The role of ethylene in photoperiodic control of bulbing in Allium cepa

Bulb formation in Allium cepa L. cv. Dorata di Parma, was promoted by ethylene (Ethrel 120 mg l⁻¹) treatments. Application of the antiethylene complex silverthiosulphate (0.01 m M ) inhibited ethylene-induced bulb formation, but did not affect photoperiod-induced bulb formation. When the inhibitor of ethylene biosynthesis, aminoethoxyvinylglycine, was applied at 0.04 to 4 mg l⁻¹ to onion plants under inductive photoperiodic conditions the bulb promotion was unaffected. Therefore, Ethrel-induced bulb formation does not appear to be related to endogenous control of bulbing.     

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.     

Flower Abscission in Excised Inflorescences of Three Plectranthus Cultivars

Flower abscission induced by ethylene in three Plectranthus cultivars was investigated in order to characterise response to a range of inhibitory and antagonistic compounds. Excised inflorescences were exposed to 100 ml l⁻¹ ethylene gas or placed in various concentrations of ethephon (277, 27.7, 2.77, 0.277 and 0.0277 μM). Flower abscission in Plectranthus was readily induced by applying ethylene gas and by the 277 μM dose of ethephon. Removal of the inflorescences from the ethylene treatment prevented subsequent flower abscission. This implies that ethylene treatment did not induce an autocatalytic production of ethylene. Compounds that are known to compete for the ethylene receptor (100 and 500 ppb 1-methylcyclopropene or 100 and 500 ppm 2,5-norbornadiene) did not reduce abscission in this system. Also, application of the ethylene biosynthesis inhibitor, aminooxyacetic acid at 1 mM, was ineffective at preventing ethylene-induced flower abscission. In contrast, one compound known to block protein production (100 μM cycloheximide) and a non-competitive inhibitor of ethylene action (2 mM silver thiosulfate) did prevent ethylene-induced abscission. We conclude that flower abscission in cut inflorescences of Plectranthus is very likely mediated by endogenous ethylene production, but that control of ethylene-induced flower abscission in this genus can not be readily obtained by most ethylene antagonists that are known to be effective in other systems.     

Investigations on the role of ethylene in phytochrome-mediated photomorphogenesis

The etiolating, intact mustard (Sinapis alba L.) seedling exhibits a distinct temporal pattern of ethylene production. Light, operating through phytochrome, increases the rate of ethylene production without changing the pattern. Ethylene production of the isolated plant parts (segments), added together, exceed the production of the intact system even if the wound effect is taken into account. There is no significant light effect on ethylene production of the segments. Phytochrome-mediated anthocyanin synthesis in the cotyledons is inhibited by ethylene. The responsiveness towards ethylene of the anthocyanin producing metabolic chain is decreased by phytochrome. As anthocyanin synthesis is only partly inhibited under saturating ethylene concentrations in the atmosphere around the seedlings (100 l l^–1), a twofactor analysis becomes feasible. This analysis leads to the result that phytochrome and ethylene show multiplicative behavior, meaning that phytochrome and ethylene act on the same metabolic sequence (leading to anthocyanin) but independently of each other, and at different sites. Therefore, the hypothesis that ethylene mediates the action of phytochrome in anthocyanin synthesis and photomorphogenesis in general appears to be inapplicable.Abbreviations P_fr far-red absorbing form of phytochrome - P_r red absorbing form of phytochrome - P_tot total phytochrome, i.e. [P_r]+[P_fr]     

Immunolocalization of myosin in intact and wounded cells of the green algaErnodesmis verticillata (Kützing) Børgesen

Using in-vivo assays at least two classes of ethylene-binding site are shown to exist in pea epicotyls. These classes appear to have identical affinities for ethylene with a K_D of 6 · 10^–11-8 · 10^–11 M; the fast associating class has high and appropriate affinities for physiologically active analogues of ethylene, and CO₂ is without effect upon bfinding. Experiments involving suppression of endogenous ethylene production demonstrate that previous work may have underestimated both the amount of binding sites present and the affinity of such sites for ethylene. The apparent inhibition of binding by silver appears to be the consequence of a stimulation of endogenous ethylene production. The possible role of the sites as ethylene receptors is discussed.Abbreviations AOA amino oxyacetic acid - K_D dissociation constant We are grateful to the Biotechnology Action Programme of the European Economic Communities for support for this work.     

Studies on ethylene binding by cell free preparations from cotyledons of Phaseolus vulgaris L.

Various protein reactive agents such as dithioerythritol, dithiothreitol, mercaptoethanol and p-chloromercuribenzoate inhibit binding of ethylene to cell free preparations of Phaseolus vulgaris L. The effect of the thiols is partially reversed by treatment with diamide; occupation of the binding site by ligand diminishes the inhibition caused by p-chloromercuribenzoate but not that caused by thiols. Growth regulators other than ethylene do not affect binding. Physiologically active structural analogues of ethylene competitively inhibit binding of the growth regulator and their relative effectiveness in the cell free system closely resembles that in developmental processes controlled ethylene.Abbreviations DTE dithioerythritol - DTT dithiothreitol - EPPS N-2-hydroxyethyl-piperazine propane sulphonic acid - ME mercaptoethanol - PCMB p-chloromercuribenzoate     

Extended flower longevity of Petunia hybrida plants transformed with boers,a mutated ERS gene of Brassica oleracea

Petunia x hybridaHort.Vilm.-Andr. was transformed with boers, a mutatedallele of BOERS, an ethylene receptor sensor gene ofBrassica oleracea.boers was obtained by removing anEcoRI cutting site with a silent mutation at Gly-521 andintroducing a point mutation at Ile-62, replacing isoleucine withphenylalanine. Transformation was Agrobacterium tumefaciens mediated.Hygromycin resistant regenerants were tentatively confirmed as transformants byPCR's for HPH and boers and moredefinitively by Southern hybridization of genomic DNA with pBOERS4421. Flowersof transgenic plants retained turgidity and pigmentation considerably longerthan those of untransformed controls, whether left undisturbed on plants orexcised and placed in water. Furthermore, flowers were unaffected by exposureto exogenous ethylene. Excised shoots of transgenic plants released considerablymore ethylene than those of untransformed plants. Transformed plants alsoproduced apparently larger flowers. Unexpectedly higher mortality was observed,suggesting that the ethylene insensitive petunia plants were also lower indisease resistance.     

Ethylene and IAA interactions in the inhibition of photoperiodic flower induction of <Emphasis Type="Italic">Pharbitis nil</Emphasis>

It has been shown that both IAA and ethylene application inhibit flower induction in the short-day plant Pharbitis nil. However application of IAA has elevated ethylene production in this plant, as well. Strong enhancement of ethylene production is also correlated with the night-break effect, which completely inhibits flowering. In order to determine what the role of IAA and ethylene is in the photoperiodic flower induction in Pharbitis nil, we measured changes in their levels during inductive and non-inductive photoperiods, and the effects of ethylene biosynthesis and action inhibitors on inhibition of flowering by IAA. Our results have shown that the inhibitory effect of IAA on Pharbitis nil flowering is not physiological but is connected with its effect on ethylene biosynthesis.     

Ethylene and shoot regeneration: hookless1 modulates de novo shoot organogenesis in Arabidopsis thaliana

We have investigated the role of ethylene in shoot regeneration from cotyledon explants of Arabidopsis thaliana. We examined the ethylene sensitivity of five ecotypes representing both poor and prolific shoot regenerators and identified Dijon-G, a poor regenerator, as an ecotype with dramatically enhanced ethylene sensitivity. However, inhibiting ethylene action with silver nitrate generally reduced shoot organogenesis in ecotypes capable of regeneration. In ecotype Col-0, we found that ethylene-insensitive mutants (etr1-1, ein2-1, ein4, ein7) exhibited reduced shoot regeneration rates, whereas constitutive ethylene response mutants (ctr1-1, ctr1-12) increased the proportion of explants producing shoots. Our experiments with ethylene over-production mutants (eto1, eto2 and eto3) indicate that the ethylene biosynthesis inhibitor gene, ETO1, can act as an inhibitor of shoot regeneration. Pharmacological elevation of ethylene levels was also found to significantly increase the proportion of explants regenerating shoots. We determined that the hookless1 (hls1-1) mutant, a suppressor of the ethylene response phenotypes of ctr1 and eto1 mutants, is capable of dramatically enhancing shoot organogenesis. The effects of ACC and loss of HLS1 function on shoot organogenesis were found to be largely additive.     

Studies on the movement and distribution of ethylene in Vicia faba L.

In Vicia faba ethylene does not appear to move between different parts of the plant in physiologically significant amounts. The resistance to longitudinal movement is such that lateral emanation effectively isolates different parts of the plant from each other. When emanation is prevented, ethylene can be channelled to any part of the plant. Exposure of one section of a plant to ¹⁴C-labelled ethylene (up to 200 l/l) increased the internal concentration in other parts with ethylene that did not originate from the feeding chamber. A basipetal gradient of endogenous ethylene concentration was found in the lacuna of intact plants, the source of ethylene being the stem tissue. The permeability of stem tissue to ethylene decreases with age. The concentration of ethylene in tissues surrounding the lacuna is always higher than that in the lacuna and it is argued that compartmentation of ethylene occurs within these tissues.     

The effect of ammonium ions on uptake of glutamine and other amino compounds by cultured cells of rapeseed

Rooting responses and ethylene production by hypocotyl cuttings from etiolated mung-bean seedlings treated with the auxins -naphthaleneacetic acid, -(indole-3)-n-butyric acid (IBA) and 2,4,5-trichloro-phenoxypropionic acid were determined. There was no relationship between the abilities of the auxins to induce root formation and their capacities for inducing ethylene production. Studies with mixtures of 3-indoleacetic acid, a poor stimulator of rooting but an effective inducer of ethylene production, and IBA, an effective rooting stimulator but a poor inducer of ethylene production, exposure of cuttings to ethylene or (2-chloroethyl) phosphonic acid (Ethephon), hypobaric storage (150 mb) of treated cuttings, and exposure of auxin-treated cuttings to 7% CO₂ also indicated that ethylene is not directly involved in initiation of adventitious roots in this plant material.Abbreviations IAA indole-3-acetic acid - IBA -(indole-3)-n-butyric acid - NAA -naphthaleneacetic acid - 2,4,5-TP 2,4,5-trichlorophenoxypropionic acid     

Ethylene formation from 1-aminocyclopropane-1-carboxylic acid in homogenates of etiolated pea seedlings

Homogenates of etiolated pea (Pisum sativum L.) shoots formed ethylene upon incubation with 1-aminocyclopropane-1-carboxylic acid (ACC). In-vitro ethylene formation was not dependent upon prior treatment of the tissue with indole-3-acetic acid. When homogenates were passed through a Sephadex column, the excluded, high-molecular-weight fraction lost much of its ethylene-synthesizing capacity. This activity was largely restored when a heat-stable, low-molecular-weight factor, which was retarded on the Sephadex column, was added back to the high-molecular-weight fraction. The ethylene-synthesizing system appeared to be associated, at least in part, with the particulate fraction of the pea homogenate. Like ethylene synthesis in vivo, cell-free ethylene formation from ACC was oxygen dependent and inhibited by ethylenediamine tetraacetic acid, n-propyl gallate, cyanide, azide, CoCl₃, and incubation at 40°C. It was also inhibited by catalase. In-vitro ethylene synthesis could only be saturated at very high ACC concentrations, if at all. Ethylene production in pea homogenates, and perhaps also in intact tissue, may be the result of the action of an enzyme that needs a heat-stable cofactor and has a very low affinity for its substrate, ACC, or it may be the result of a chemical reaction between ACC and the product of an enzyme reaction. Homogenates of etiolated pea shoots also formed ethylene with 2-keto-4-mercaptomethyl butyrate (KMB) as substrate. However, the mechanism by which KMB is converted to ethylene appears to be different from that by which ACC is converted.Abbreviations ACC 1-aminocyclopropane-1-carboxylic acid - IAA indole-3-acetic acid - KMB 2-keto-4-mercaptomethyl butyrate - SAM S-adenosylmethionine     

Ethylene evolution by rust-infected,detached bean (Phaseolus vulgaris L.) leaves susceptible and hypersensitive to Uromyces phaseoli (Pers.) Wint.

Ethylene production in leaves of susceptible and hypersensitive varieties of beans has been followed after inoculation with Uromyces phaseoli. Four different states of ethylene evolution are distinguishable: (1) 13 h after inoculation and concomitant to the penetration of the fungal mycelium through the stomata, all varieties show an outburst of ethylene with significant differences between the three varieties. (2) After 36 h postinoculation, in all three varieties ethylene evolution is scarcely higher than in noninfected leaves. (3) Starting 59 h after inoculation, only in the hypersensitive variety 765 (which shows the lowest ethylene production after 13 h), a second, very strong ethylene outburst is observed. (4) From 125 h after inoculation, significant ethylene production is not observed in any variety. At this time, characteristic symptoms are expressed in susceptible leaves (differentiation of uredosori) and in the hypersensitive variety 765 (large brown necrotic spots); no macroscopic symptoms are observed in the hypersensitive variety 814, which exhibits the strongest ethylene outburst 13 h after inoculation. The capacity for ethylene formation after mechanical wounding (point freezing) is almost identical in healthy leaves of all three varieties. This capacity is still preserved after the first ethylene outburst 36 h after infection.This work was supported by the Deutsche Forschungsgemeinschaft, by the Kleinwanzlebener Saatzucht AG (Einbeck, FRG) and by a NATO fellowship to P.M.     

乙烯调控植物耐盐性的研究进展

乙烯具有复杂的生物学功能,它调节着植物生长发育和许多的生理生化过程。乙烯也被认为是一种胁迫应答激素,直到近几年关于乙烯生物合成及信号转导途径与植物盐胁迫的关系才逐渐被挖掘出来。乙烯在不同水平、层次参与盐胁迫反应,包括乙烯合成关键酶(ACS)和乙烯受体,细胞质中CTR1和EIN2以及细胞核中EIN3传导、响应盐信号。但是乙烯合成和信号转导途径在植物盐胁迫响应过程中仍然存在许多未解决的问题。主要介绍乙烯合成及信号转导途径的各组分与盐胁迫关系的最新研究进展,并讨论其存在的主要问题。     

D-Amino-acid-stimulated ethylene production in seed tissues

Ethylene production by axial and cotyledonary tissues excised from Xanthium pennsylvanicum Wallr. seeds was markedly (up to 5-fold) stimulated by the D-isomers of phenylalanine, valine, leucine, threonine, methionine and eithionine while the L-isomers caused no such effect. Responsiveness of these seed tissues to D-methionine appeared soon after the beginning of imbibition, reached a maximum after 6–12 and 12–24 h for the axial and cotyledonary tissues, respectively, and then decreased sharply. D-Phenylalanine and D-methionine also stimulated ethylene production in seed tissues of X. canadense Mill. and in cotyledonary segments from seeds of Helianthus annuus L., Cucurbita moschata Duch. and Vigna radiata (L.) Wilczek. The endogeneous ethylene production and the D-amino-acid-stimulated ethylene production by the seed segments was strongly inhibited by aminoethoxyvinyl glycine, a potent inhibitor of ethylene synthesis from L-methionine.     

Rapidly induced ethylene formation after wounding is controlled by the regulation of 1-aminocyclopropane-1-carboxylic acid synthesis

Bean leaves from Phaseolus vulgaris L. var. Pinto 111 react to mechanical wounding with the formation of ethylene. The substrate for wound ethylene is 1-aminocyclopropane-1-carboxylic acid (ACC). It is not set free by decompartmentation but is newly synthesized. ACC synthesis starts 8 to 10 min after wounding at 28°C, and 15 to 20 min after wounding at 20°C. Aminoethoxyvinylglycine (AVG), a potent inhibitor of ethylene formation from methionine via ACC, inhibits wound ethylene synthesis by about 95% when applied directly after wounding (incubations at 20°C). AVG also inhibits the accumulation of ACC in wounded tissue. AVG does not inhibit conversion of ACC to ethylene. Wound ethylene production is also inhibited by cycloheximide, n-propyl gallate, and ethylenediaminetetraacetic acid.Abbreviations ACC 1-aminocyclopropane-1-carboxylic acid - AVG ammoethoxyvinylglycine - EDTA ethylenediaminetetraacetic acid     

An increase in ethylene sensitivity is associated with jasmonate-promoted senescence of detached rice leaves

The role of ethylene in jasmonate-promoted senescence of detached rice leaves was investigated. Ethylene production in methyl jasmonate-treated leaf segments of rice was lower than in the control leaves. Treatment of leaf segments with silver nitrate or/and silver thiosulfate, inhibitors of ethylene action, inhibited methyl jasmonate-, jasmonic acid-, linolenic acid-, and abscisic acid-promoted senescence of detached leaves. We suggest that an increase in ethylene sensitivity, but not ethylene level, is the initial event triggering the enhanced senescence by jasmonates of detached rice leaves.Abbreviations JA jasmonic acid - MJ methyl jasmonate - STS silver thiosulfate - ABA abscisic acid     

The compartmentation of ethylene in developing cotyledons of Phaseolus vulgaris L.

Isolated cotyledons of Phaseolus vulgaris L. cv. Canadian Wonder accumulated ¹⁴C₂H₄ (0.7–1 l l^-1) from air to give partition coefficients of 1 to 4, which greatly exceeded the value obtained with steam killed cotyledons (0.05) and with water (0.11). After ¹⁴C₂H₄ treatment, 98% of the ¹⁴C in the tissue remained as ¹⁴C₂H₄. The labelled ethylene accumulated by cotyledons was released only slowly (1–10% h^-1) either in an air stream or into toluene. Heating to 60°C for 2 h, but not freezing and thawing, caused the immediate release of ¹⁴C₂H₄ from the tissue. Propylene and vinyl chloride competitively inhibited the accumulation of ¹⁴C₂H₄.Cotyledons emanated endogenous ethylene at a very low rate but after heating (although not freezing and thawing) 13 nl of ethylene per g fresh mass were released within minutes. It was concluded that french bean cotyledons hold ethylene in a compartmented form in sufficient amount to account for at least 200 h of emanation.Abbreviation PPO diphenyloxazole