Ethylene production and metabolism of 1-aminocyclopropane-1-carboxylic acid inChenopodium rubrum L. as influenced by photoperiodic flower induction

Chenopodium rubrum plants, induced to flower by three cycles of 12 h darkness and 12 h light, produced 42% less ethylene than vegetative plants kept under continuous light. Plants that had each dark cycle broken by 2 h light in the middle did not flower and produced almost as much ethylene as the vegetative plants. Shoots and roots of plants of all three experimental treatments had a similar content of 1-aminocyclopropane-1-carboxylic acid (ACC), the mean amounting to about 2 nmol · g^–1 dry weight. Also the content of N-malonyl-ACC (MACC) was similar in shoots of all three treatments. MACC content in roots was shown to be much higher, especially in the treatments with three dark periods (about 85 nmol · g^–1 dry weight). When labeled [2,3-¹⁴C] ACC was administered, the relative contents of ACC and MACC were very similar among all three treatments. The only process influenced by flower induction was ACC conversion to ethylene. Induced plants converted 36% less ACC than the vegetative ones. Plants subjected to night-break converted almost as much ACC to ethylene as vegetative plants. It is concluded that flower induction in the short-day plantChenopodium rubrum decreases ethylene production by decreasing their capability of converting ACC to ethylene.     

Transport and Metabolism of 1-Aminocyclopropane-1-carboxylic Acid in Sunflower (Helianthus annuus L.) Seedlings

Transport and metabolism of [2,3-¹⁴C] 1-aminocyclopropane-1-carboxylic acid (ACC) from roots to shoots in 4-day-old sunflower (Helianthus annuus L.) seedlings were studied. [¹⁴C]ACC was detected in, and ¹⁴C₂H₄ was evolved from, shoots 0.5 hours after [¹⁴C]ACC was supplied to roots. Ethylene emanation from the shoots returned to normal levels after 6 hours. The roots showed a similar pattern, although at 24 hours ethylene emanation was still slightly higher than in those plants that did not receive ACC. [¹⁴C]N-malonyl-ACC (MACC) was detected in both tissues at all times sampled. [¹⁴C]MACC levels surpassed [¹⁴C]ACC levels in the shoot at 2 hours, whereas [¹⁴C]MACC levels in the root remained below [¹⁴C]ACC levels until 6 hours, after which they were higher. Thin-layer chromatography analysis identified [¹⁴C] ACC in 1-hour shoot extracts, and [¹⁴C]MACC was identified in root tissues at 1 and 12 hours after treatment. [¹⁴C]ACC and [¹⁴C] MACC in the xylem sap of treated seedlings were identified by thin-layer chromatography. Xylem transport of [¹⁴C]ACC in treated seedlings, and transport of ACC in untreated seedlings, was confirmed by gas chromatography-mass spectrometry. Some evidence for the presence of [¹⁴C]MACC in xylem sap in [¹⁴C]ACC-treated seedlings is presented. A substantial amount of radioactivity in both ACC and MACC fractions was detected leaking from the roots over 24 hours. A second radiolabeled volatile compound was trapped in a CO₂-trapping solution but not in mercuric perchlorate. Levels of this compound were highest after the peak of ACC levels and before peak MACC levels in both tissues, suggesting that an alternate pathway of ACC metabolism was operating in this system.     

Changes m Ethylene Production in Relation to l-Aminocyclopropane-l-Carboxylic Acid and Its Malonyl Conjugate in Waterlogged Wheat Plants

When wheat seedlings were subjected to waterlogging, 1-aminocyelopropane-l-carboxylic acid (ACC), an ethylene precursor, accumulated in large quantity in roots. In shoots, ACC and ethylene production also increased, but declined with the prolonged periods of waterlogging. However, ACC content in roots maintained in high level during the whole period of waterlogging. Drainage caused a drastic drop in both ACC content and ethylene production in waterlogged plants to control level. 1-(malonylamino) cyclopropane-l-carboxylic acid (MACC) level in roots subjected to waterlogging showed little changes. However, MACC content in shoots kept increasing during the 9-days period of waterlogging. At later period of waterlogging (longer than 5 days) when ACC and ethylene production bad dropped, the. level of MACC continued to increase. Draining stopped this increasing, but did not reduced its level. When exogenous ACC was introduced into the leaves via transpiration stream, the ability of leaves of waterlogged plant to convert ACC to MACC was much higher than control. The data presented showed that at the later stage of waterlogging, the conversien of a great quantity of ACC to MACC in waterlogged wheat plants is the cause of the reduction of ethylene production and ACC content. It was suggested that the formation of MACC is another way of regulation in ethylene biosynthesis. Among leaves of different ages, the enhancement of ethylene, ACC and MACC content was more pronounced in older leaves than in younger laves during the waterlogging period. The physiological significance of adaptation to waterlogging stress was discussed.     

Diurnal Fluctuations in Ethylene Formation in Chenopodium rubrum

Ethylene formation was studied in 5- to 6-d-old Chenopodium rubrum seedlings under the following light regimes: continuous light (CL), continuous darkness (CD), and alternating light/darkness (12 h of each). No significant regular oscillations in ethylene formation were found in either the CL or CD groups. In the light/dark regime, pronounced diurnal fluctuations in ethylene formation were observed. Activity of 1-aminocyclopropane-1-carboxylic acid (ACC) oxidase was transiently increased on transfer from light to dark and vice versa. In CL, ACC oxidase activity did not change significantly, whereas in CD, it decreased continuously after the initial increase. The in vivo levels of ACC and N-malonyl-ACC (MACC) were constant for the first few hours of darkness, then decreased dramatically, but increased again in the light. In constant darkness, the level of ACC displayed endogenous rhythm, with minimum values at h 12 and 44, and a maximum value at h 32 to 36. The level of MACC in both shoots and roots decreased in the CD group until h 12, and then remained constant until h 30 before decreasing continuously. We conclude that the photoperiodic regime affects both ACC and MACC levels, as well as the conversion of ACC to ethylene. Correlation of the described changes in ethylene formation to photoperiodic flower induction is discussed.     

Activity of Ageing Carnation Flower Parts and the Effects of 1-(Malonylamino)cyclopropane-1-Carboxylic Acid-Induced Ethylene

Peak levels of 1-aminocyclopropane-l-carboxylic acid (ACC) in flower parts of ageing carnations (Dianthus caryophyllus L. cv Scanea 3C) were detected 6 to 9 days after flower opening. The ethylene climacteric and the first visible sign of wilting was observed 7 days after opening. The concentration of conjugated ACC in these same tissues peaked at day three with reduction of 70% by day 4. From day 5 to day 9 all parts followed a diurnal pattern of increasing in conjugate levels 1 day and decreasing the next. Concentrations of conjugated ACC were significantly higher than those of ACC in all ageing parts. Preclimacteric petals treated with ACC or 1-(malonylamino)-cycloprane-1-carboxylic acid (MACC), started to senesce 30 to 36 hours after treatment. When petals were treated with MACC plus by 0.1 millimolar aminoethyoxyvinylglycine, premature senescence was induced, while ethylene production was suppressed relative to MACC-treated petals. Petals treated with MACC and silver complex produced ethylene, but did not senesce. The MACC-induced ethylene was inhibited by the addition of 1.0 millimolar CoC1₂. These results demonstrate MACC-induced senescence in preclimacteric petals. The patterns of ACC and MACC detected in the flower parts support the view that an individual part probably does not export an ethylene precursor to the remainder of the flower inducing senescence.     

Identification and Metabolism of 1-(Malonylamino)cyclopropane-1-carboxylic Acid in Germinating Peanut Seeds

Peanut seeds (Arachis hypogea L. Yue-you 551) contain 50 to 100 nanomoles per gram conjugated 1-aminocyclopropanecarboxylic acid (ACC). Based on paper chromatography, paper electrophoresis, and gas chromatography-mass spectrometry, it was verified that the major ACC conjugate was N-malonyl-ACC (MACC). Germinating peanut seeds converted [2-¹⁴C]ACC to ethylene 70 times more efficiently than N-malonyl-[2-¹⁴C]ACC; when ACC was administered, most of it was metabolized to MACC. Germinating peanut seeds produced ethylene and converted l-[3,4-¹⁴C]methionine to ethylene; this ethylene biosynthesis was inhibited by aminoethoxyvinylglycine. These data indicate that MACC occurs in peanut seeds but does not serve as the source of ethylene during germination; ethylene is, however, synthesized from methionine via ACC.     

Effect of Darkness on Growth and Flowering of Chenopodium rubrum and C. murale Plants in vitro

Chenopodium rubrum, a short-day plant, and C. murale, a long-day plant, were grown in vitro in continuous darkness. Control C. rubrum plants exposed to continuous darkness for 15 d at cotyledonary phase, did not flower, while 80 % of plants flowered on the medium with 5 % glucose and 10 mg dm⁻³ GA₃. Control C. murale plants exposed to continuous darkness for 10 d at the age of 4^th pair of leaves, did not flower, while GA₃ (1 – 5 mg dm⁻³) stimulated flowering up to 65 %. This revised version was published online in July 2006 with corrections to the Cover Date.     

The Conversion of 1-(Malonylamino)cyclopropane-1-Carboxylic Acid to 1-Aminocyclopropane-1-Carboxylic Acid in Plant Tissues

Since 1-(malonylamino)cyclopropane-1-carboxylic acid (MACC), the major conjugate of 1-aminocyclopropane-1-carboxylic acid (ACC) in plant tissues, is a poor ethylene producer, it is generally thought that MACC is a biologically inactive end product of ACC. In the present study we have shown that the capability of watercress (Nasturtium officinale R. Br) stem sections and tobacco (Nicotiana tabacum L.) leaf discs to convert exogenously applied MACC to ACC increased with increasing MACC concentrations (0.2-5 millimolar) and duration (4-48 hours) of the treatment. The MACC-induced ethylene production was inhibited by CoCl₂ but not by aminoethoxyvinylglycin, suggesting that the ACC formed is derived from the MACC applied, and not from the methionine pathway. This was further confirmed by the observation that radioactive MACC released radioactive ACC and ethylene. A cell-free extract, which catalyzes the conversion of MACC to ACC, was prepared from watercress stems which were preincubated with 1 millimolar MACC for 24 hours. Neither fresh tissues nor aged tissues incubated without external MACC exhibited enzymic activity, confirming the view that the enzyme is induced by MACC. The enzyme had a K_m of 0.45 millimolar for MACC and showed maximal activity at pH 8.0 in the presence of 1 millimolar MnSO₄. The present study indicates that high MACC levels in the plant tissue can induce to some extent the capability to convert MACC to ACC.     

The quantification of 1-(malonylamino)cyclopropane-1-carboxylic acid in plant tissues by quantitative mass spectrometry

A method for the quantitation of 1-(malonylamino)cyclopropane-1-carboxylic acid (MACC), a conjugated form of 1-aminocyclopropane-1-carboxylic acid (ACC), in plants is described. [2,2,3,3-²H₄]MACC has been used as an internal standard for selected ion monitoring/isotope dilution quantitation of MACC in wheat seedlings and in tomato leaves. This method is compared with a widely-used two step indirect assay for MACC, which is based upon hydrolysis of MACC to ACC and conversion of ACC by hypochlorite reagent to ethylene which is subsequently quantified by gas chromatography.     

Effect of plant growth regulators on ethylene production, 1-aminocyclopropane-1-carboxylic acid oxidase activity, and initiation of inflorescence development of pineapple

With the development of pineapple [Ananas comosus (L.) Merr.] as a fresh fruit crop, it became common to force inflorescence development with ethephon [(2-chloroethyl)phosphonic acid] or ethylene throughout the year. Environmental induction (EI) of inflorescence development disrupts scheduling of fruit harvest and may cause significant losses if small plants are induced, resulting in fruits that are too small to be marketable. Our objective was to identify plant growth regulators (PGRs) that could inhibit EI. Because circumstantial evidence indicates that EI occurs in response to naturally produced ethylene or changes in plant sensitivity to it, most work was done with PGRs that inhibit ethylene biosynthesis or block ethylene action. The synthetic auxin 2-(3-chlorophenoxy)propionic acid (CPA) was included because in one study it reduced the percentage of EI. GA₃, aminooxyacetic acid (AOA), aminoethoxyvinylglycine (AVG), daminozide [butanedioic acid mono-(2,2-dimethylhydrazide)], and silver thiosulfate (STS) had no effect on EL CPA, paclobutrazol [(2RS,3RS)-1-(4-chlorophenyl)methyl-4,4-dimethyl-2(1h-1,2,4-triazol-1-yl)penten-3-ol], and uniconazole [(E)-(p-chlorophenyl)-4,4-dimethyl-2-(1,2,4-triazol-1-yl)-1-penten-3-ol] delayed or inhibited EI of pot-grown pineapple plants. Uniconazole and paclobutrazol inhibited growth and ethylene production by leaf basal-white tissue, and either or both effects could account for the inhibition of EI. Production of 1-aminocyclopropane-1-carboxylic acid (ACC) was unaffected by these compounds, but the activity of ACC oxidase, which converts ACC to ethylene, was inhibited and probably accounts for the reduced ethylene production by leaf basal-white tissue. CPA stimulated ethylene production by stem apical tissue approximately fourfold relative to the control. ACC oxidase activity and the malonyl-ACC (MACC) content in stem apical tissue were also greater than in the control, indicating that CPA greatly stimulated the production of ACC and its sequestration into MACC. The mechanism by which CPA delayed or inhibited EI is not known. CPA, paclobutrazol, and uniconazole appear to have some potential for inhibiting EI of pineapple. Their effect on yield needs to be determined.Abbreviations ACC oxidase 1-aminocyclopropane-1-carboxylic acid oxidase - CPA 2-(3-chlorophenoxy)propionic acid - AOA aminooxyacetic acid - AVG aminoethoxyvinylglycine - daminozide butanedioic acid mono-(2,2-dimethylhydrazide) - DM dry mass - ethephon [(2-chloroethyl)phosphonic acid] - FM fresh mass - GA gibberellin - EI environmental induction of inflorescence development - IA inflorescence appearance - LSD Fisher's protected least significant difference - MACC malonyl-ACC - NAA naphthaleneacetic acid - PGR plant growth regulator - paclobutrazol (2RS,3RS)-1-(4-chlorophenyl)methyl-4,4-dimethyl-2-(1h-1,2,4-triazol-1-yl)penten-3-ol] - uniconazole (E)-(p-chlorophenyl)-4,4-dimethyl-2-(1,2,4-triazol-1-yl)-1-penten-3-ol - STS silver thiosulfate - M-leaf fourth leaf - Ml-L first leaf younger than M-leaf     

Accumulation of 1-(malonylamino)cyclopropane-1-carboxylic acid in ethylene-synthesizing tobacco leaves

During the hypersensitive reaction of Samsun NN tobacco to tobacco mosaic virus (TMV) the inoculated leaves synthesize large quantities of ethylene. At the same time, 1-(malonylamino)cyclopropane-1-carboxylic acid (MACC), a conjugate of the ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC) accumulates. Smaller amounts of MACC are formed concomitant with ethylene synthesis during the normal development of tobacco leaves. The conjugate appears neither to be hydrolysed to liberate ACC, nor to be transported to other plant parts. Its accumulation thus reflects the history of the operation of the pathway of ethylene synthesis in the leaf. In floating leaf discs exogenously applied ACC was converted only slowly to both ethylene and MACC. More ethylene and less MACC were produced in darkness than in light, suggesting that environmental conditions may influence the ratio at which ACC in converted to either ethylene or MACC.     

Effects of pH and heavy metal concentrations in solution culture on the proton release,growth and elemental composition ofAlyssum murale andRaphanus sativus L.

The proton release by a species that can hyperaccumulate nickel (Alyssum murale) and by a non-accumulator (Raphanus sativus L.) was studied at different pH and heavy metal concentrations in solution culture. Both factors influenced the growth and composition of the plants.A. murale was more sensitive than radish to a decrease of pH from 7.0 to 6.0 in the growth medium; plant yield and proton production diminished with decreasing pH. However, yields and proton production of radish only decreased at pH 5.5. The differences in the amounts of protons produced between the hyperaccumulator species and radish were not large enough to conclude that decreasing pH in the rhizosphere ofA. murale is a mechanism for heavy metal solubilization.Nickel concentrations inA. murale followed the typical pattern of an accumulator plant — more Ni was accumulated in the shoots than in the roots. Lower concentrations of Zn and Cd occurred in the shoots than in roots ofA. murale, and also of Ni in radish. The concentrations of Co inA. murale shoots were increased when Zn, Ni and Cd were absent from the nutrient solution. However, Co concentrations in radish shoots were independent of the concentrations of other heavy metals in the growth medium.     

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.     

Promotion ofin vitro shoot formation from excised roots of silktree (Albizzia julibrissin) by an oxime ether derivative and other ethylene inhibitors

Summary This report describes the regeneration response of excised seedling roots of silktree (Albizzia julibrissin) to added ethylene precursors/generators (1-amino-cyclopropane-1-carboxylic acid [ACC], 2-chloroethylphosphonic acid [CEPA]), biosynthesis inhibitors (aminoethoxyvinylglycine [AVG], an oxime ether derivative [OED={[(ispropylidene)-amino]oxy}-acetic acid-2-(methoxy)-2-oxoethyl ester], CoCl² [Co⁺⁺]), and an ethylene action inhibitor (AgNO₃ [Ag⁺]). When placed on B5 medium, about 50% of the control explants formed shoot buds within 15 days. Addition of ACC or CEPA (1–10 µM) to the culture medium decreased both the percentage of cultures forming shoots and the number of shoots formed per culture. In contrast, AVG and OED (1–10 µM) increased shoot formation to almost 100% and increased the number of shoots formed per culture. Likewise, both Co⁺⁺ and Ag⁺ (1–10 µM) increased shoot regeneration, but the number of shoots produced after 30 days was less than with AVG or OED. The inhibitors of ethylene biosynthesis were partially effective in counteracting the inhibitory effect of ACC on shoot formation. These results suggest that modulation of ethylene biosynthesis and/or action can strongly influence the formation of adventitious shoots from excised roots of silktree.Abbreviations ACC 1-aminocyclopropane-1-carboxylic acid - AVG aminoethoxyvinylglycine - CEPA 2-chloroethylphosphonic acid - OED oxime ether derivative     

Effects of arbuscular mycorrhizal fungi on tree growth, leaf water potential, and levels of 1-aminocyclopropane-1-carboxylic acid and ethylene in the roots of papaya under water-stress conditions

 Seedlings of papaya (Carica papaya L. var. Solo) were transplanted to pots with or without an arbuscular mycorrhizal (AM) fungus (Gigaspora margarita Becker and Hall). After 3 months, half the plants were subjected to water stress by withdrawing irrigation. The leaf water potential (LWP) was measured during 20 days of water-stress treatment and then the plants were harvested. Root ethylene and 1-aminocyclopropane-1-carboxylic acid (ACC) concentrations were measured and plant fresh weight determined. The LWP decreased during the water-stress treatment and this decrease was more severe in the non-AM plants. Plant fresh weight was higher for AM than non-AM plants under both conditions. Under well-irrigated conditions, the ethylene concentration in the roots was increased by the presence of AM, although there was no significant difference between AM and non-AM roots in ACC levels. ACC increased in both AM and non-AM roots under water-stress conditions. The water-stress treatment resulted in a marked increase in ethylene concentration in non-AM roots but the concentration in AM roots was slightly lower than under normal conditions. Accepted: 7 July 2000     

Ethylene and auxin-ethylene interaction in adventitious root formation in mung bean (Vigna radiata) cuttings

The role of ethylene in adventitious root formation and its involvement in auxin-induced rooting were investigated in cuttings ofVigna radiata (L.). Treatment with 30 M indole-3-acetic acid (IAA) for 24 h slightly inhibited rooting, whereas the same concentration of indole-3-butyric acid (IBA) significantly stimulated it. Ethylene derived from 1-aminocyclopropane-1-carboxylic acid (ACC) increased the number of adventitious roots but inhibited their emergence and elongation. Endogenous levels of ethylene, ACC, and malonyl-ACC (MACC) were initially higher in cuttings treated with IAA. This trend was quickly reversed, and cuttings, particularly hypocotyls, treated with IBA produced higher levels of ethylene and had more ACC and MACC during most of the rooting process. Aminoethoxyvinylglycine significantly inhibited rooting, but its inhibitory effect could not be reversed by ACC. The data suggest that the stimulating effect of IBA on rooting is closely associated with its induction of ACC and ethylene biosynthesis.     

Influence of the triazole growth retardant BAS 111..W on phytohormone levels in senescing intact pods of oilseed rape

Foliar treatment of oilseed rape plants (Brassica napus L.ssp. napus cv. Linetta) with the growth retardant BAS 111..W at the 5th leaf stage delayed pod senescence during early maturation. Changes of immunoreactive cytokinin- and abscisic acid (ABA)- like substances and of the ethylene precursor 1-aminocyclo-propane-1-carboxylic acid (ACC) and its malonyl-conjugate (MACC) were determined in intact whole pods. When compared with control plants, higher levels of total chlorophyll correlated with four-fold and three-fold increases of trans-zeatin riboside- and dihydrozeatin riboside-type cytokinins, respectively, in the pods of plants treated with 0.25 mg BAS 111..W per plant. Isopentenyladenosine-type cytokinins and ACC and MACC contents remained virtually unchanged, whereas ABA levels dropped considerably below those of controls (60% reduction). However, when analysed at late pod maturity, BAS 111..W treatment no longer affected the total chlorophyll content, or the levels of cytokinins, ABA, ACC and MACC. We hypothesize that the retardant-induced changes in the hormonal status of the pods, favouring the senescence-delaying cytokinins as opposed to abscisic acid, could contribute to the developmental delay.     

Circadian Ethylene Synthesis in <Emphasis Type="Italic">Sorghum bicolor</Emphasis>: Expression and Control of the System at the Whole Plant Level

Ethylene production by sorghum is rhythmic and the amplitude of the rhythm is increased both by dim, far-red enriched light and in mutant plants deficient in phytochrome B. The mechanisms involved in controlling ethylene production were examined in detail by measuring the rate of ethylene production among organs and tissues, examining the organ-specific levels of ACC (1-aminocyclopropane-1-carboxylic acid, the ethylene precursor) and investigating the contribution of the roots to shoot ethylene production. The results demonstrate that the expanding leaves were the major source of ethylene under dim, far-red enriched light and in the phytochrome B mutant. Enhanced ethylene production by the expanding leaf appeared to be the result of targeted delivery of ACC to this tissue. Root ACC levels were much higher than those in the shoot but roots converted much less of this endogenous ACC to ethylene. Applying ACC to the roots had only a marginal effect on their ethylene production, but greatly increased that of the shoots. Decapitated shoots continued to produce ethylene in a rhythmic pattern but the amplitude decreased with time compared to intact plants. The results collectively suggest that some, but not all, of the shoot ethylene rhythm depends on the transport of ACC from the roots to the shoots.