornadiene) prevented the decrease of putrescine associated with silver nitrate treatment and had no effect on spermidine or spermine levels. The development of viroid-like symptoms, the production of associated pathogenesis-related proteins, and the rise in protease activity induced by silver nitrate, were all suppressed by exogenous application of putrescine. The decreased level of putrescine as an ethylene-mediated step in the transduction of the viroid and silver or ethephon signaling is discussed.     

Phenolic acids and peroxidase activity in alfalfa (Medicago sativa) embryogenic cultures after ethephon treatment

Treatment with ethephon increased the concentration of exogenous ethylene in Medicago sativa L. embryogenic cell suspension cultures (consisting of single cells, small cellular clumps and globular somatic embryos) and induced changes in the metabolism of phenolic substances, activities of peroxidase (EC 1.11.1.7) and caused significant suppression of suspension culture growth. Treatment with the ethylene-releasing substance, ethephon, resulted in a several-fold increase in phenylalanine ammonia-lyase (PAL; EC 4.3.1.5) activity above the basal level and was accompanied by an elevated accumulation of phenolic acids (significant increase of methoxy-substituted acids). The majority of newly synthesised phenolic acids was incorporated into the fractions of glycosides and esters bound to the cell wall. Phenolic glycosides seemed to serve as a metabolic pool from which the phenolics were utilised during further culture. The increased activity of wall-bound ionic peroxidase after ethephon application correlated with the pronounced incorporation of ferulic acid in the cell walls. In contrast, the increased level of exogenous ethylene did not influence the growth of culture of more advanced embryos nor did it significantly alter phenylpropanoid metabolism.     

Ethylene perception generates gravicompetence in gravi-incompetent leaves of rye seedlings

The elongating leaves of young rye seedlings do not show a gravitropic response when placed horizontally. However, after treatment with ethylene, either supplied exogenously via ethephon or by application of its precursor 1-aminocyclopropane-1-carboxylic acid (ACC), gravicompetence is seen. The inhibition of ethylene perception by 1-methylcyclopropene (MCP) prevents gravicompetence. Young rye leaves provide a useful model system in which to identify the components of the gravity sensing or response systems, the presence of which govern gravicompetence.     

盐胁迫条件下提高内源乙烯对拟南芥幼苗生长和根尖活性氧水平的影响

以模式植物拟南芥（Arabidopsis thaliana）为材料,研究了内源乙烯对幼苗耐盐性的影响。研究结果表明,在施加了浓度为100 mmol·L^-1的NaCl胁迫的基质环境中,野生型拟南芥幼苗的根长和根重都显著减小。在施加外源乙烯利后不仅能够缓解盐胁迫对幼苗根伸长生长的抑制作用,而且能够缓解盐胁迫对幼苗根增重生长的抑制作用。施加外源ACC则只能缓解盐胁迫对幼苗根增重生长的抑制作用,而不能缓解盐胁迫对根的伸长生长的抑制。此外,100 mmol·L^-1 NaCl的胁迫条件下,拟南芥幼苗根尖中ROS水平明显升高,而施加了乙烯利和ACC处理下,幼苗根尖ROS的水平在NaCl胁迫下并没有明显的升高,说明内源乙烯可以调控植物体内的ROS维持在正常的水平,使植物体免受氧化损伤,从而提高了幼苗耐盐性。     

Influence of growth retardants on the internode and ethylene production of sunflower plants

The growth-retarding activity of the norbornenodiazetine tetcyclacis and the di-oxanylalkenyl triazole LAB 150 978 as well as the ethylene-forming compounds 2-chloroethyl-phosphonic acid (ethephon) and 1-amino-cyclopropane-l-carboxylic acid (ACC) on stem histogenesis and ethylene production of sunflower plants ( He-lianthus annuus L. cv.Spanners Allzweck) has been studied. The shoot growth of plants hydroponically grown and treated was reduced by the compounds. The shortening in the length of the 1st internode caused by tetcyclacis and LAB 150 978 was mainly induced by inhibition of cell division (the internode possessed fewer cortical cells per cell file). In contrast, ethephon and ACC decreased internode elongation mainly by reducing the rate of cell enlargement.
The ethylene production of sunflower seedlings cultivated on agar nutrient medium rose with increasing concentrations of ethephon and ACC, the shoot length of the plants being progressively reduced.
Tetcyclacis and LAB 150 978 inhibited both the formation of ethylene and shoot growth. It is suggested that in contrast to ethephon and ACC, tetcyclacis and LAB 150 978 do not achieve their growth-retarding effect by influencing the production of ethylene.     

Ethylene reverses photosynthetic inhibition by nickel and zinc in mustard through changes in PS II activity,photosynthetic nitrogen use efficiency,and antioxidant metabolism

We investigated the influence of exogenously sourced ethylene (200 μL L^?1 ethephon) in the protection of photosynthesis against 200 mg kg^?1 soil each of nickel (Ni)- and zinc (Zn)-accrued stress in mustard (Brassica juncea L.). Plants grown with Ni or Zn but without ethephon exhibited increased activity of 1-aminocyclopropane carboxylic acid synthase, and ethylene with increased oxidative stress measured as H₂O₂ content and lipid peroxidation compared with control plants. The oxidative stress in Ni-grown plants was higher than Zn-grown plants. Under metal stress, ethylene protected photosynthetic potential by efficient PS II activity and through increased activity of ribulose-1,5-bisphosphate carboxylase and photosynthetic nitrogen use efficiency (P-NUE). Application of 200 μL L^?1 ethephon to Ni- or Zn-grown plants significantly alleviated toxicity and reduced the oxidative stress to a greater extent together with the improved net photosynthesis due to induced activity of ascorbate peroxidase and glutathione (GSH) reductase, resulting in increased production of reduced GSH. Ethylene formation resulting from ethephon application alleviated Ni and Zn stress by reducing oxidative stress caused by stress ethylene production and maintained increased GSH pool. The involvement of ethylene in reversal of photosynthetic inhibition by Ni and Zn stress was related to the changes in PS II activity, P-NUE, and antioxidant capacity was confirmed using ethylene action inhibitor, norbornadiene.     

Uptake and Fate of Ethephon ([2-Chloroethyl]phosphonic Acid) in Dormant Weed Seeds

Although ethephon ([2-chloroethyl]phosphonic acid) is often used as a form of liquid ethylene in studies of seed germination, it is not known if ethylene evolved from ethephon in the seed is sufficient to elicit the desired response and/or if ethephon has a regulatory action that alone accounts for the response. For these reasons we studied the uptake and fate of [1,2-¹⁴C]ethephon in dormant seeds of Avena fatua, Sinapis arvensis, Thlaspi arvense, and Chenopodium album. The radioactivity within the seeds was separated into a labile carbon-labeled ethephon/ethylene fraction (64-87%) and, following extraction in methanol-chloroform-water (12:5:3), into fractions associated with insoluble (12-29%) and soluble (3-8%) seed constituents. The radioactivity associated with seed constituents was reduced 5 to 75% by hot alkaline hydrolysis (2.5 n KOH, 70° C for 1 hour). Although a small portion of the ethephon (or metabolite of ethephon/ethylene) taken up by the seeds is tightly bound to the tissues, our results indicate that, at the appropriate external concentrations of ethephon, the amount of ethylene evolved from ethephon within the seeds is sufficient to produce the desired ethylene mediated responses. However, factors affecting the decomposition of ethephon must be considered in the decision as to whether to use ethephon as a liquid supply of ethylene.