Ethylene as an effector of wound-induced resistance to cellulase in oat leaves

Peeling the abaxial epidermis from oat leaves (Avena sativa var. Victory) induces the formation of wound ethylene and the development of resistance to cellulolytic digestion of mesophyll cell walls. Ethylene release begins between 1 and 2 hours after peeling in the light or dark. Aminoethoxyvinylglycine (AVG, 0.1 millimolar), CoCl₂ (1.0 millimolar), propyl gallate (PG, 1.0 millimolar) or aminooxyacetic acid (AOA, 1.0 millimolar) inhibits, whereas AgNO₃ stimulates wound ethylene formation. Incubation on inhibitors of ethylene biosynthesis (AVG, CoCl₂, PG, AOA) or action (AgNO₃, hypobaric pressure or the trapping of ethylene with HgClO₄) also prevents the development of wound-induced resistance to enzymic cell wall digestion. 1-Aminocyclopropane-1-carboxylic acid (ACC, 1.0 millimolar) reverses AVG (0.1 millimolar) inhibition of the development of resistance. Exogenous ethylene partially induces the development of resistance in unwounded oat leaves.

These results suggest that peeling of oat leaves induces ethylene biosynthesis, which in turn effects changes in the mesophyll cells resulting in the development of resistance to cellulolytic digestion.

     

Ethylene inhibitors enhance in vitro root formation from apple shoot cultures

Effects of the ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC) and three ethylene inhibitors, AgNO₃, aminoethoxyvinyglycine (AVG) and CoCl₂, on root formation were tested in vitro using shoot cultures of the apple (Malus×domestica Borkh.) cultivar Royal Gala. ACC inhibited root formation by delaying root emergence and increasing callus formation at the bases of shoots. In contrast, ethylene inhibitors promoted root formation. Both AgNO₃ and AVG at the appropriate concentrations increased the percentage of shoots producing roots and reduced callus formation at the base of these shoots. AgNO₃ stimulated root emergence and enhanced root growth, while AVG increased the number of roots per shoot. CoCl₂ slightly increased root number and rooting efficiency. These promotive effects may result from a reduction in ethylene concentration or inhibition of ethylene action. The results found in this study may be used to improve the rooting efficiency of other apple cultivars and rootstocks, and possibly of other plant species. Received: 2 March 1997 / Revision received: 1 July 1997 / Accepted: 18 July 1997     

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     

Effect of the defoliant thidiazuron on ethylene evolution from mung bean hypocotyl segments

The effect of the defoliant thidiazuron (N-phenyl-N′1,2,3-thiadiazol-5-ylurea) on ethylene evolution from etiolated mung bean hypocotyl segments was examined. Treatment of hypocotyl segments with concentrations of thidiazuron equal to or greater than 30 nanomolar stimulated ethylene evolution. Increased rates of ethylene evolution from thidiazuron-treated tissues could be detected within 90 minutes of treatment and persisted up to 30 hours after treatment. Radioactive methionine was readily taken up by thidiazuron-treated tissues and was converted to ethylene, 1-aminocyclopropane-1-carboxylic acid (ACC) and an acidic conjugate of ACC. Aminoethoxyvinylglycine, aminooxyacetic acid, cobalt chloride, and α-aminoisobutyric acid reduced ethylene evolution from treated tissues. An increase in the endogenous content of free ACC coincided with the increase in ethylene evolution following thidiazuron treatment. Uptake and conversion of exogenous ACC to ethylene were not affected by thidiazuron treatment. No increases in the extractable activities of ACC synthase were detected following thidiazuron treatment.     

Ethylene Evolution Changes in Tilted Fraxinus mandshurica Rupr. var. japonica Maxim. Seedlings in Relation to Tension Wood Formation

The effects of ethylene on tension wood formation were studied in 3-year-old Fraxinus mandshurica Rupr. var. japonica Maxim, seedlings in two separate experiments. In experiment 1, ethylene evolution of buds and stems was measured using gas chromatography after 0, 2,4, 7,14, and 21 d of treatment; in experiment 2, both aminoethoxyvinylglycine (AVG) and AgNO3 were applied to the horizontally-placed stems, and the cell numbers on sites of applications were measured after 40 d. Ethylene evolution from buds was found to be much greater in tilted seedlings than in upright ones. The cell numbers of wood fibers in shoots and 1-year-old stems were reduced in treatments with 12.5 x 10-7 μmol/L AVG, 12.5 x 10-8 μmol/L AVG, and 11.8 x 10-8 μmol/L AgNO3; whereas the horizontal and vertical diameters were reduced by treatment of 12.5 x 10-7 μnol/L AVG. Ethylene evolutions of shoots and 1-year-old stems were inhibited greatly in comparison with the control by applying 12.5 x 10-7 μmol/L AVG. The formation of a gelatinous layer of wood fibers was affected by neither AVG nor AgNO3 application. These results suggest that ethylene regulates the quantity of wood production, but does not affect G-layer formation in F. mandshurica Rupr. var. japonica Maxim, seedlings.     

Regulation of Auxin-induced Ethylene Biosynthesis. Repression of Inductive Formation of 1-Aminocyclopropane-1-carboxylate Synthase by Ethylene

Changes in the 1-aminocyclopropane-1-carboxylate (ACC) synthaseactivity which regulates auxin-induced ethylene production werestudied in etiolated mung bean hypocotyl segments. Increasesboth in ethylene production and ACC synthase activity in tissuetreated with IAA and BA were severely inhibited by cycloheximide(CHI), 2-(4-methyl-2,6-dinitroanilino)-N-methylpropionamide,actinomycin D and -amanitin. Aminoethoxyvinylglycine (AVG),a potent inhibitor of the ACC synthase reaction, increased theactivity of the enzyme in the tissue 3- to 4-fold. This stimulationalso was severely inhibited by the above inhibitors. Stimulationof the increase in the enzyme content by AVG was partially suppressedby an exogenous supply of ACC or ethylene. Suppression of theincrease in the enzyme took place with 0.3 µl/liter ethylene,and inhibition was increased to 10 µl/liter, which caused65% suppression. Air-flow incubation of the AVG-treated tissue,which greatly decreased the ethylene concentration surroundingthe tissue, further increased the amount of enzyme. Thus, oneeffect of AVG is to decrease the ethylene concentration insidethe tissue. The apparent half life of ACC synthase activity,measured by the administration of CHI, was estimated as about25 min. AVG lengthened the half life of the activity about 2-fold.Feedback repression by ethylene in the biosynthetic pathwayof auxin-induced ethylene is discussed in relation to the effectof AVG. (Received January 22, 1982; Accepted March 26, 1982)     

Ethylene release from green spruce needles involves a combination of ACC-dependent and independent pathways

Aminoethoxyvinylglycine (AVG) and cobalt ions strongly inhibit the conversion of added methionine or aminocyclopropane-1-carboxylic acid (ACC) into ethylene by green-coloured, non-stressed Norway spruce (Picea abies L.) needles but only 30%–40% of basal ethylene formation is affected by such inhibitors. In addition, free radical-mediated ACC-independent ethylene formation (AIEF) of the type released by brown-coloured spruce needles also occurs in extracts from healthy green-coloured needles. Treatment with CdCl₂ (10 mM), Na₂S₂O₅ (5 mM) or FeSO₄ (10 mM) induces 3–7 fold increases in the rates of ethylene evolution from green-coloured needles. However, only Cd²⁺-induced ethylene formation is inhibited by AVG while ethylene induced by S₂O₅ ^2- or Fe²⁺ is insensitive to added AVG although increased levels of ACC have also been detected in these treatments. Nevertheless, ethylene-forming decomposition of the precursors of AIEF is accelerated by S₂O₅ ^- or Fe²⁺ which indicates that the ethylene released from green-coloured spruce needles is formed by a combination of both the ACC-dependent and AIEF pathways.Abbreviations ACC 1-aminocyclopropane-1-carboxylic acid - AVG aminoethoxyvinylglycine - AIEF ACC-independent ethylene formation - EFE ethylene-forming enzyme - MACC N-malonyl(amino)cyclopropane-1-carboxylic acid - DTBN di-tert-butylnitroxide - MNP 2-methyl-2-nitrosopropane - SAM S-adenosylmethionine - TEMPO 2,2,6,6-tetramethyl-1-piperidine-N-oxyl     

Ethylene evolution from tobacco leaves irradiated with UV-B

Seedlings of Nicotiana tabacum L. (cv. Petit Havana SR1) were grown in the presence or absence of ultraviolet-B (UV-B, 290–320 nm) irradiation. The evolution of ethylene from the leaves, the content of 1-aminocyclopropane-1-carboxylic acid (ACC), an endogenous precursor of ethylene, and the activity of ACC synthase, a rate-limiting step in the production of ethylene, were increased by UV-B irradiation. The time course of these increases was parallel with the emergence of damage that was estimated by measuring the chlorophyll (Chl) content and the leakage of ions from leaf cells. Treatment of leaves with aminoethoxy-vinyl-glycine (AVG), a specific inhibitor of ACC synthase, reduced the extent of damage caused by UV-B. These results suggest that ethylene acts on certain processes to cause damage in tobacco leaves irradiated with UV-B. Electronic Publication     

Wound-induced Ethylene Formation in Albedo Tissue of Citrus Fruit

Excised albedo tissue of citrus fruit (Citrus unshiu and Citrus hassaku) produced ethylene at an increasing rate in response to wounding and aging. The application of 1-aminocyclopropane-1-carboxylic acid (ACC) enhanced ethylene production in both the fresh and aged tissues, but this increase was greater in the aged tissue than in the fresh tissue. ACC content was very low in fresh tissue but increased greatly in aging tissue, paralleling the rise in ethylene production. Aminoethoxyvinylglycine (AVG) strongly inhibited ethylene production in the aged tissue. In the presence of ACC, however, ethylene production was not inhibited by AVG. These results suggest that ACC is an intermediate in the pathway of ethylene biosynthesis in the albedo tissue and that both steps of ACC formation and ACC conversion to ethylene are enhanced by wounding and aging. Inhibitors of protein synthesis, cycloheximide and 2-(4-methyl-2,6-dinitroanilino)-N-methyl propionamide, strongly inhibited ethylene production in the albedo tissue, implying that protein synthesis is required to maintain the continuous evolution of ethylene. The stimulation of ethylene production by ACC was reduced by the addition of l-methionine, whereas d-methionine had very little inhibitory effect. Ethylene production in the albedo tissue was also inhibited by the addition of n-propyl gallate and 3,5-dibromo-4-hydroxybenzoic acid.     

Effect of root zone carbon dioxide enrichment on ethylene inhibition of carbon assimilation in potato plants

Potato plants ( Solanum tuberosum L. var. Russet Burbank) treated with 1 μl ethylene 1⁻¹ of air showed an inhibition of CO₂ assimilation by 18%. The inhibition occurred after 3 h of exposure to ethylene and was not mediated through closure of the stomata. The enrichment of the root zone with CO₂ almost completely abolished the ethylene inhibition of CO₂ assimilation which was apparently due to an increase in the intercellular concentration of CO₂ in leaves following enrichment. The effect of application of CO₂ to the root zone on ethylene inhibition of CO₂ assimilation seemed to last for a few days. Potato plants treated with aminoethoxyvinlglycine (AVG) showed an increase in fresh and dry weight as compared to non-treated plants. Our results indicate that both CO₂ and AVG alter the effect of ethylene and promote growth in plants by inhibiting ethylene action and biosynthesis, respectively.     

Role of Ethylene on de Novo Shoot Regeneration from Cotyledonary Explants of Brassica campestris ssp. pekinensis (Lour) Olsson in Vitro

The promotive effect of AgNO₃ and aminoethoxyvinylglycine (AVG) on in vitro shoot regeneration from cotyledons of Brassica campestris ssp. pekinensis in relation to endogenous 1-amino-cyclopropane-1-carboxylic acid (ACC) synthase, ACC, and ethylene production was investigated. AgNO₃ enhanced ACC synthase activity and ACC accumulation, which reached a maximum after 3 to 7 days of culture. ACC accumulation was concomitant with increased emanation of ethylene which peaked after 14 days. In contrast, AVG was inhibitory to endogenous ACC synthase activity and reduced ACC and ethylene production. The promotive effect of AVG on shoot regeneration was reversed by 2-chloroethylphosphonic acid at 50 micromolar or higher concentrations, whereas explants grown on AgNO₃ medium were less affected by 2-chloroethylphosphonic acid. The distinctive effect of AgNO₃ and AVG on endogenous ACC synthase, ACC, and ethylene production and its possible mechanisms are discussed.     

Involvement of ethylene in the action of the cotton defoliant thidiazuron

The effect of the defoliant thidiazuron (N-phenyl-N′-1,2,3-thiadiazol-5-ylurea) on endogenous ethylene evolution and the role of endogenous ethylene in thidiazuron-mediated leaf abscission were examined in cotton (Gossypium hirsutum L. cv Stoneville 519) seedlings. Treatment of 20- to 30-day-old seedlings with thidiazuron at concentrations equal to or greater than 10 micromolar resulted in leaf abscission. At a treatment concentration of 100 micromolar, nearly total abscission of the youngest leaves was observed. Following treatment, abscission of the younger leaves commenced within 48 hours and was complete by 120 hours. A large increase in ethylene evolution from leaf blades and abscission zone explants was readily detectable within 24 hours of treatment and persisted until leaf fall. Ethylene evolution from treated leaf blades was greatest 1 day posttreatment and reached levels in excess of 600 nanoliters per gram fresh weight per hour (26.7 nanomoles per gram fresh weight per hour). The increase in ethylene evolution occurred in the absence of increased ethane evolution, altered leaf water potential, or decreased chlorophyll levels. Treatment of seedlings with inhibitors of ethylene action (silver thiosulfate, hypobaric pressure) or ethylene synthesis (aminoethoxyvinylglycine) resulted in an inhibition of thidiazuron-induced defoliation. Application of exogenous ethylene or 1-aminocyclopropane-1-carboxylic acid largely restored the thidiazuron response. The results indicate that thidiazuron-induced leaf abscission is mediated, at least in part, by an increase in endogenous ethylene evolution. However, alterations of other phytohormone systems thought to be involved in regulating leaf abscission are not excluded by these studies.     

Ethylene formation in Pisum sativum and Vicia faba protoplasts

Protoplasts isolated from leaves of peas (Pisum sativum L.) and of Vicia faba L. produced 1-aminocyclopropane-1-carboxylic acid (ACC) from endogenous substrate. Synthesis of ACC and conversion of ACC to ethylene was promoted by light and inhibited by 3-(3,4-dichlorophenyl)-1,1-dimethylurea and carbonyl cyanide m-chlorophenylhydrazone. Aminoethoxyvinylglycine inhibited ethylene synthesis to a minor extent when given during incubation of the protoplasts but was very effective when added both to the medium in which the protoplasts were isolated and to the incubation medium as well. Radioactivity from [U-¹⁴C]methionine was incorporated into ACC and ethylene. However, the specific radioactivity of the C-2 and C-3 atoms of ACC, from which ethylene is formed, increased much faster than the specific radioactivity of ethylene. It appears that ACC and ethylene are synthesized in different compartments of the cell and that protoplasts constitute a suitable system to study this compartmentation.Abbreviations ACC 1-Aminocyclopropane-1-carboxylic acid - AVG aminoethoxyvinylglycine - CCCP carbonyl cyanide m-chlorophenylhydrazone - DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea     

Ethylene evolution from various developing organs of olive (Olea europaea) after excision

Ethylene evolution from leaves, stems, inflorescences and fruits of the olive plant ( Olea europaea L.) cv. Manzanillo was studied at various stages of their development. Mature non-growing organs, particularly leaves, have a constant, low, and uniform rate of ethylene evolution. Ethylene evolution from detached mature olive leaves was constant during the first 12 h after excision. Leaves on shoots maintained in vitro kept a constant rate of ethylene evolution for at least the first 5–6 days. Leaf injury significantly increased ethylene evolution. Ethylene evolution from injured and non-injured control leaves could be markedly inhibited aminoethoxyvinylglycine (AVG) applied to the leaves or fed to the shoot. The use of excised olive shoots and leaves as an in vitro model system for studies of induced metabolic processes such as abscission and developing water stress was suggested.     

Atmospheric Ozone Interacts with Stress Ethylene Formation by Plants to Cause Visible Plant Injury

Ozone toxicity was studied in peas, beans, and tobacco (BelB and Bel W3). These experiments showed that ozone toxicitywas related to the rates of ethylene biosynthesis. Sensitivityto ozone was reduced if ethylene biosynthesis was inhibitedafter treatment with aminoethoxyvinylglycine (AVG). Similarly,plants that were able to reduce or prevent stress ethylene formationwere less sensitive after both short- and long-term exposureto ozone. Plants conditioned by longer exposures to ozone havelow rates of ethylene formation and this may be why brief ozoneexposures may be more phytotoxic than prolonged fumigations. Key words: Nicotiana tabacum, Phaseolus vulgaris, Pisum sativum, lipid peroxidation, peas, Pinto beans, tobacco     

Ethylene Promotes Elongation Growth and Auxin Promotes Radial Growth in Ranunculus sceleratus Petioles

Submergence induces elongation in the petioles of Ranunculus sceleratus L., after a rise in endogenous ethylene levels in the tissue. Petioles of isolated leaves also elongate 100% in 24 hours when treated with ethylene gas, without a change in the radius. Application of silver thiosulfate, aminoethoxyvinylglycine (AVG), abscisic acid (ABA), or methyl jasmonate inhibits this elongation response. Gibberellic acid treatment promotes ethylene-induced elongation, without an effect on the radius. Indoelastic acid (IAA) induces radial growth in the petioles, irrespective of the presence or absence of added ethylene. High concentrations of IAA will also induce elongation growth, but this is largely due to auxin-induced ethylene synthesis; treatment with silver thiosulfate, AVG, ABA, or methyl jasmonate inhibit this auxin-promoted elongation growth. However, the radial growth induced by IAA is not affected by gibberellic acid, and not specifically inhibited by ABA, methyl jasmonate, silver thiosulfate, or AVG. These results support the idea that petiole cell elongation during “accommodation growth” can be separated from radial expansion. The radial expansion may well be regulated by IAA. However, effects of high levels of IAA are probably anomalous, since they do not mimic normal developmental patterns.     

Disruption of the Polar Auxin Transport System in Cotton Seedlings following Treatment with the Defoliant Thidiazuron

The effect of the defoliant thidiazuron (TDZ) on basipetal auxin transport in petiole segments isolated from cotton (Gossypium hirsutum L. cv LG102) seedlings was examined using the donor/receiver agar block technique. Treatment of intact seedlings with TDZ at concentrations of 1 micromolar or greater resulted in a dose-dependent inhibition of ¹⁴C-IAA transport in petiole segments isolated 1 or 2 days after treatment. Using 100 micromolar TDZ, the inhibition was detectable 19 hours after treatment and was complete by 27 hours. Both leaves and petiole segments exhibited a marked increase in ethylene production following treatment with TDZ at concentrations of 0.1 micromolar or greater. The involvement of ethylene in this TDZ response was evaluated by examining the effects of two inhibitors of ethylene action: silver thiosulfate, 2,5-norbornadiene. One day after treatment, both inhibitors effectively antagonized the TDZ-induced inhibition of auxin transport. Two days after TDZ treatment both inhibitors were ineffective. The decrease in IAA transport in TDZ treated tissues was associated with increased metabolism of IAA. The transport of ¹⁴C-2,4-dichlorophenoxyacetic acid was also inhibited by TDZ treatment. This inhibition was not accompanied by increased metabolism. Incorporation of TDZ into the receiver blocks had no effect on auxin transport. The ability of the phytotropin N-1-naphthylphthalamic acid to stimulate IAA uptake from a bathing medium was reduced in TDZ-treated tissues. This reduction is thought to reflect a decline in the auxin efflux system following TDZ treatment.     

Effect of thidiazuron, a cytokinin-active urea derivative, in cytokinin-dependent ethylene production systems

Cytokinins are known to stimulate ethylene production in mungbean hypocotyls synergistically with indoleacetic acid (IAA), in mungbean hypocotyls synergistically with Ca²⁺, and in wilted wheat leaves. Thidiazuron, a substituted urea compound, mimicked the effect of benzyladenine (BA) in all three systems. In the Ca²⁺ + cytokinin system and the IAA + cytokinin systems of mungbean hypocotyls, thiadiazuron was slightly more active than BA at equimolar concentration. In mungbean hypocotyls exogenously applied IAA was rapidly conjugated into IAA asparate, and this conjugation process was effectively inhibited by thidiazuron, as by cytokinins. In the wilted wheat leaves system, 10 micromolar thidiazuron exerted stress ethylene production equal to that exerted by 1 millimolar BA, indicating that thidiazuron is more active than BA by two orders. The structure-activity relationship of thidiazuron and its thiadiazolylurea analogs in stimulating Ca²⁺-dependent ethylene production in mungbean hypocotyls was found to agree well with the structure-activity relationship of these derivatives in promoting the growth of callus tissues. These results indicate that thidiazuron and its derivatives are highly active to mimic the adenine-type cytokinin responses in promoting ethylene production and that the structure-activity relationship in promoting the growth of callus and in promoting ethylene production is similar.     

Ethylene involvement in vegetative bud formation in tobacco thin layers

Summary The role of ethylene in vegetative bud regeneration was studied in cultured tobacco (Nicotiana tabacum L. cvSamsun) thinlayer expiants. The experimental approach consisted in supplementing the bud-inducing medium with an inhibitor of ethylene biosynthesis, aminoethoxyvinylglycine (AVG), an ethylene antagonist, silver thiosulphate (STS), or an ethylene-releasing compound, 2-chloroethylphosphonic acid (CEPA), at various concentrations. The organogenic response was assessed both macroscopically (percentage of bud-forming expiants, final number of buds per expiant) and cytohistologically (number, characteristics, and localisation of meristemoids and bud primordia). The time course of ethylene production during culture was also evaluated. At the end of culture (day 27) all the expiants treated with these compounds had a lower number of buds compared to controls. STS was detrimental to meristemoid initiation at all the concentrations tested. In contrast, 0.5 M AVG, which strongly inhibited ethylene production, provoked a large increase in the formation of meristemoids early in culture and the appearance of anomalous (twin) buds. CEPA reduced meristemoid formation but, at the lower concentrations (1 and 10 M) speeded up bud emergence. On the whole it mainly favoured disorganised growth and xylogenesis. The results of this work highlight the contrasting effects of ethylene in relation to the two critical stages of the organogenic process, i.e., meristemoid formation and bud primordium development.Abbreviations ACC 1-aminocyclopropane-1-carboxylic acid - AVG aminoethoxyvinylglycine - STS silver thiosulphate - CEPA 2-chloroethylphosphonic acid - IAA indole-3-acetic acid - BA benzyladenine - HF hormone-free     

Ethylene biosynthesis in Regnellidium diphyllum and Marsilea quadrifolia

The pathway of ethylene biosynthesis was examined in two lower plants, the semi-aquatic ferns Regnellidium diphyllum Lindm. and Marsilea quadrifolia L. As a positive control for the ethylene-biosynthetic pathway of higher plants, leaves of Arabidopsis thaliana (L.) Heynh. were included in each experiment. Ethylene production by Regnellidium and Marsilea was not increased by treatment of leaflets with 1-aminocyclopropane-1-carboxylic acid (ACC), the precursor of ethylene in higher plants. Similarly, ethylene production was not inhibited by application of aminoethoxyvinylglycine and -aminoisobutyric acid, inhibitors of the ethylene biosynthetic enzymes ACC synthase and ACC oxidase, respectively. However, ACC was present in both ferns, as was ACC synthase. Compared to leaves of Arabidopsis, leaflets of Regnellidium and Marsilea incorporated little [¹⁴C]ACC and [¹⁴C]methionine into [¹⁴C]ethylene. From these data, it appears that the formation of ethylene in both ferns occurs mainly, if not only, via an ACC-independent route, even though the capacity to synthesize ACC is present in these lower plants.Abbreviations ACC 1-aminocyclopropane-1-carboxylic acid - AdoMet S-adenosyl-l-methionine - AIB -aminoisobutyric acid - AVG aminoethoxyvinylglycine This research was supported by the U.S. Department of Energy through grant No. DE-FG02-91ER20021 and, in part, by a fellowship of the National Engineering and Research Council of Canada to Jacqueline Chernys.