Ethylene Production, 1-Aminocyclopropane-l-Carboxylic Acid Content and Its Conversion to Ethylene in Axial Segments of Dormant and Nondormant Cocklebur Seeds

During the imbibition of water, the change in the ethylene productionof axial segments of nondormant (ND) cocklebur (Xanthium pennsylvanicumWallr.) seeds paralleled the change in the content of free 1-aminocyclopropane-l-carboxylicacid (ACC), but not the change in conjugated hydrolysable ACCin the axes. Aminoethoxyvinylglycine, anoxia and a-aminoisobutyricacid inhibited ethylene production, the lattertwo compoundscausing the accumulation of free ACC. Administration of ACCgreatly enhanced ethylene production in the axes. Thus, freeACC seems to be a direct precursor of ethylene production inthe axial tissue of cocklebur seeds. Imbibed dormant (D) axes characterized by inferior ethyleneproduction had less ability to convert exogenously applied ACCto ethylene as compared to ND axes. But, there was little differencebetween the D and ND axes in the endogenous contents of freeand conjugated ACC. This suggests that the inferior ethyleneproduction found in detached D axes is associated with the lowactivity of an ACC-ethylene converting system. (Received December 17, 1982; Accepted April 30, 1983)     

Release of heat pretreatment-induced dormancy in lettuce seeds by ethylene or cytokinin in relation to the production of ethylene and the synthesis of 1-aminocyclopropane-1-carboxylic acid during germination

The germination of lettuce (Lactuca sativa L.) seeds was greatly reduced when the seeds were heated at 97°C for 30 h prior to imbibition. This dormancy was effectively released when ethylene (1–100 ppm) or benzyladenine (BA) (0.005–0.05 mM) was applied during the imbibition period. Ethylene was not required during the early part of imbibition, but was essential during the period immediately prior to radicle protrusion. Treatment with 1-aminocyclopropane-1-carboxylic acid (ACC) (0.1–10 mM) stimulated germination, but was not as effective as ethylene or cytokinin treatment. During the germination of nondormant lettuce seeds, ethylene production increased rapidly and reached a peak at 24 h, which coincided with the emergence of the radicle, and then declined; the level of ACC increased as ethylene production rate increased, but remained at a high level after radicle protrusion. In heat-pretreated dormant lettuce seeds, the increases in percent germination, ethylene production, and ACC levels were all delayed and lower than those of nondormant seeds, and these increases were accelerated by treatment with ethylene or cytokinin.     

Changes in Ethylene Production and in 1-Aminocyclopropane-1-carboxylic Acid (ACC) and Malonyl-ACC Contents of Cocklebur Seeds during Their Development

Ethylene production in developing cocklebur (Xanthium pennsyluanicumWallr.) seeds peaked when the dry weight of the seeds beganto increase in the early period of development. The productionthen began to decrease and stopped when the dry weight increasewas completed. The upsurge of ethylene production in the earlydevelopmental period paralleled increases in ACC synthase activityand the 1-aminocyclopropane-1-carboxylic acid (ACC) contentof the seeds, both of which rapidly decreased later. Malonyl-ACC (MACC) accumulated in developing cocklebur seedsduring the early period of development, before the ACC contentand ethylene production increased. Although the ACC synthaseactivity, ACC content and ethylene production showed markeddecreases, the MACC content remained almost unchanged duringthe middle period of seed development, with a pronounced decreaseoccurring in the late period. Exogenous application of MACCdid not promote ethylene production of seeds collected at thelate developmental stage. Aminoethoxyvinylglycine, an inhibitorof ACC synthase, strongly inhibited the ethylene productionof the same lot of seeds. Therefore, the decrease in the MACCcontent in developing cocklebur seeds was not due to reuse ofMACC for ethylene production. (Received May 24, 1984; Accepted August 15, 1984)     

Phytochrome Control of Cellulase Activity in Datura ferox L. Seeds and its Relationship with Germination

The influence of phytochrome on endosperm softening and cellulaseactivity was studied on light-stimulated Datura ferox seeds.Endosperm softening preceded the earliest signs of radicle protrusion,and there was good correlation between the % of seeds with softendosperm at 48 h after R and germination at 96 h after R. Cellulaseactivity was stimulated by R and the increase in activity preceded,by more than 24h, radicle protrusion and endosperm softening.The effect of R was reversed by FR, but, by delaying the irradiationwith FR until cellulase activity had increased significantly,it was observed that removing Pfr did more than just stop anyfurther increase, the level of cellulase activity decreasedin about 24 h close to the dark controls. Cellulase activitywas decreased by a FR irradiation even when more than 60% germinationhad escaped from reversion. These results indicate that phytochromeinfluence on cellulase is not an indirect consequence of thestimulus of germination and that the continuous presence ofPfr is required for the cellulase activity to remain high. Thepossibility that cellulase and other degrading enzymes may bepart of the mechanism of light-induced germination is discussed. Key words: Phytochrome, germination, cellulase     

ACC conversion to ethylene by sunflower seeds in relation to maturation,germination and thermodormancy

Conversion of exogenous 1-aminocyclopropane-1-carboxylic acid (ACC) to ethylene was studied in sunflower (Helianthus annuus L., cv. Mirasol) seeds in relation to germinability. Ethylene production from ACC decreased during seed maturation, and non-dormant mature seeds were practically unable to synthesize ethylene until germination and growth occurred, indicating that ethylene forming enzyme (EFE) activity developed during tissue imbibition and growth. ACC conversion to ethylene was reduced by the presence of pericarp, and in young seedlings it was less in cotyledons than in growing axes.ACC conversion to ethylene by cotyledons from young seedlings was optimal at c. 30°C, and was strongly inhibited at 45°C. Pretreatment of imbibed seeds at high temperature (45°C) induced a thermodormancy and a progressive decrease in EFE activity.Abscisic acid and methyl-jasmonate, two growth regulators which inhibit seed germination and seedling growth, and cycloheximide were also shown to inhibit ACC conversion to ethylene by cotyledons of 3-day-old seedlings and by inbibed seeds.Abbreviations ABA abscisic acid - ACC 1-aminocyclopropane-1-carboxylic acid - CH cycloheximide - EFE ethylene forming enzyme - IAA indole-3-acetic acid - Me-Ja methyl-jasmonate     

Ethylene Production in Pea and Cocklebur Seeds of Differing Vigour

Relationships between seed vigour and ethylene (C₂H₄) productionwere studied using C₂H₄-responsive fatty cocklebur seeds (Xanthiumpennsyhanicum Wallr.) and C₂H₄-insensitive starchy pea seeds(Pisum sativum L. cv. Alaska), which had been harvested in differentyears and subjected to different storage conditions. In bothspecies, the seeds with the highest vigour evolved the largestamounts of C₂H₄ during a period of water imbibition. The reductionof C₂H₄ production in cocklebur seeds occurred concomitantlywith the reduction in the growth potentials of both axial andcotyledonary tissues. Similarly, the activity of ACC-C₂H₄ conversionincreased with soaking, and was greater in seeds of high vigourcompared with those of low vigour. However, the change in ACCcontent in pea seeds differed from that in cocklebur seeds.That is, pea seeds with high vigour accumulated less ACC duringan imbibition period than those with low vigour. From theseresults it was suggested that the inferior C₂H₄ production bylow vigour pea seeds is mainly attributable to low ACC-C₂H₄conversion, whereas that by low vigour cocklebur seeds is dueto the shortage of ACC supply in addition to the reduced ACC-C₂H₄conversion. However, germination of deteriorated cocklebur seedswas not restored by exposure to ACC or C₂H₄, suggesting thatthe loss of seed vigour reduces the responsiveness of seedsto C₂H₄ as well as C₂H₄ production. Key words: Pea, cocklebur, seed vigour, ethylene production, 1-aminocyclopropane-1-carboxylic acid     

Characterization of ethylene/gibberellic acid control of germination in Lactuca sativa L.

Ethylene production during germination of lettuce seeds (Lactucasativa L., cv. Premier Great Lakes) occurred at two distinctlydifferent rates. A very low rate of ethylene release was observedprior to the 12th hour of incubation at 22?C. The rate of ethyleneproduction, however, increased 100 fold between the 12th and16th hour of incubation. This high rate of ethylene productiononly occurred in the presence of seeds which exhibited a visibleprotrusion of the radicle. The duration of exposure to a supraoptimaltemperature (32?C) was inversely proportional to the percentgermination at 32?C. Ethylene production and growth were notblocked by incubating visibly germinated seeds at 32?C. Exogenous ethylene partially restored germination at 32?C, butonly in the light. Gibberellic acid partially substituted forthe induced light requirement but not for ethylene. It was concludedthat the supraoptimal temperature raised the threshold concentrationof ethylene required for germination. This threshold requirementwas satisfied in the presence of exogenous ethylene. Germinationat 32?C was abo dependent upon the presence of GA. With exogenousethylene present, the GA-mediated system was presumably reinstatedor bypassed by exposing the seeds to either light or GA. Theinitial low rate of ethylene production apparently regulatessubsequent germination but only when present at a minimum thresholdconcentration. Those events initiating germination have obviouslyoccurred prior to the time of radicle emergence. Post-germinationethylene production, therefore, did not break thermodormancy,but occurred simultaneously with radicle emergence. (Received November 29, 1976; )     

Effect of short-chain fatty acids on the ethylene pathway in embryonic axes of Cicer arietinum during germination

The effect of short-chain saturated fatty acids (C₅–C₁₀) on the biosynthesis of ethylene in embryonic axes of chick-pea ( Cicer arietinum L.) seeds was investigated. The emergence of radicle and fresh weight of embryonic axes diminished with increasing number of carbons. The inhibition of germination caused by lower concentrations (1 m M ) of fatty acids (C₅–C₁₀) was partially reversed by exogenous 1-aminocyclopropane-1-carboxylic acid (ACC), whereas exogenous ethylene was able to overcome the inhibitory effect provoked by all concentrations (1–5 m M ) of applied fatty acids (C₅–C₁₀). Ethylene production rates, and enzyme activities of ACC synthase and ACC oxidase decreased concomitantly with the molecular mass and increasing concentration of fatty acids. The inhibitory effect of these acids on ethylene production seems to result not only from a decreased ACC synthesis, but also from an enhancement of 1-malonylamino)cyclopropane-1-carboxylic acid (MACC) synthesis.     

Ethylene biosynthesis and strigol-induced germination of Striga asiatica

Germination of witchweed [ Striga asiatica (L.) Kuntze], an important parasite on cereal crops, is stimulated by several natural and synthetic compounds. In the present study the role of ethylene in germination of Striga asiatica in response to strigol was examined. Unconditioned seeds and those conditioned for 3 days produced negligible amounts of ethylene in response to strigol. However, extending the conditioning period to 5 and 8 days increased ethylene evolution by more than 10-fold. Ethylene production preceded radicle protrusion and was detectable within 3h after treatment. No germination was observed in the first 6 h of exposure to strigol. Germination and ethylene production increased with strigol concentration. Strigol-induced germination was considerably reduced by the ethylene action inhibitors. 2. 5-norbornadiene, silver thiosulphate and CO₂. The ethylene precursor 1-aminocyclopropane-1-carboxyac acid (ACC) at 5 to 200 μ M elicited neither germination nor ethylene production. However, a combination of strigol and ACC resulted in a high germination rate and copious ethylene production. Both germination and ethylene production were reduced by CoCl₂ and cyclobeximide, inhibitors of the ethylene-forming enzyme and of protein synthesis, respectively. The results are consistent with a model in which conditioning and strigol are required to remove a restriction on the ethylene biosynthetic pathway and in which the ethylene-forming enzyme is rate limiting.     

Ethylene promotes ethylene biosynthesis during pea seed germination by positive feedback regulation of 1-aminocyclo-propane-1-carboxylic acid oxidase

 Increased ethylene evolution accompanies seed germination of many species including Pisum sativum L., but only a little is known about the regulation of the ethylene biosynthetic pathway in different seed tissues. Biosynthesis of the direct ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC), the expression of ACC oxidase (ACO), and ethylene production were investigated in the cotyledons and embryonic axis of germinating pea seeds. An early onset and sequential induction of ACC biosynthesis, accumulation of Ps-ACO1 mRNA and of ACO activity, and ethylene production were localized almost exclusively in the embryonic axis. Maximal levels of ACC, Ps-ACO1 mRNA, ACO enzyme activity and ethylene evolution were found when radicle emergence was just complete. Treatment of germinating seeds with ethylene alone or in combination with the inhibitor of ethylene action 2,5-norbornadiene showed that endogenous ethylene regulates its own biosynthesis through a positive feedback loop that enhances ACO expression. Accumulation of Ps-ACO1 mRNA and of ACO enzyme activity in the embryonic axis during the late phase of germination required ethylene, whereas Ps-ACS1 mRNA levels and overall ACC contents were not induced by ethylene treatment. Ethylene did not induce ACO in the embryonic axis during the early phase of germination. Ethylene-independent signalling pathways regulate the spatial and temporal pattern of ethylene biosynthesis, whereas the ethylene signalling pathway regulates high-level ACO expression in the embryonic axis, and thereby enhances ethylene evolution during seed germination. Received: 28 September 1999 / Accepted: 27 December 1999     

Chilling-Induced Ethylene Production in Relation to Chill-Sensitivity in Phaseolus spp.

Guye, M. G, Vigh, L. and Wilson, J. M. 1987. Chilling-inducedethylene production in relation to chill-sensitivity in Phaseolusspp.—J. exp. Bot. 38: 680–690. Ethylene production from the primary leaves of six bean (Phaseolusspp.) cultivars known to differ in chill-sensitivity, was monitoredat 23 ?C following chilling of whole plants at 5 ?C for 24 h.The more chill-tolerant cultivars produced greater amounts ofchilling-induced ethylene than the chill-sensitive cultivars.The onset of maximum ethylene production rates and the followingdecline in rates was more rapid in chill-tolerant cultivars.This pattern of ethylene production was also similar when chill-tolerancewas chemically enhanced by choline treatment. The low levelsof ethylene production in chill-sensitive genotypes was alsoreflected by their poor ability to convert the exogenously appliedethylene precursor, 1-aminocyclopropane-l-carboxylic acid (ACC),to ethylene. Moderate levels of leaf water deficit induced by chilling chill-tolerantcultivars and choline treated plants appeared to stimulate chilling-inducedethylene production. High levels of leaf wilt, shown by morechill-sensitive cultivars, reduced this stimulatory effect.Ethylene production was slightly greater when warming was carriedout in the light rather than in the dark. Key words: Ethylene, ACC, choline, chill-sensitivity, Phaseolus     

Induction of 1-Aminocyclopropane-1-carboxylic Acid Synthase in Wounded Mesocarp Tissue of Winter Squash Fruit and the Effects of Ethylene

1-Aminocyclopropane-1-carboxylic acid (ACC) synthase activityincreased rapidly after wounding of mesocarp tissue of wintersquash fruit (Cucurbita maxima Duch.) and reached a peak at16 h after excision and then declined sharply. The rise in ACCsynthase activity was followed by increases in the endogenousACC content and the rate of ethylene production. The activityof ethylene forming enzyme (EFE) also increased rapidly in theexcised discs of mesocarp of winter squash fruit. ACC synthase activity was strongly inhibited by aminoethoxyvinylglycinewith a Ki value of 2.1 µM. Michaelis-Menten constant ofACC synthase for S-adenosylmethionine was 13.3 µM. Ethylene suppressed the induction of ACC synthase in the woundedmesocarp tissue. The suppression by ethylene increased withthe increasing concentrations of applied ethylene and the maximumeffect was obtained at about 100 µl 1^–1 ethylene,at which point the induction was suppressed by 54%. Ethylenedid not inhibit ACC synthase activity, nor did it suppress theinduction of EFE, but rather it slightly enhanced the latter. (Received August 24, 1984; Accepted October 29, 1984)     

Polyamine contents, ethylene synthesis, and BrACO2 expression during turnip germination

Contents of total free [PA(S)] and conjugated polyamines [PA(SH), PA(PH)] were higher in turnip (Brassica rapa L. cv. Rapa) seeds during imbibition (0–36 h) and radicle protrusion (36–48 h) than during the further growth (10 d). Ethylene production was activated with the protrusion, reaching a maximum at the second day of germination and dropping afterwards. The application of ethrel accelerated radicle emergence but the direct intervention of ethylene in the breaking of the seed coat was not clear from the use of ethylene-biosynthesis inhibitors (CoCl₂ and AVG). Finally, in this work the gene BrACO2 was characterized. Although its expression was not detected in seeds through zygotic embryogenesis, it increased concomitantly with the germination process.     

Ethylene, Nitrate and Nitrite Interactions in the Promotion of Dark Germination of Common Purslane Seeds

Ethylene (10 µ1^–1) caused about one-third of highlydark-dormant seeds of common purslane (Portulaca oleracea L.)to germinate in the dark. Attempts were made to increase germinationin the dark with nitrate and ethylene combinations. When applieddirectly to the seeds, KNO₃ did not stimulate germination andKNO₃ plus ethylene did not increase germination above that ofethylene alone. Pre-incubation of seeds in KNO₃ for 4 to 7 dbefore the ethylene applications significantly increased germination.The effects of the KNO₃ pre-incubation were additive at eachof four ethylene concentrations (0.1–100 µ11^–1).Potassium nitrate was effective only when ethylene followedthe KNO₃ pre-incubation period. Potassium nitrite stimulatedabout 25 per cent of the seeds to germinate without a pre-incubationperiod and without ethylene. Also, ethylene plus KNO₂ enhancedgermination above that achieved by either stimulus alone. Silvernitrate did not block the ethylene promotion of germination,but reversed the typical ethylene inhibition of seedling growthfollowing germination. The results support the views that nitrateexerted its effect via conversion to nitrite within the seedand that the rate of nitrate conversion may be a limiting factorin the dark germination of common purslane seeds. Ethylene mayfacilitate nitrite activity by increasing seed sensitivity tothe stimulus. Common purslane, Portulaca oleracea L., ethylene, nitrate, nitrite, germination, dormancy     

Induction by Electric Currents of Ethylene Biosynthesis in Cucumber (Cucumis sativus L.) Fruit

The effects of an electric current on ethylene biosynthesis were investigated in cucumber (Cucumis sativus L.) fruit that were producing almost no ethylene. Direct currents at 0.5 to 3.0 milliamperes induced much ethylene synthesis, with a rapid continuous increase in the rate, which reached a peak within 5 to 6 hours and then decreased. The rate of production was greater with a stronger current. Ethylene production was not observed after the use of a sine-wave alternating current (60 hertz) at 3 milliamperes, the magnitude at which a direct current had the greatest effect. The activity of 1-aminocyclopropane-1-carboxylic acid (ACC) synthase and ethylene forming enzyme (EFE) increased before the rise in ethylene production. ACC synthase and EFE were activated sixfold and fourfold, respectively, by 2 hours. The concentration of ACC increased linearly up to 6 hours and then decreased. Ethylene induction by an electric current was suppressed almost completely by the infiltration of the cucumbers with 5 millimolar aminooxyacetic acid, an inhibitor of ACC synthase, and was also suppressed 70% by 5 millimolar salicylic acid, an inhibitor of EFE. The results indicate that the ethylene induced by the direct current was synthesized via the ACC-ethylene pathway as a result of electrical stress, a new kind of stress to be identified.     

Effect of Lithium on Thigmomorphogenesis in Bryonia dioica Ethylene Production and Sensitivity

Rubbing internodes of Bryonia dioica plants reduced their ethylene production but increased their capacity to convert 1-aminocyclopropane-1-carboxylic acid (ACC) to ethylene. These results were explained by the previously shown rubbing-induced decrease of indoleacetic acid, which controls the level of ACC synthase, and by the increase of membrane-associated peroxidases which would participate in the conversion of ACC-ethylene. Pretreatment of the plants with Li had no significant effect on control plants but counteracted the rubbing-induced decrease of ethylene production and diminished the capacity of the internodes to convert ACC to ethylene. Exogenously applied ethylene induced an increase of peroxidase activity similar to that caused by rubbing. Inasmuch as both effects were reduced by Li, it was concluded that Li inhibition of thigmomorphogenetic processes was essentially due to a Li inhibition of the effect of ethylene formed in response to mechanical stimuli. The decreased ethylene production and ACC conversion capacity in the presence of Li were explained by a cellular redistribution of peroxidases.     

The Respiratory Metabolism of Strelitzia juncea Ait. Seeds: The Effect of Dormancy Release through Oxygen Incubation of the Seeds on the Activity of Glucose-6-Phosphate Dehydrogenase and 6-Phosphogluconate Dehydrogenase

Incubation of Strelitzia juncea seeds in an oxygen atmosphereresulted in an increase in the G6PDH activity of crude embryoextracts on day one, while radicle protrusion started on dayfive. Similarly, 6PGDH activity increased over the first 4 dof incubation in oxygen. The ratio of 6PGDH/G6PDH was 3.0<x < 3.7 regardless of treatment or incubation period. Supplying oxygen per se to dormant seed and studying its effecton the activity of the two key pentose phosphate (PP) pathwayenzymes, appear to support Roberts' hypothesis that oxygen shortagerestricts PP pathway activity in dormant seeds. Key words: Dormancy, pentose phosphate pathway, Strelitzia juncea     

Effects of Exogenous 1,3-Diaminopropane and Spermidine on Senescence of Oat Leaves : II. Inhibition of Ethylene Biosynthesis and Possible Mode of Action

The effects of the polyamines spermidine and 1,3-diaminopropane on ethylene biosynthesis and chlorophyll (Chl) loss were studied in peeled leaves of oat (Avena sativa L., var. Victory) incubated in the dark. Peeling off the epidermal cells induces an increase in 1-aminocyclopropane-1-carboxylate (ACC) synthase activity, resulting in an enhanced ACC and ethylene formation. Both polyamines inhibit ethylene biosynthesis from methionine by inhibiting ACC synthase activity and, more effectively, the conversion of ACC to ethylene. They also inhibit Chl loss occurring between 24 and 48 h of dark incubation; but, as shown by inhibitor experiments, inhibition of Chl loss does not result from inhibition of ethylene formation. Ethylene production and Chl loss, both associated with senescence, require membrane integrity; thus, treatments which promote deterioration of membranes inhibit both processes. Ca²⁺ in the incubation medium competitively reduces the polyamine-mediated inhibition of ACC conversion and Chl loss. The data suggest that polyamines initially attach to membranes, thereby inducing changes which, in turn, lead to inhibition of ethylene biosynthesis and retardation of senescence.     

Effects of exogenous ethylene on ethylene production in citrus leaf tissue

Exogenous ethylene stimulated ethylene production in intact citrus (Citrus sinensis L. Osbeck cv. “Washington Navel”) leaves and leaf discs following a 24-hour exposure. Studies with leaf discs showed that ethylene production decreased when ethylene was removed by aeration. The extent of stimulation was dependent upon the concentration of exogenous ethylene (1-10 microliters per liter). Silver ion blocked the autocatalytic effect of ethylene at concentrations of 0.5 millimolar and lower, but increased ethylene production at higher concentrations. The stimulating effect of ethylene resulted from the enhancement of both 1-aminocyclopropane-1-carboxylic acid (ACC) formation and the conversion of ACC to ethylene. Whereas autocatalysis was evident following 24 hours incubation, autoinhibition of wound- and mannitol-induced ethylene production was observed during the first 24-hour incubation. Ethylene treatment during this period resulted in a marked decrease in ACC levels and ethylene production rates. Furthermore, in leaf discs treated for 24 hours with ethylene, ethylene production rates increased greatly during the first 2 hours after removal of exogenous ethylene by aeration. This increase was eliminated if the discs were transferred to propylene instead of air, indicating that the autocatalytic effect of ethylene is counteracted by its autoinhibitory effect. It is suggested that autocatalysis involves increased synthesis of ACC synthase and the enzyme responsible for the conversion of ACC to ethylene, whereas autoinhibition involves suppression of the activity of these two enzymes.