Effect of l-aminocyclopropane-l-carboxylic acid,silver nitrate,and norbornadiene on plant regeneration from maize callus cultures

The effect of the ethylene antagonists norbornadiene and silver nitrate and the ethylene precursor l-aminocyclopropane-l-carboxylic acid (ACC) on Zea mays plant regeneration was studied. A 12-fold increase in plant regeneration, as measured by number of plants obtained per gram fresh weight from callus cultures of maize inbreds Pa91 and H99, was obtained by 250 M norbornadiene and 100 M silver nitrate treatments. An increase in amout of nonregenerable tissue and a 68% decrease in plant regeneration were associated with callus treated with 1 mM ACC. Ethylene emanation from 1 mM ACC treated callus reached a maximum of 170 nl g^–1 h^–1 after 3 days compared to 7 nl g^–1 h^–1 for the control. The free proline content was up to 80% lower in 1 mM ACC treated callus grown for 30 days on medium with or without 12 mM proline, respectively, as compared to each control. These studies indicate that ethylene action inhibitors such as norbornadiene and silver nitrate can be used to increase plant regeneration efficiency from maize callus cultures.Abbreviations ACC l-aminocyclopropane-l-carboxylic acid - gfw gram fresh weight     

Role of ethylene in chlorophyll degradation in radish cotyledons

The effect of age of radish seedlings on changes in chlorophyll concentration caused by ethylene was examined. Ethylene was produced at 2–4 nl g^–1 h^–1 following excision of cotyledons from 5-to 20-day-old seedlings. The youngest cotyledons maintained this rate, whereas ethylene synthesis declined by as much as 80% during a 24-h period in older cotyledons. The youngest cotyledons continued to accumulate chlorophyll in the dark, but after 7 days cotyledons lost chlorophyll and the proportion of chlorophyll lost increased with age. Ethylene promoted, and norbornadiene inhibited, this loss of chlorophyll; in combined treatments the effects of ethylene and norbornadiene were competitive. The maximal rate of chlorophyll loss occurred in 1l L^–1 ethylene; extrapolation of the response to concentration indicated that half-maximum loss would occur at 0.005–0.01 l L^–1 ethylene. In cotyledons from 20-day-old seedlings, chlorophyll degradation occurred mainly after 24 h from excision and transfer to the dark. Chlorophyll degradation during 48 h in the dark was affected by norbornadiene or ethylene applied from 0–24 h or from 24–48 h.     

The role of ethylene in the senescence of carnation flowers, a review

The senescence of flower petals is a highly regulated developmental process which requires active gene expression and protein synthesis. The biochemical changes associated with petal senescence in carnation flowers include an increase in hydrolytic enzymes, degradation of macro-molecules, increased respiratory activity and a climacteric-like increase in ethylene production. It is clear that the gaseous phytohormone ethylene plays a critical role in the regulation and coordination of senescence processes. Many reviews on physiology and mode of action of ethylene are available. Molecular cloning led to the isolation of genes involved in ethylene biosynthesis and action. This review describes the current status of the studies on regulation of ethylene biosynthesis and ethylene response in carnation flowers. An overview is given of studies on senescence-related gene expression and possibilities to improve postharvest longevity by genetic engineering.Abbreviations ACC 1-aminocyclopropane-1-carboxylic acid - AIB -amino-isobutyric acid - AOA amino oxyacetic acid - AVG aminoathoxyvinyl glycine - DACP diazocyclopentadiene - EFE ethylene forming enzyme - MACC malonyl 1-aminocyclopropane-1-carboxylic acid - MTA 5-methylthio-adenosine - NBD 2,5 norbornadiene - ppb parts per billion - SAM S-adenosyl-methionine - STS silver thiosulphate     

Altered sensitivity to ethylene in ‘Tardivo’, a late‐ripening mutant of Clementine mandarin

‘Tardivo’ mandarin is a mutant of ‘Comune’ Clementine with a delay in peel degreening and coloration, allowing late harvesting. In this work, we have explored if the late‐harvesting phenotype of ‘Tardivo’ mandarin is related to altered perception and sensitivity to ethylene. The peel degreening rate was examined after a single ethephon treatment or during a continuous ethylene application in fruits at two maturation stages. In general, ethylene‐induced peel degreening was considerably delayed and reduced in fruits of ‘Tardivo’, as well as the concomitant reduction of chlorophyll (Chl) and chloroplastic carotenoids, and the accumulation of chromoplastic carotenoids. Analysis of the expression of genes involved in Chl degradation, carotenoids, ABA, phenylpropanoids and ethylene biosynthesis revealed an impairment in the stimulation of most genes by ethylene in the peel of ‘Tardivo’ fruits with respect to ‘Comune’, especially after 5 days of ethylene application. Moreover, ethylene‐induced expression of two ethylene receptor genes, ETR1 and ETR2, was also reduced in mutant fruits. Expression levels of two ethylene‐responsive factors, ERF1 and ERF2, which were repressed by ethylene, were also impaired to a different extent, in fruits of both genotypes. Collectively, results suggested an altered sensitivity of the peel of ‘Tardivo’ to ethylene‐induced physiological and molecular responses, including fruit degreening and coloration processes, which may be time‐dependent since an early moderated reduction in the responses was followed by the latter inability to sustain ethylene action. These results support the involvement of ethylene in the regulation of at least some aspects of peel maturation in the non‐climacteric citrus fruit.     

Mechanism of naphthaleneacetic Acid conjugation: no effect of ethylene

Formation of naphthaleneacetic acid-glucose (NAGLu) in detached leaves, floating on α-naphthaleneacetic acid-1-¹⁴C (NAA, 0.05 microcurie per milliliter, 3.1 μm)-buffer solution (phosphate-citrate, pH 4.2) began immediately while there was a 2- to 4-hour lag before NAA-asparatate (NAAsp) could be detected. Subsequent increase in the NAAsp conjugate reflected a decrease in free NAA to 1 to 2% of the total radioactivity taken up. Pretreatment with 31 μm¹²C-NAA for 18 hours doubled NAAsp formation after transfer for 4 hours to ¹⁴C-NAA. Pretreatment with ethylene, as ethephon (up to 400 milligrams per liter) or ethylene gas (10 microliters per liter), did not induce NAAsp formation. In the presence of NAA, ethylene had no effect on NAA conjugation. Similarly, CO₂ (5%) did not modify the formation of the conjugates. Rhizobitoxine (1.87 μm) inhibited NAA-induced ethylene production but did not prevent NAA-induced formation of NAAsp. We concluded that the conjugation of NAA with aspartic acid is not mediated by ethylene.     

Indole-3-acetic acid concentration and ethylene evolution during early fruit development in peach

Ethylene evolution was measured from greenhouse-grown Jerseyglo peach fruits beginning 29 days after anthesis. Indole-3-acetic acid (IAA) levels were measured in the pericarp and seed tissues of individual fruits on a single shoot when variable ethylene evolution was noted. Despite hand-pollinating all flowers on the same day, variability within the shoot existed in fruit fresh weight, IAA levels, and ethylene evolution. Seed IAA concentration increased as fruit and seed fresh weight increased and ranged from 106 to 1572 ng. g^–1. As pericarp fresh weight increased, IAA levels in this tissue decreased. Ethylene evolution rates ranged from 0.21 to 1.07 nl. g.^–1 h^–1 and were not correlated with IAA concentration in seed, pericarp, or the whole fruit. High rates of ethylene evolution from the whole fruit occurred prior to increased IAA concentration in the seed.Fruits were excised from field-grown Redskin peach trees beginning 40 days after full bloom. Fruits from field sampled shoots appeared to be more physiologically advanced than the greenhouse-grown Jerseyglo fruits. Pericarp IAA concentration was low, ranging from 2.8 to 6.5 ng. g^–1. Seed concentrations accounted for 75% of the IAA found in the fruit and ranged from 239 to 1042 ng. g^–1. As with greenhouse-grown samples, whole fruit IAA concentration tended to decrease as fruits increased in fresh weight.     

Ethylene as promoter of wheat grain maturation and ear senescence

This work was aimed at testing the involvement of ethylene in the maturation of grain and senescence of the foliar structures of the wheat inflorescence. Whole wheat ears emitted ethylene to the atmosphere. From pre-anthesis, ethylene emission progressively increased from 0.76 nl g^–1FW h^–1 to a peak 1.53 nl g^–1FW h^–1 at the hard dough stage of the grains, to fall to a minimum of 0.10 nl g^–1FW h^–1 at the dormant seed stage. Ethephon increased the ethylene release, hastened the process of grain maturation and senescence of the ears. Aminoethoxyvinylglycine and silver thiosulfate produced the opposite effects. It is concluded that ethylene plays a role in grain maturation and in the senescence of the green bracts of the inflorescence.Abbreviations Ag⁺ = silver ion - AVG = aminoethoxyvinylglycine - SAM = S-adenosylmethionine - STS = silver thiosulfate - TGW = thousand grain weight Consejo Nacional de Investigaciones Científicas y Tecnológicas (CONICET)     

Role of ethylene in chlorophyll degradation in radish cotyledons

The effect of age of radish seedlings on changes in chlorophyll concentration caused by ethylene was examined. Ethylene was produced at 2–4 nl g^?1 h^?1 following excision of cotyledons from 5-to 20-day-old seedlings. The youngest cotyledons maintained this rate, whereas ethylene synthesis declined by as much as 80% during a 24-h period in older cotyledons. The youngest cotyledons continued to accumulate chlorophyll in the dark, but after 7 days cotyledons lost chlorophyll and the proportion of chlorophyll lost increased with age. Ethylene promoted, and norbornadiene inhibited, this loss of chlorophyll; in combined treatments the effects of ethylene and norbornadiene were competitive. The maximal rate of chlorophyll loss occurred in 1μl L^?1 ethylene; extrapolation of the response to concentration indicated that half-maximum loss would occur at 0.005–0.01 μl L^?1 ethylene. In cotyledons from 20-day-old seedlings, chlorophyll degradation occurred mainly after 24 h from excision and transfer to the dark. Chlorophyll degradation during 48 h in the dark was affected by norbornadiene or ethylene applied from 0–24 h or from 24–48 h.     

The stimulation of cell extension by ethylene and auxin in aquatic plants

Elongation of the shoots of three aquatic plants (Hydrocharis morsus-ranae, Regnellidium diphyllum and Ranunculus sceleratus) is stimulated by treatment with ethylene or IAA. The effects of the two hormones are additive, and experiments with an ethylene biosynthesis inhibitor and silver ions indicate that the mechanisms by which ethylene and IAA stimulate growth may be different. Hydrocharis and Ranunculus leaf discs synthesize [¹⁴C]ethylene from [¹⁴C]methionine, but no [¹⁴C]ethylene is formed by Regnellidium, suggesting the existence of an alternative pathway of ethylene biosynthesis in the fern.Abbreviations IAA Indole-3-acetic acid - RBA 1,2-amino-4-(2-aminoethoxy)-trans-3-butenoic acid     

The effects of 2,2′-bipyridine and other metal chelators on the conversion of 1-aminocyclopropane-1-carboxylic acid to ethylene in rice

Effects of metal chelators, 2,2-bipyridine, 8-hydroxyquinoline and 1,10-phenenthroline, on the conversion of 1-aminocyclopropane-1-carboxylic acid (ACC) to ethylene in detached leaves of light-grown rice (Oryza sativa) seedlings and detached shoots of etiolated rice seedlings were investigated. Metal chelators strongly inhibited the in vivo ACC oxidase activity in detached leaves and detached etiolated shoots. This inhibition could be partially recovered by Fe²⁺. Our results support the notion that Fe²⁺ is an essential cofactor for the conversion of ACC to ethylene in vivo.Abbreviations ACC 1-aminocyclopropane-1-carboxylic acid - BP 2,2-bypyridine - HQ 8-hydroxylquinoline - MJ methyl jasmonate - PA 1,10-phenanthroline - Put putrescine     

Characterization of ethylene production in developing strawberry fruit

Ethylene production, ACC content, and ACC oxidase activity were determined in strawberry fruit harvested at different stages of development and in fruit harvested green and developed in vitro in solutions containing sucrose. In fruit harvested at progressive stages of development from green through full ripe, ethylene production and ACC oxidase activity decreased whereas ACC content increased between the white and pink stages. Fruit detached at the green stage and developed to full ripe by immersion of the cut pedicel in sucrose solutions exhibited an increase in ACC content, decreased ethylene production, and no change in ACC oxidase activity. Detached green fruit provided with sucrose containing 0.5 mM silver (STS) had elevated ethylene production and more ACC oxidase activity than did fruit incubated without the silver salt. Green fruit provided with sucrose containing 1 mM ACC showed markedly increased ACC content, ACC oxidase activity, and ethylene production. These increases were noted following 4 days incubation in ACC, and were more pronounced after 11 days, at which time fruit of all treatments had attained a full-ripe stage of development. Calyx tissue exhibited more ACC oxidase activity, less ACC content, and similar ethylene production compared with receptacle tissue. ACC synthase could not be detected in fruit harvested at different developmental stages or in fruit detached and developed in vitro.abbreviations ACC 1-aminocyclopropane-1-carboxylic acid - HQS 8-hydroxyquinoline hemisulfate - SAM S-adenosyl methionine - STS silver thiosulfate     

Effect of 1-methylcyclopropene and methylenecyclopropane on ethylene binding and ethylene action on cut carnations

1-Methylcyclopropene (1-MCP), formerly designated as Sis-X, has been shown to be an effective inhibitor of ethylene responses in carnation flowers in either the light or the dark. The binding appears to be to the receptor and to be permanent. A 6 h treatment at 2.5 nl l^–1 is sufficient to protect against ethylene, and 0.5 nl l^–1 is sufficient if exposure is for 24 h. As carnation flowers age, a little higher concentration appears to be needed. Most of the natural increase in ethylene production during senescence is prevented by treatment with 1-MCP. A closely related compound, methylenecyclopropane shows ethylene activity. A tritium labelled 1-MCP (60 mCi mmol^–1) has been prepared. A higher specific activity is needed for more critical studies.     

Abscisic acid- and ethylene-induced defoliation of Radermachera sinica L.

Radermachera sinica L. is an ornamental plant with demonstrated sensitivity to ethylene-induced leaf abscission. In this study, we examine the relationship between abscisic acid (ABA) and ethylene in initiating the abscission response. Treatment with 1 l L^\s-1 of ethylene, 1 mM 1-aminocyclopropane-1-carboxylic acid (ACC) or 1 mM ABA resulted in complete defoliation of leaf explants. Application of 0.125 mM silver thiosulfate (STS) inhibited ethylene- and ACC-induced abscission but had no effect on explants treated with ABA. The ABA-induced abscission was unaffected by treatment with aminoethoxyvinylglycine (AVG) or aminooxyacetic acid (AOA). Treatment of explants with 1 mM cobalt chloride (CoCl₂) or 2000 l L^\s-1 of norbornadiene (NBD) completely inhibited abscission in explants treated with 1 l L^\s-1 ethylene or 1 mM ACC but they were only marginally effective in blocking ABA-induced abscission despite the lower level of endogenous ethylene. ABA appeared to increase the sensitivity of explants to ethylene. However, the evidence suggests that ABA may also function independent of ethylene to induce leaf abscission in R. sinica.Abbreviations ABA abscisic acid - ACC 1-aminocyclopropane-1-carboxylic acid - AOA aminooxyacetic acid - AVG aminoethoxyvinylglycine - CoCl₂ cobalt chloride - NBD norbornadiene - STS silver thiosulfate     

Involvement of ethylene in chlorophyll degradation in peel of citrus fruits

The effect of ethylene on chlorophyll degradation in the peel of Robinson tangerine (X Citrus reticulata Blanco) and calamondin (X Citrofortunellamitis [Blanco] Ingram and Moore) fruits was studied. The chlorophyll degrading system in the peel of these two citrus species was not self-sustaining but required ethylene to function. Chlorophyll degradation ceased immediately when fruit were removed from ethylene and held in ethylene-free air at 0.2 atmospheric pressure. However, at atmospheric pressure, chlorophyll degradation continued for 24 hours in the absence of exogenous ethylene. Although chlorophyllase levels were negatively correlated with chlorophyll content in the peel (r = −0.981; P < 0.01), the level of chlorophyllase activity did not change when fruit were removed from ethylene, even though chlorophyll degradation had stopped. From these observations, it was concluded that ethylene is necessary for chlorophyll degradation in the two species of citrus studied, but its primary role is not solely for the induction of chlorophyllase activity.     

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     

Enhanced plant regeneration in pearl millet (Pennisetum americanum) by ethylene inhibitors and cefotaxime

Cefotaxime, a cephalosporin antibiotic, and different ethylene inhibitors, such as silver nitrate, cobalt chloride, nickel chloride and O-acetyl salicylic acid, significantly delayed the loss of regeneration potential in embryogenic cultures of Pennisetum americanum. In the presence of these chemicals, ethylene content in the atmosphere of the culture vessel was less than that of the control. Cefotaxime, silver nitrate and O-acetyl salicyclic acid did not have any effect on callus growth based on fresh weight, while growth based on dry weight was enhanced by O-acetyl salicyclic acid.Abbreviations ASA O-acetyl salicylic acid - BA benzyladenine - CW coconut water - 2,4-D 2,4-dichlorophenoxyacetic acid - IAA indole-3-acetic acid - NAA -naphthaleneacetic acid - MS Murashige and Skoog     

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     

Effects of 1-MCP on the vase life and ethylene response of cut flowers

Pretreatment for 6 h with low concentrations of 1-MCP (1-Methylcyclopropene, formerly designated as SIS-X), a cyclic ethylene analog, inhibits the normal wilting response of cut carnations exposed continuously to 0.4 l·l^–1 ethylene. The response to 1-MCP was a function of treatment concentration and time. Treatment with 1-MCP was as effective in inhibiting ethylene effects as treatment with the anionic silver thiosulfate complex (STS), the standard commercial treatment. Other ethylene-sensitive cut flowers responded similarly to carnations. In the presence of 1 l·l^–1 ethylene, the vase life of 1-MCP-treated flowers was up to 4 times that of the controls.Abbreviations 1-MCP 1-Methylcyclopropene - STS silver thiosulfate     

Role of ethylene in the senescence of detached rice leaves

The role of ethylene in the senescence of detached rice leaves in relation to their changes in 1-aminocyclopropane-1-carboxylic acid (ACC) content and ethylene production was studied. In freshly excised rice leaf segments, ACC level and ethylene production rates were very low. Following incubation, the rates of ethylene production increased and reached a maximum in 12 h, and subsequently declined. The rise of ethylene production was associated with a 20- to 30-fold increase in ACC level.

Ethylene seems to be involved in the regulation of the senescence of detached rice leaves. This conclusion was based on the observations that (a) maximum ethylene production preceded chlorophyll degradation, (b) ACC application promoted chlorophyll degradation, (c) inhibitors of ethylene production and ethylene action retarded chlorophyll degradation, and (d) various treatments such as light, cycloheximide, α,α-dipyridyl, Ni²⁺, and cold temperature, which retarded chlorophyll degradation, also inhibited ethylene production.

Abscisic acid promoted senescence but significantly decreased ethylene production, whereas benzyladenine retarded senescence but promoted ethylene production. This is interpreted to indicate that abscisic acid treatment increased the tissue sensitivity to ethylene, whereas benzyladenine treatment decreased it.