Role of aminocyclopropane-l-carboxylic acid in ethylene release by distal tissues following localized application of ethephon in Cucurbita pepo

The application of ethephon to a single leaf of Cucurbita pepo L. cv. Trailing Marrow plants caused a huge increase in ethylene production from the treated organ and an increased rate of ethylene production from other parts of the plant. These increases were particularly marked in the shoot apex and expanding leaf. Prior treatment with aminoethoxyvinylglycine (AVG), an ethylene biosynthesis inhibitor, blocked the increased production of ethylene at sites distant from the point of ethephon application. This strongly suggests that the increased ethylene production at these distant sites is due to ethylene biosynthesis and not a result of the translocation of ethylene released by the breakdown of ethephon at the site of application. Assays of 1-aminocyclopropane-l-carboxylic acid (ACC), an ethylene precursor, showed that it increased substantially after ethephon application but was at undetectable levels in the presence of AVG. It is proposed that the application of ethephon stimulates ethylene biosynthesis, but that transport through the plants is effected by ACC which is then converted to ethylene at the shoot apex and leaves.     

Role of 1-aminocyclopropane-1-carboxylic acid in the control of female flowering in Cucurbita pepo

Cucurbita pepo L. cv. Trailing Marrow is monoecious, bearing separate male and female flowers and the first functional flowers are usually male. Treatment with 300 ppm ethephon delayed and greatly reduced male flower production and also increased female flower numbers. When plants were sprayed with aminoethoxyvinylglycine (AVG) no female flowers were produced but male flower production was unaffected. Even when ethephon was applied to AVG-treated plants there was still complete inhibition of female flower production. Similarly, AVG-treated plants subsequently exposed to 4000 ppm ethylene for two days never produced female flowers. AVG inhibits the penultimate stage in ethylene biosynthesis i.e. immediately before 1-aminocyclopropane-1-carboxylic acid (ACC). Although spraying AVG-treated plants with ACC did not reverse the inhibition, application of ACC via a cut petiole for a 72 h period following AVG application did cause female flowers to form. The evidence indicates that ACC and not ethylene is the factor controlling female flower production in C. pepo .     

Internode length in Pisum. Two further mutants, lh and ls, with reduced gibberellin synthesis, and a gibberellin insensitive mutant, lk

Three further internode length mutants in peas ( Pisum sativum L.), lh , ls and lk , were examined to determine if they influenced gibberellin synthesis or sensitivity. Two mutants, lh and ls , showed pronounced elongation in response to applied GA₁ and extracts from their shoots contained little gibberellin-like activity when assayed on the rice seedling (cv. Tan ginbozu) bioassay compared with similar extracts from essentially isogenic Lh and Ls plants. The third mutant, lk , was almost insensitive to applied GA₁ and at no dose rate did it become a phenocopy of normal Lk plants. Extracts from the shoots of lk and Lk segregants contained similar levels of gibberellinlike substances. All three mutants influenced growth in both the light and the dark, although only the effect of genes Lh and Ls were graft transmissible. These results suggest that lh and ls are mutants with reduced gibberellin synthesis, while lk is the first gibberellin-insensitive dwarfing gene identified in peas.     

Influence of growth retardants on the internode and ethylene production of sunflower plants

The growth-retarding activity of the norbornenodiazetine tetcyclacis and the di-oxanylalkenyl triazole LAB 150 978 as well as the ethylene-forming compounds 2-chloroethyl-phosphonic acid (ethephon) and 1-amino-cyclopropane-l-carboxylic acid (ACC) on stem histogenesis and ethylene production of sunflower plants ( He-lianthus annuus L. cv.Spanners Allzweck) has been studied. The shoot growth of plants hydroponically grown and treated was reduced by the compounds. The shortening in the length of the 1st internode caused by tetcyclacis and LAB 150 978 was mainly induced by inhibition of cell division (the internode possessed fewer cortical cells per cell file). In contrast, ethephon and ACC decreased internode elongation mainly by reducing the rate of cell enlargement.
The ethylene production of sunflower seedlings cultivated on agar nutrient medium rose with increasing concentrations of ethephon and ACC, the shoot length of the plants being progressively reduced.
Tetcyclacis and LAB 150 978 inhibited both the formation of ethylene and shoot growth. It is suggested that in contrast to ethephon and ACC, tetcyclacis and LAB 150 978 do not achieve their growth-retarding effect by influencing the production of ethylene.     

Gravitropism in Higher Plant Shoots : IV. Further Studies on Participation of Ethylene

Ethylene at 1.0 and 10.0 cubic centimeters per cubic meter decreased the rate of gravitropic bending in stems of cocklebur (Xanthium strumarium L.) and tomato (Lycopersicon esculentum Mill), but 0.1 cubic centimeter per cubic meter ethylene had little effect. Treating cocklebur plants with 1.0 millimolar aminoethoxyvinylglycine (AVG) (ethylene synthesis inhibitor) delayed stem bending compared with controls, but adding 0.1 cubic centimeter per cubic meter ethylene in the surrounding atmosphere (or applying 0.1% ethephon solution) partially restored the rate of bending of AVG-treated plants. Ethylene increases in bending stems, and AVG inhibits this. Virtually all newly synthesized ethylene appeared in bottom halves of horizontal stems, where ethylene concentrations were as much as 100 times those in upright stems or in top halves of horizontal stems. This was especially true when horizontal stems were physically restrained from bending. Ethylene might promote cell elongation in bottom tissues of a horizontal stem or indicate other factors there (e.g. a large amount of `functioning' auxin). Or top and bottom tissues may become differentially sensitive to ethylene. Auxin applied to one side of a vertical stem caused extreme bending away from that side; gibberellic acid, kinetin, and abscisic acid were without effect. Acidic ethephon solutions applied to one side of young seedlings of cocklebur, tomato, sunflower (Helianthus annuus L.), and soybean (Glycine max [L.] Merr.) caused bending away from that side, but neutral ethephon solutions did not cause bending. Buffered or unbuffered acid (HCl) caused similar bending. Neutral ethephon solutions produced typical ethylene symptoms (i.e. epinasty, inhibition of stem elongation). HCl or acidic ethephon applied to the top of horizontal stems caused downward bending, but these substances applied to the bottom of such stems inhibited growth and upward bending—an unexpected result.     

Requirement for the action of endogenous ethylene during germination of non-dormant seeds of Amaranthus caudatus

The role of endogenous ethylene during germination of non-dormant seeds of Amaranthus caudatus L. was investigated. The seeds readily germinated in water and darkness at 24°C. Application of ethylene or of its precursor I-aminocyclopropane-I-carboxylic acid (ACC) slightly increased the rate of germination. Both compounds effectively antagonized osmotic inhibition by polyethyleneglycol. Application of aminoethoxyvinylglycine (AVG) reduced ethylene production by 90% but did not inhibit germination. However, germination was inhibited by 2,5-norbornadiene, a competitive inhibitor of ethylene action. This inhibition was counteracted by ethylene, ethephon or ACC and enforced by AVG. It is concluded that the action of endogenous ethylene is an indispensable factor during germination of non-dormant seeds of A. caudatus. Ethylene action is required from the start of imbibition on. In water, low levels of endogenous ethylene are sufficient for this action. PEG increased the ethylene requirement considerably.     

Internode length in Pisum. Two further gibberellin-insensitivity genes, lka and lkb

Two further internode length genes are identified in Pisum sativum L. and named lka (identified from line NGB5865) and lkb (from NGB5862). These genes result in a similar phenotype, which includes reduced stem elongation, peduncle length and basal branching, and 'banding' of the stem. These effects are similar to, but less severe than, those of gene lk . Genes lka and lkb influence gibberellin (GA) sensitivity, since mutants NGB5865 and NGB5862 possess similar levels of endogenous GA-like substances to the wild-type parental cultivar Torsdag and respond less to applied GA₁ than do wild-type plants or GA-synthesis mutants of a similar stature. The action of genes lka and lkb is localised in the young apical tissue but is not thought to involve GA-perception, since plants possessing genes lka and lkb are not true phenocopies of GA-deficient plants. The genetic interaction of genes lka and lkb is examined and the action of gene lkb on a le gene background determined.     

Role of ethylene in alleviation of cadmium-induced photosynthetic capacity inhibition by sulphur in mustard

Sulphur (S) assimilation leads to the formation of glutathione (GSH) and alleviation of cadmium (Cd) stress. GSH is synthesized from its immediate metabolite cysteine, which also serves as a metabolite for ethylene formation through S‐adenosyl methionine. To assess the role of ethylene in S‐induced alleviation of Cd stress on photosynthesis, the effects of S or ethephon (ethylene source) on GSH and ethylene were examined in mustard (Brassica juncea L. cv. Varuna). Sufficient‐S at 100 mg S kg^?1 soil alleviated Cd‐induced photosynthetic inhibition more than excess‐S (200 mg S kg^?1 soil) via ethylene by increased GSH. Under Cd stress, plants were less sensitive to ethylene, despite high ethylene evolution, and showed photosynthetic inhibition. Ethylene sensitivity of plants increased with ethephon or sufficient‐S, triggering the induction of an antioxidant system, and leading to increased photosynthesis even under Cd stress. The effects of ethephon and S under Cd stress were similar. The effects of S were reversed by ethylene biosynthesis inhibitor, aminoethoxyvinylglycine (AVG), suggesting that ethylene plays an important role in S‐induced alleviation of Cd stress on photosynthesis.     

Jasmonates are essential factors inducing gummosis in tulips: mode of action of jasmonates focusing on sugar metabolism

The purpose of this study was to know the mechanism of jasmonates to induce gummosis in tulip (Tulipa gesneriana L. cv. Apeldoorn) shoots, especially on the focus of sugar metabolism. Gummosis in the first internode of tulip plants was induced by the application of methyl jasmonate (JA-Me, 1% w/w in lanolin) and jasmonic acid (JA, 1% w/w in lanolin) 5 days after application and strongly stimulated by the simultaneous application of ethylene-releasing compound, ethephon (2-chloroethylphosphonic acid, 1% w/w in lanolin), although ethephon alone had little effect. JA-Me stimulated ethylene production of the first internodes of tulips, ethylene production increasing up to more than 5 times at day 1 and day 3 after the application. On the other hand, application of ethephon did not increase endogenous levels of jasmonates in tulip stems. Analysis of composition of tulip gums revealed that they were consisted of glucuronoarabinoxylan with an average molecular weight of ca. 700 kDa. JA-Me strongly decreased the total amount of soluble sugars in tulip stems even in 1 day after application, being ca. 50% of initial values 5 days after application, but ethephon did not. However, both JA-Me and ethephon had almost no effect on the neutral sugar compositions of soluble sugars mainly consisting of glucose, mannose and xylose in ratio of 20:2:1 and traces of arabinose. Both JA-Me and ethephon applied exogenously stimulated senescence of tulip shoots shown by the loss of chlorophyll. These results strongly suggest that the essential factor of gummosis in tulips is jasmonates affecting the sugar metabolism in tulip shoots. The mode of action of jasmonates to induce gummosis of tulip shoots is discussed in relation to ethylene production, sugar metabolism and senescence.     

A Major Gene Conferring Reduced Ethylene Sensitivity and Maleness in Cucurbita pepo

External treatment with ethylene had indicated earlier that this hormone is the main factor controlling sex determination in Cucurbita pepo. Up to now, however, there was no genetic evidence that supported the relationship between ethylene production, or perception, and sexual expression in this species. Here we demonstrate that the extreme male phenotype of the Vegetable Spaghetti (Veg) inbred line of C. pepo subspecies pepo is determined by a major gene that confers reduced ethylene sensitivity in plants. The production of female flowers in the Veg line is very delayed and reduced with respect to the contrasting Bolognese (Bog) line, ranging between 5 and 35% of female flowers per plant. This enhanced maleness trait segregates as a single gene in the F₂ and backcross (BC) generations, and co-segregates with a weak ethylene-insensitive phenotype in the F₂ population, suggesting that the gene responsible for the Veg phenotype could be the result of a mutation in a receptor or response gene for ethylene. Although the etiolated seedlings of the Veg line, and the most androecious plants in the F₂ generation, produce more ethylene than those of the contrasting line, they are less sensitive to this hormone, as indicated by a weaker triple response and a delayed abscission of ethylene-treated male flowers. Given that the sexual phenotype of F₂ plants is correlated with ethylene sensitivity, with the more sensitive plants producing the higher number of female flowers, our results demonstrate that the ethylene response is directly involved in the control of sex determination in C. pepo. It regulates the induction of female flower production, and therefore the extension of the initial phase of development in which the plant produces only male flowers, as well as the number of female flowers per plant.     

The role of ethylene in the regeneration of Helianthus annuus (sunflower) plants from callus

Hypocotyl-derived callus from the Helianthus annuus L. inbred line SS415B regenerated significantly more plants if the seedlings were grown in the light. The difference between light- and dark-grown seedlings was not correlated with differences in seedling ethylene production, but seemed to be due to a difference in sensitivity to ethylene at a specific time during seedling growth. Treating 3-day-old dark-grown seedlings with 10 μ M aminoethoxyvinylglycine (AVG) effectively inhibited ethylene production for at least 7 days. Hypocotyl callus derived from AVG-treated seedlings gave the same amount of regeneration as callus from light-grown seedlings. Promotion of regeneration by AVG was not seen unless the 3-day-old seedlings were grown for 4 additional days prior to culturing hypocotyl explants. The effects of AVG could be reversed by treatment with 1-aminocyclopropane-1-carboxylic acid (ACC) during these 4 days. After the 4 days, ACC was no longer effective.     

Ethylene: a regulator of root architectural responses to soil phosphorus availability

The involvement of ethylene in root architectural responses to phosphorus availability was investigated in common bean ( Phaseolus vulgaris L.) plants grown with sufficient and deficient phosphorus. Although phosphorus deficiency reduced root mass and lateral root number, main root length was unchanged by phosphorus treatment. This resulted in decreased lateral root density in phosphorus-deficient plants. The possible involvement of ethylene in growth responses to phosphorus deficiency was investigated by inhibiting endogenous ethylene production with amino-ethoxyvinylglycine (AVG) and aerating the root system with various concentrations of ethylene. Phosphorus deficiency doubled the root-to-shoot ratio, an effect which was suppressed by AVG and partially restored by exogenous ethylene. AVG increased lateral root density in phosphorus- deficient plants but reduced it in phosphorus-sufficient plants. These responses could be reversed by exogenous ethylene, suggesting ethylene involvement in the regulation of main root extension and lateral root spacing. Phosphorus-deficient roots produced twice as much ethylene per g dry matter as phosphorus-sufficient roots. Enhanced ethylene production and altered ethylene sensitivity in phosphorus-deficient plants may be responsible for root responses to phosphorus deficiency.     

Pithiness in plants: I. The effect of mechanical perturbation and the involvement of ethylene in petiole pithiness in celery

Mechanical perturbation (MP) applied to celery (Appium graveolens L. cv. Florida 683) leaf petioles or ethephon application to the plant did not induce thigmomorphogenesis (inhibition of elongation and increase in thickness of the petiole). However, the two treatments did cause the parenchyma breakdown which leads to pithiness or increased natural pithiness, mainly at the base of the petiole. Nevertheless, MP (but not ethephon) decreased the severity of drought-stress or GA3-induced pithiness. Although MP stimulates ethylene production, mainly at the middle part of the petiole, it seems that the protection by MP of the petiole may not be directly mediated by ethylene production. The exposure of the plant to drought stress brought about an increase in ethylene evolution. Upon reirrigating the plants, the first steps of pithiness were accompanied by a sharp decline in ethylene production. This decrease might be due to membrane disruption. The increase in ethylene production during drought stress may be one of the events which stimulate pithiness of the celery leaf petiole.     

Phenotypic plasticity of stem elongation in two ecotypes of Stellaria longipes: the role of ethylene and response to wind

Using two ecotypes of Stellaria longipes an alpine form with low plasticity and a prairie form with high plasticity, we investigated whether ethylene was involved in the response to wind stress and might be important in controlling plasticity of stem elongation. Stem growth inhibition was positively correlated with concentration of ethephon application and elevation in ambient ethylene in alpine ecotypes, whereas stem growth in prairie plants was stimulated by low ethephon concentrations. When treated with high AVG, the effects were reversed: alpine plant growth was promoted and prairie plant growth was inhibited. Prairie plants exhibited a daily rhythm in ethylene evolution which increased and peaked at 1500 h, and which was absent in alpine plants. Ethylene evolution did not change significantly during the first 2 weeks of growth in alpine plants, whereas ethylene in prairie plants increased significantly during periods of rapid stem elongation. Wind treatment inhibited growth in both ecotypes, but only alpine plants showed a recovery of growth to control levels when wind stressed plants were pretreated with STS. In addition, only alpine plants showed an increase in ethylene evolution in response to wind simulation, whereas prairie plant ethylene evolution did not deviate from rhythms observed in unstressed plants. We concluded that ethylene dwarfs stems in alpine S. longipes in response to wind stress. However, low levels of ethylene may stimulate growth in prairie ecotypes and act independently of wind stress intensity. The contrasting ability to synthesize and respond to ethylene can account for part of the difference in plasticity documented between the two ecotypes.     

Further studies of auxin and ACC induced feminization in the cucumber plant using ethylene inhibitors

The present study was designed to establish the role of an essential hormone controlling sex expression in cucumber. A potent anti-ethylene agent, AgNO3, completely inhibited pistillate flower formation caused by IAA, ACC or ethephon. Inhibitors of ethylene biosynthesis, AVG and CoCl2 also suppressed feminization due to exogenous IAA or ACC. Though AVG also suppressed ethephon-induced feminization, this may be due to the second effect of AVG rather than the effect on ACC biosynthesis. These results confirm that ethylene is a major factor regulating feminization and that exogenous auxin induces pistillate flower formation through its stimulation of ethylene production, rather than ACC production.     

The role of ethylene and brassinosteroids in the control of sex expression and flower development in <Emphasis Type="Italic">Cucurbita pepo</Emphasis>

In this paper we compare the sensitivity of different squash genotypes to ethylene and brassinosteroids by studying the effects of different ethylene and brassinosteroid treatments on the sexual expression and flower development of different C. pepo genotypes: Bolognese (Bog) and Vegetable Spaghetti (Veg), two contrasting lines for ethylene production and sensitivity, as well as Cora, a standard commercial hybrid. Results have demonstrated that ethylene has a much greater effect on sexual expression and flower development in C. pepo than brassinosteroids. Ethephon increases the number of female flowers per plant and reduces the first male phase of development, while treatments with the ethylene inhibitors AVG and STS reduce the number of female flowers per plant and expand the first male phase of development. The differential response observed between genotypes appears to be related to ethylene production and sensitivity. Bog, which produces more ethylene and is more sensitive to this hormone, responded much better to AVG and STS, reducing the number of female flowers per plant, while Veg, which is characterised by lower production of and sensitivity to ethylene, responded better to ethephon by reducing the first male phase of development and increasing the number of female flowers per plant. The differential abortion of female and male flowers in ethephon, AVG and STS treatments, as well as the occurrence of bisexual flowers in the AVG and STS treated plants of the more ethylene sensitive genotypes, demonstrate that ethylene is also involved in the development of female flowers. Female flower buds require a minimal level of ethylene not only to complete their development and maturation without a premature abortion, but also to arrest the development of stamens in the third whorl and to promote the appropriate growth of the carpels. On the contrary, the role of brassinosteroids in the sexual expression of C. pepo was not so evident. The application of brassinazole, an inhibitor of brassinosteroid biosynthesis slightly changes the production of ethylene in the three analysed genotypes, but those changes have little effect on their sexual phenotypes, and they do not alter the development of the unisexual flowers.     

Stimulation and Inhibition by Ethephon of Stem and Leaf Growth of Some Gramineae at Different Stages of Development

The effect of ethephon on the growth of leaves and stems ofP. pratensis, Agrostis sp., and A. sativa was investigated.The length of the leaves of all species as well as that of theinternodes of stems of vegetative tillers of Agrostis sp. andof stems of reproductive A. sativa plants was reduced. Epidermalcell length was unchanged or somewhat increased, indicatinga reduction in cell number. The rate of cell elongation maybe decreased, however. A reduction in cell length was observedin the internode of generative oat plants that just startedto elongate at the time of treatment. In vegetative tillers of P.pratensis and A. sativa ethephonpromoted growth of the internodes by increasing cell length.Cell number was little affected. GA3 also promoted internodegrowth, causing an increase in cell length as well as in cellnumber. Application of ethephon and GA3 together gave an additiveeffect on cell length but a reverse effect on cell number. Effects of ethephon were observed for 5 or more weeks aftertreatment. Ethylene production from plants of A sativa increased,but the extra production had already decreased by 90% after11 d. No stimulation of the ethylene production by the plantsother than from the breakdown of ethephon was observed.     

Internode length in Pisum. Gibberellins and the slender phenotype

Pea plants ( Pisum sativum L.) possessing the slender phenotype (conferred by the gene combination la cry^s ) have extremely long, thin internodes and are phenotypically similar to dwarf plants (possessing genes La and/or Cry ) that have been treated with a non-limiting dose of gibberellin (GA₃). In contrast to tall and dwarf plants, slender plants are virtually insensitive to treatment with AMO 1618, PP333 or GA₃ and addition of the "gibberellin-less" mutant gene na does not alter the phenotype of slender plants. Na slender segregates possessed lower levels of gibberellin-like substances than comparable dwarf segregates when extracts from shoots were assayed using the lettuce hypocotyl or rice seedling bioassays. In addition, na slenders possessed little or no gibberellin-like activity even though they possessed a slender phenotype. Thus the gene combination la cry^s causes slender plants to respond as if they are saturated with gibberellins for growth. In addition, the gene combinations la cry^s and le la cry^c (allele cry^c is less extreme in effect than cry^s ) are shown to be almost completely epistatic to the alleles at the na locus. All these results suggest that gibberellin levels are not important in determining the internode length of slender peas (genotype la cry^s ). The possible mechanisms by which this could occur are discussed.     

Thigmomorphogenesis: Evidence for a translocatable thigmomorphogenetic factor induced by mechanical perturbation of beans (Phaseolus vulgaris)

Mechanical perturbation of bean (Phaseolus vulgaris L.) internodes results in reduced elongation and increased diameter of the internodes (thigmomorphogenesis). Perturbation of a single lower internode results in thigmorphogenesis in that internode and all of those internodes above it, the degree of which depends on the age (size) of the internodes and the frequency of perturbation. Application of ethephon to the internodes mimics mechanical perturbation. Early removal of the shoot tip or the cotyledons does not effect thigmomorphogenesis, indicating that those organs do not exert control over the response. Mechanical perturbation of one plant of a pair grafted together at the first internodes results in thigmomorphogenesis in both plants. This indicates the transport of some factor from the mechanically perturbed donor to the non-treated receiver. Evidence is presented to support the contention that ethylene is not this transportable factor.     

Ethylene reverses photosynthetic inhibition by nickel and zinc in mustard through changes in PS II activity,photosynthetic nitrogen use efficiency,and antioxidant metabolism

We investigated the influence of exogenously sourced ethylene (200 μL L^?1 ethephon) in the protection of photosynthesis against 200 mg kg^?1 soil each of nickel (Ni)- and zinc (Zn)-accrued stress in mustard (Brassica juncea L.). Plants grown with Ni or Zn but without ethephon exhibited increased activity of 1-aminocyclopropane carboxylic acid synthase, and ethylene with increased oxidative stress measured as H₂O₂ content and lipid peroxidation compared with control plants. The oxidative stress in Ni-grown plants was higher than Zn-grown plants. Under metal stress, ethylene protected photosynthetic potential by efficient PS II activity and through increased activity of ribulose-1,5-bisphosphate carboxylase and photosynthetic nitrogen use efficiency (P-NUE). Application of 200 μL L^?1 ethephon to Ni- or Zn-grown plants significantly alleviated toxicity and reduced the oxidative stress to a greater extent together with the improved net photosynthesis due to induced activity of ascorbate peroxidase and glutathione (GSH) reductase, resulting in increased production of reduced GSH. Ethylene formation resulting from ethephon application alleviated Ni and Zn stress by reducing oxidative stress caused by stress ethylene production and maintained increased GSH pool. The involvement of ethylene in reversal of photosynthetic inhibition by Ni and Zn stress was related to the changes in PS II activity, P-NUE, and antioxidant capacity was confirmed using ethylene action inhibitor, norbornadiene.