Increased Ethylene Sensitivity Research Articles

Evidence of the functional role of the ethylene receptor genes SlETR4 and SlETR5 in ethylene signal transduction in tomato.

Ethylene receptors are key factors for ethylene signal transduction. In tomato, six ethylene receptor genes (SlETR1-SlETR6) have been identified. Mutations in different ethylene receptor genes result in different phenotypes that are useful for elucidating the roles of each gene. In this study, we screened mutants of two ethylene receptor genes, SLETR4 and SLETR5, from a Micro-Tom mutant library generated by TILLING. We identified two ethylene receptor mutants with altered phenotypes and named them Sletr4-1 and Sletr5-1. Sletr4-1 has a mutation between the transmembrane and GAF domains, while Sletr5-1 has a mutation within the GAF domain. Sletr4-1 showed increased hypocotyl and root lengths, compared to those of wild type plants, under ethylene exposure. Moreover, the fruit shelf life of this mutant was extended, titratable acidity was increased and total soluble solids were decreased, suggesting a reduced ethylene sensitivity. In contrast, in the absence of exogenous ethylene, the hypocotyl and root lengths of Sletr5-1 were shorter than those of the wild type, and the fruit shelf life was shorter, suggesting that these mutants have increased ethylene sensitivity. Gene expression analysis showed that SlNR was up-regulated in the Sletr5-1 mutant line, in contrast to the down-regulation observed in the Sletr4-1 mutant line, while the down-regulation of SlCTR1, SlEIN2, SlEIL1, SlEIL3, and SlERF.E4 was observed in Sletr4-1 mutant allele, suggesting that these two ethylene receptors have functional roles in ethylene signalling and demonstrating, for the first time, a function of the GAF domain of ethylene receptors. These results suggest that the Sletr4-1 and Sletr5-1 mutants are useful for elucidating the complex mechanisms of ethylene signalling through the analysis of ethylene receptors in tomato.

Light, Ethylene and Auxin Signaling Interaction Regulates Carotenoid Biosynthesis During Tomato Fruit Ripening.

Light signaling and plant hormones, particularly ethylene and auxins, have been identified as important regulators of carotenoid biosynthesis during tomato fruit ripening. However, whether and how the light and hormonal signaling cascades crosstalk to control this metabolic route remain poorly elucidated. Here, the potential involvement of ethylene and auxins in the light-mediated regulation of tomato fruit carotenogenesis was investigated by comparing the impacts of light treatments and the light-hyperresponsive high pigment-2 (hp2) mutation on both carotenoid synthesis and hormonal signaling. Under either light or dark conditions, the overaccumulation of carotenoids in hp2 ripening fruits was associated with disturbed ethylene production, increased expression of genes encoding master regulators of ripening and higher ethylene sensitivity and signaling output. The increased ethylene sensitivity observed in hp2 fruits was associated with the differential expression of genes encoding ethylene receptors and downstream signaling transduction elements, including the downregulation of the transcription factor ETHYLENE RESPONSE FACTOR.E4, a repressor of carotenoid synthesis. Accordingly, treatments with exogenous ethylene promoted carotenoid biosynthetic genes more intensively in hp2 than in wild-type fruits. Moreover, the loss of HP2 function drastically altered auxin signaling in tomato fruits, resulting in higher activation of the auxin-responsive promoter DR5, severe down-regulation of AUXIN/INDOLE-3-ACETIC ACID (Aux/IAA) genes and altered accumulation of AUXIN RESPONSE FACTOR (ARF) transcripts. Both tomato ARF2 paralogues (Sl-ARF2a and SlARF2b) were up-regulated in hp2 fruits, which agrees with the promotive roles played by these ARFs in tomato fruit ripening and carotenoid biosynthesis. Among the genes differentially expressed in hp2 fruits, the additive effect of light treatment and loss of HP2 function was particularly evident for those encoding carotenoid biosynthetic enzymes, ethylene-related transcription factors, Aux/IAAs and ARFs. Altogether, the data uncover the involvement of ethylene and auxin as part of the light signaling cascades controlling tomato fruit metabolism and provide a new link between light signaling, plant hormone sensitivity and carotenoid metabolism in ripening fruits.

Identification and expression analysis of ethylene biosynthesis and signaling genes provides insights into the early and late coffee cultivars ripening pathway

The plant hormone ethylene is involved in the regulation of a multitude of plant processes, ranging from seed germination to organ senescence. Ethylene induces fruit ripening in climacteric fruits, such as coffee, being directly involved in fruit ripening time and synchronization. Coffee early cultivars usually show a more uniform ripening process although little is known about the genetic factors that promote the earliness of ripening. Thus, this work aimed to characterize the putative members of the coffee (Coffea arabica) ethylene biosynthesis and signaling pathways, as well as to analyze the expression patterns of these members during fruit ripening of early (Catucaí 785-15) and late (Acauã) coffee cultivars. Reverse Transcription-qPCR analysis of the four biosynthesis genes (CaACS1-like; CaACO1-like; CaACO4-like e CaACO5-like) analyzed in this study showed that CaACO1-like and CaACO4-like displayed an expression pattern typically observed in climacteric fruits, being up-regulated during ripening. CaACS1-like gene expression was also up-regulated during fruit ripening of both cultivars, although in a much lesser extent when compared to the changes in CaACO1-like and CaACO4-like gene expression. CaACO5-like was only induced in raisin fruit and may be related to senescence processes. On the other hand, members of the ethylene signaling pathway (CaETR1-like, CaETR4-like, CaCTR2-like, CaEIN2-like, CaEIN3-like, CaERF1) showed slightly higher expression levels during the initial stages of development (green and yellow-green fruits), except for the ethylene receptors CaETR1-like and CaETR4-like, which were constitutively expressed and induced in cherry fruits, respectively. The higher ethylene production levels in Catucaí 785-15 fruits, indicated by the expression analysis of CaACO1-like and CaACO4-like, suggest that it promotes an enhanced CaETR4-like degradation, leading to an increase in ethylene sensitivity and consequently to an earliness in the ripening process of this cultivar. Ethylene production in Acauã fruits may not be sufficient to inactivate the CaETR4-like levels and thus ripening changes occur in a slower pace. Thus, the expression analysis of the ethylene biosynthesis and signaling genes suggests that ethylene is directly involved in the determination of the ripening time of coffee fruits, and CaACO1-like, CaACO4-like and CaETR4-like may display essential roles during coffee fruit ripening.

The AP2/ERF Transcription Factor AtERF73/HRE1 Modulates Ethylene Responses during Hypoxia in Arabidopsis

A number of APETALA2 (AP2)/ETHYLENE RESPONSE FACTOR (ERF) genes have been shown to function in abiotic and biotic stress responses, and these genes are often induced by multiple stresses. We report here the characterization of an AP2/ERF gene in Arabidopsis (Arabidopsis thaliana) that is specifically induced during hypoxia. We show that under normoxic conditions, the expression of AtERF73/HRE1 can be induced by exogenous addition of 1-aminocyclopropane-1-carboxylic acid and that a combination of hypoxia and 1-aminocyclopropane-1-carboxylic acid results in hyperinduction of AtERF73/HRE1 expression. In addition, hypoxic induction of AtERF73/HRE1 is reduced but not completely abolished in ethylene-insensitive mutants and in the presence of inhibitors of ethylene biosynthesis and responses. These results suggest that, in addition to ethylene, an ethylene-independent signal is also required to mediate hypoxic induction of AtERF73/HRE1. To assess the role of AtERF73/HRE1, we generated three independent RNA interference (RNAi) knockdown lines of AtERF73/HRE1. Under normoxic conditions, the AtERF73/HRE1-RNAi seedlings displayed increased ethylene sensitivity and exaggerated triple responses, indicating that AtERF73/HRE1 might play a negative regulatory role in modulating ethylene responses. Gas chromatography analyses showed that the production of ethylene was similar between wild-type and RNAi lines under hypoxia. Quantitative reverse transcription-polymerase chain reaction analyses showed that hypoxia-inducible genes could be affected by AtERF73/HRE1-RNAi lines in two different ways: hypoxic induction of glycolytic and fermentative genes was reduced, whereas induction of a number of peroxidase and cytochrome P450 genes was increased. Taken together, our results show that AtERF73/HRE1 is involved in modulating ethylene responses under both normoxia and hypoxia.

Endogenous auxin regulates the sensitivity of Dendrobium (cv. Miss Teen) flower pedicel abscission to ethylene.

Dendrobium flower buds and flowers have an abscission zone at the base of the pedicel (flower stalk). Ethylene treatment of cv. Miss Teen inflorescences induced high rates of abscission in flower buds but did not affect abscission once the flowers had opened. It is not known if auxin is a regulator of the abscission of floral buds and open flowers. The hypotheses that auxin is such a regulator and is responsible for the decrease in ethylene sensitivity were tested. Severed inflorescences bearing 4-8 floral buds and 4-6 open flowers were used in all tests. The auxin antagonists 2,3,5-triiodobenzoic acid (TIBA, an inhibitor of auxin transport) or 2-(4-chlorophenoxy)-2-methyl propionic acid (CMPA, an inhibitor of auxin action) were applied to the stigma of open flowers. Both chemicals induced high flower abscission rates, even if the inflorescences were not treated with ethylene. The effects of these auxin antagonists virtually disappeared when the inflorescences were treated with 1-methylcyclopropene (1-MCP), indicating that the abscission induced by the auxin antagonists was due to ethylene. Removal of the open flowers at the distal end of the pedicel hastened the time to abscission of the remaining pedicel, and also resulted in an increase in ethylene sensitivity. Indole-3-acetic acid (IAA) in lanolin, placed on the cut surface of the pedicel, replaced the effect of the removed flower. Treatments that promoted abscission of open flowers up-regulated a gene encoding a β-1,4-glucanase (Den-Cel1) in the abscission zone (AZ). The abundance of Den-Cel1 mRNA was highly correlated with β-1,4-glucanase activity in the AZ. The results show that auxin is an endogenous regulator of floral bud and flower abscission and suggest that auxin might explain, at least partially, why pedicel abscission of Dendrobium cv. Miss Teen changes from very ethylene-sensitive to ethylene-insensitive.

Sl-ERF2, a Tomato Ethylene Response Factor Involved in Ethylene Response and Seed Germination

Ethylene response factors (ERFs) are plant transcriptional regulators mediating ethylene-dependent gene expression via binding to the GCC motif found in the promoter region of ethylene-regulated genes. We report here on the structural and functional characterization of the tomato Sl-ERF2 gene that belongs to a distinct class of the large ERF gene family. Both spliced and unspliced versions of Sl-ERF2 transcripts were amplified from RNA samples and the search in the public tomato expressed sequence tag (EST) database confirmed the existence of the two transcript species in a number of cDNA libraries. The unspliced transcript contains two open reading frames yielding two hypothetical proteins, a small highly truncated version lacking the APETALA2 domain and a bigger protein lacking the N-terminal MCGGAAI(I)/(L) consensus peptide specific to ERF members from subfamily IV. Nevertheless, functional Sl-ERF2 protein may only derive from spliced transcripts since, depending on the tissue, the level of the spliced transcript is much higher than that of the unspliced transcript. Sl-ERF2 is expressed in all plant tissues tested, though its transcript accumulates preferentially in germinating seeds and ripening fruit. Overexpression of the Sl-ERF2 gene in transgenic tomato lines results in premature seed germination and enhanced hook formation of dark-grown seedlings, which is indicative of increased ethylene sensitivity. The expression of the mannanase2 gene is upregulated in Sl-ERF2-overexpressing seeds, suggesting that Sl-ERF2 stimulates seed germination through the induction of the mannanase2 gene. It is noteworthy that the exaggerated hook phenotype is abolished when ethylene perception is blocked, strongly suggesting that Sl-ERF2 requires other ethylene-dependent components to impact the hook formation process.

Stem and leaf growth of alpine sun and prairie shade ecotypes of Stellaria longipes under different photoperiods: role of ethylene

Plant ecotypes of Stellaria longipes Goldie from competitive, shade-adapted prairie habitat and less competitive, nonshaded alpine habitat were subjected to shortened or extended photoperiod conditions. Increasing daylength was positively correlated to increased stem elongation in both ecotypes. Leaf length and area for shade (prairie) plants was significantly altered with increased photoperiods, whereas sun (alpine) plants exhibited minimal leaf expansion in response to increased photoperiod. Increased ethylene evolution in the alpine genotype during rapid stem elongation and extended photoperiods suggests that ethylene plays a growth regulatory role in this sun-adapted genotype. The prairie genotype evolved less ethylene during these same periods, indicating either a diminished requirement for elevated ethylene to effect elongation and leaf expansion responses or possibly increased ethylene sensitivity because of interactions with other hormones, such as gibberellin or auxin. The sun genotype consistently produced more ethylene than the shade genotype under all photoperiod treatments. We conclude that photoperiod alters stem elongation and leaf expansion responses; similar trends were observed for extended photoperiods as were observed for shaded conditions, specifically low light intensity (photosynthetically active radiation) conditions. Further, ethylene levels altered during these responses, especially in sun-adapted alpine plants, which suggests that ethylene is involved in these growth responses.

High floral bud abscission and lack of open flower abscission in Dendrobium cv. Miss Teen: rapid reduction of ethylene sensitivity in the abscission zone.

We studied the abscission of floral buds and open flowers in cut Dendrobium inflorescences. Abscission of floral buds was high and sensitive to ethylene in all cultivars studied. Many open flowers abscised in most cultivars, but cv. Willie exhibited only small amount of floral fall and cv. Miss Teen none. Applied ethylene (0.4 μL L-1 for 24 h at 27°C) greatly hastened abscission of open flowers in most cultivars, but had only a small effect in cv. Willie and no effect in cv. Miss Teen. Flower fall, if it occurred, was completely inhibited by 1-methylcyclopropene (1-MCP), showing that it was regulated by endogenous ethylene. Ethylene production from the abscission zones was low in all cultivars studied. In cv. Miss Teen the abscission zone changed from highly ethylene sensitive to completely insensitive in ~30 h, coinciding with floral opening. Removal of the floral buds somewhat reduced abscission in open flowers, but the lack of open flower abscission in cv. Miss Teen could not be explained by higher bud fall. The ovary did not grow in the (unpollinated) flowers, showing that lack of abscission in cvv. Willie and Miss Teen was not due to parthenocarpy. Flower removal in cv. Miss Teen had no effect on ethylene sensitivity of the abscission of the remaining pedicel. However, removal of the distal 2 cm of the 3-cm-long pedicels dramatically increased ethylene sensitivity. This suggests that the pedicel is important for the low ethylene insensitivity of abscission, in this cultivar. It is concluded that the abscission zones in the cvv. Willie and Miss Teen, in contrast with the other cultivars investigated, became rapidly insensitive to ethylene at the time of flower opening. At least part of the ethylene sensitivity in Miss Teen seems to be due to a factor in the pedicel.

Mutation of a UDP‐glucose‐4‐epimerase alters nematode susceptibility and ethylene responses in Arabidopsis roots

In Arabidopsis, mutation of RHD1, a UDP-glucose-4-epimerase, causes root-specific phenotypes, including hypersusceptibility to the cyst nematode Heterodera schachtii, increased root hair elongation, decreased root length, and root epidermal bulging. Previous experiments suggested that increased ethylene sensitivity or production mediated the rhd1-4 phenotypes. In the present study, double mutant analyses revealed that only rhd1-4 hypersusceptibility to H. schachtii and increased root hair elongation were dependent upon the ethylene signaling genes EIN2 and EIN3 but not upon ethylene signaling mediated by the auxin efflux carrier EIR1. In contrast, the rhd1-4 short root and root epidermal bulging phenotypes did not require EIN2, EIN3, or EIR1. A time-course analysis of RHD1 transcript levels in wild-type plants treated with the ethylene precursor 1-aminocyclopropane-1-carboxylic acid showed a root-specific downregulation of RHD1 expression by ethylene. This observation was corroborated by our finding of increased RHD1 transcript levels in roots of the ethylene-insensitive mutants etr1 and ein2. In addition to ethylene, auxin strongly influences H. schachtii susceptibility and root hair elongation. Therefore, we investigated the sensitivity of rhd1-4 roots to indole-3-acetic acid (IAA). Equivalent IAA concentrations caused a greater reduction in rhd1-4 root elongation compared with wild-type roots. Finally, H. schachtii parasitism was found to strongly downregulate RHD1 expression in the root 3 days after inoculation. We conclude that RHD1 is a likely target of root-specific negative regulation by ethylene and that loss of RHD1 function results in a heightened sensitivity of root tissues to both ethylene and auxin.

Leaf Yellowing of Cut Standard Chrysanthemum (Dendranthema grandiflora Kitamura) 'Shuho-no-chikara' Induced by Ethylene and the Postharvest Increase in Ethylene Sensitivity

The sensitivity of chrysanthemum (Dendranthema grandiflora Kitamura) 'Shuho-no-chikara' to ethylene, which causes postharvest leaf yellowing (senescence), was examined. A 24-hr exposure to 1000 ppm ethylene on day 1 or 3 after harvest did not induce leaf yellowing throughout 20-day postharvest period, although the same treatment on day 4 or 6 did. Exposure as low as 1 ppm for 24 hr to ethylene on day 10 induced leaf yellowing. When cut flowers were exposed to ethylene on day 5, the effect depended on the duration of exposure and concentration; a 12- or 24-hr exposure at 1 ppm did not induce leaf yellowing, whereas a 48-hr exposure or a 24-hr exposure at 1000 ppm did. With potted flowering plants, exposure to 1000 ppm ethylene for 24 hr on day 1, 5 or 10 did not induce yellowing of leaves under the same postharvest-environmental conditions as cut flowers. These results indicate that exogenous ethylene promotes leaf yellowing in cut 'Shuho-no-chikara' chrysanthemums and that sensitivity of leaves to ethylene gradually increases with time. In continuously ethylene-treated and untreated leaves of cut vegetative shoots, ethylene production rate peaked 3 and 5 days after harvest, respectively, and then declined. The respiration rate was slightly higher in ethylene-treated leaves. The level of 1-aminocyclopropane-1-carboxylic acid (ACC) content began to increase several days after harvest. The level was higher in continuously ethylene-treated leaves. The relationship between these parameters and the sensitivity to ethylene are discussed.

Effect of octanoic acid on ethylene-mediated processes in Arabidopsis

We have examined whether octanoic acid (OA) one of the short chain saturated fatty acids (SCSFA), increases ethylene response in the following three ethylene-mediated processes: a) hypocotyl growth in darkness; b) formation of new flowers; c) flower abscission. These processes were examined in the presence or absence of exogenous ethylene in Arabidopsis wild type (WT) and in the ethylene-insensitive mutants, etr1-3 and ein2-1 and in the ethylene over-producer mutant eto1-1. Our results show that OA decreased hypocotyl length of WT in the absence or presence of exogenous ethylene, apparently showing that OA acts via augmentation of ethylene action. However, the hypocotyl growth inhibition could not be ascribed to increased ethylene sensitivity since application of inhibitors of ethylene synthesis (aminoethoxyvinylglycine; AVG) or action (1-methylcyclopropene;1-MCP) to WT seedlings did not prevent specifically the OA-induced growth inhibition. Also, OA inhibited hypocotyl growth in the mutants etr1-3 and ein2-1 in a similar pattern to that obtained in WT. On the other hand, OA had no effect on flower formation neither in WT, etr1-3 and eto1-1, in which ethylene reduced flower formation, nor in the ein2-1 mutant, in which ethylene had no effect. OA also did not increase flower abscission in WT or in the mutants etr1-3 and ein2-1 neither in the absence nor in the presence of ethylene. However, OA has augmented flower abscission in the mutant eto1-1 only in the absence of exogenous ethylene. This result might indicate that the effect of OA on eto1-1 is specific to this mutant and is not due to general deleterious effects inflicted by OA. Taken together, our results show that in general OA does not augment ethylene response in Arabidopsis, but it might affect ethylene action in flower abscission of the ethylene-overproducer mutant.

Reduced expression of the tomato ethylene receptor gene LeETR4 enhances the hypersensitive response to Xanthomonas campestris pv. vesicatoria.

The hypersensitive response (HR) involves rapid death of cells at the site of pathogen infection and is thought to limit pathogen growth through the plant. Ethylene regulates senescence and developmental programmed cell death, but its role in hypersensitive cell death is less clear. Expression of two ethylene receptor genes, NR and LeETR4, is induced in tomato (Lycopersicon esculentum cv. Mill) leaves during an HR to Xanthomonas campestris pv. vesicatoria, with the greatest increase observed in LeETR4. LeETR4 antisense plants previously were shown to exhibit increased sensitivity to ethylene. These plants also exhibit greatly reduced induction of LeETR4 expression during infection and an accelerated HR at inoculum concentrations ranging from 10(5) to 10(7) CFU/ml. Increases in ethylene synthesis and pathogenesis-related gene expression are greater and more rapid in infected LeETR4 antisense plants, indicating an enhanced defense response. Populations of avirulent X. campestris pv. vesicatoria decrease more quickly and to a lower level in the transgenic plants, indicating a greater resistance to this pathogen. Because the ethylene action inhibitor 1-methylcyclopropene alleviates the enhanced HR phenotype in LeETR4 antisense plants, these changes in pathogen response are a result of increased ethylene sensitivity.

The Arabidopsis eer1 mutant has enhanced ethylene responses in the hypocotyl and stem.

By screening for enhanced ethylene-response (eer) mutants in Arabidopsis, we isolated a novel recessive mutant, eer1, which displays increased ethylene sensitivity in the hypocotyl and stem. Dark-grown eer1 seedlings have short and thick hypocotyls even in the absence of added ethylene. This phenotype is suppressed, however, by the ethylene biosynthesis inhibitor 1-aminoethoxyvinyl-glycine. Following ethylene treatment, the dark-grown eer1 hypocotyl response is greatly exaggerated in comparison with the wild type, indicating that the eer1 phenotype is not simply due to ethylene overproduction. eer1 seedlings have significantly elevated levels of basic-chitinase expression, suggesting that eer1 may be highly sensitive to low levels of endogenous ethylene. Adult eer1 plants display exaggerated ethylene-dependent stem thickening, which is an ethylene response previously unreported in Arabidopsis. eer1 also has enhanced responsiveness to the ethylene agonists propylene and 2,5-norbornadiene. The eer1 phenotype is completely suppressed by the ethylene-insensitive mutation etr1-1, and is additive with the constitutive ethylene-response mutation ctr1-3. Our findings suggest that the wild-type EER1 product acts to oppose ethylene responses in the hypocotyl and stem.

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.

Short‐chain saturated fatty acids in the regulation of pollination‐induced ethylene sensitivity of Phalaenopsis flowers

Pollination greatly accelerates petal senescence. The first observed event after pollination is an increase in the flower's sensitivity to ethylene, followed by an increase in ethylene biosynthesis. Our objectives were to study the mode of action of the increase in ethylene sensitivity and the possible involvement of short‐chain saturated fatty acids (SCSFAs) in this process. Application of SCSFAs, ranging in chain length from 7 to 10 carbons onto stigmas of Phalaenopsis (Phalaenopsis hybrid, cv. Herbert Hager) flowers increased their sensitivity to ethylene in the same way as pollination. Following pollination, there was a significant increase in the endogenous content of these fatty acids in the flower's column and perianth, with octanoic acid (C8) being the main SCSFA observed. The increase in SCSFA content was observed as early as 6 h after pollination and began to decline 6 h later. Incorporation of octanoic acid into liposomes or microsomal membranes isolated from Phalaenopsis petals resulted in a decrease in lipid order that was detected by fluorescence polarization of dansyl pyrrolidine (DNSP) but not of 1,6‐diphenyl‐1,3,5‐hexatriene (DPH). At peak ethylene sensitivity, 10 h after pollination, there was a significant decrease in the lipid order of microsomal membranes isolated from Phalaenopsis columns and perianths, again as detected by DNSP but not by DPH. Stigmatic application of octanoic acid mimicked the effect of pollination on membrane lipid order. We suggest that SCSFAs may be the ethylene ‘sensitivity factors’ produced following pollination, and that their mode of action involves a decrease in the order of specific regions in the membrane lipid bilayer, consequently altering ethylene action.

Promotive effect of jasmonates on the senescence of detached maize leaves

Promotion of senescence of detached maize leaves by jasmonates was investigated. Senescence of detached maize leaves was promoted by linolenic acid, the precursor of biosynthesis of jasmonic acid, and retarded by inhibitors of lipoxygenase, the first enzyme in the biosynthetic pathway of jasmonic acid. Results support a role of endogenous jasmonates in the regulation of senescence of detached maize leaves. Silver thiosulfate, an inhibitor of ethylene action, was found to inhibit methyl jasmonate, linolenic acid- and abscisic acid-promoted senescence of detached maize leaves. It seems that jasmonate-promoted senescence is mediated through an increase in ethylene sensitivity in detached maize leaves.

An increase in ethylene sensitivity following pollination is the initial event triggering an increase in ethylene production and enhanced senescence of Phalaenopsis orchid flowers

Cut Phalaenopsis (Phalaenopsis hybrid, cv. Herbert Hager) flowers usually last about 2 weeks. Following pollination however, there is a rapid acceleration of the wilting process, which is completed within 2 days. The first event detected following pollination was an increase in ethylene sensitivity. This increased sensitivity began about 4 h after pollination and peaked 6 h later. A subsequent increase in ethylene production could only be detected 12 to 14 h after pollination. Treatment of the flowers with silver thiosulfate or 1‐methylcyclopropene, both inhibitors of ethylene action, completely inhibited the pollination‐induced increase in ethylene production and the enhanced senescence of the flowers. This indicates that the pollination‐induced increase in ethylene production is a response to the existing ethylene. Treatment of flowers with calcium and its ionophore A23187, which increased ethylene sensitivity and protein phosphorylation, also promoted ethylene production and senescence of unpollinated flowers, EGTA, a calcium chelator, decreased the sensitivity of pollinated flowers to ethylene and delayed and decreased the pollination‐induced increase in ethylene production. We suggest that the pollination‐induced increase in ethylene sensitivity is the initial pollination‐induced event triggering the enhancement of ethylene production, which leads to enhanced senescence of Phalaenopsis flowers.

An increase in ethylene sensitivity following pollination is the initial event triggering an increase in ethylene production and enhanced senescence of Phalaenopsis orchid flowers

An increase in ethylene sensitivity following pollination is the initial event triggering an increase in ethylene production and enhanced senescence of Phalaenopsis orchid flowers

Role of Ethylene and Short-Chain Saturated Fatty Acids in the Smoke-Stimulated Germination of Cyclopia Seed

Summary The seeds of both Cyclopia intermedia and Cyclopia subternata exhibited a seed coat imposed dormancy. However, seeds of the former species also exhibited an embryo dormancy which could be broken by exposure to smoke or ethylene. This stimulating effect of smoke and ethylene was inhibited by exposure to NBD, indicating that ethylene in the smoke was responsible for the stimulation of germination. The involvement of ethylene in germination in both species was clearly demonstrated by the inhibition of germination after treatment with AOA. Differences in germination between the two species can not be ascribed to differences in ethylene production. Seeds of C. subternata , however, showed a much higher capacity to bind ethylene than seeds of C. intermedia . Pulse treatments with octanoic acid or smoke-saturated water stimulated germination in C. intermedia due to an increase in ethylene sensitivity, while longer treatments inhibited germination in both species. It appears that octanoic acid present in smokesaturated water was responsible for the reaction of the seeds to the water.

Pollination-induced changes in ethylene production and sensitivity to ethylene in cut dendrobium orchid flowers

The longevity of cut dendrobium orchid (Dendrobium hybrid, cultivar ‘Jaquelyn Thomas’) flowers is normally 3 weeks. After pollination however, there is a rapid acceleration of the wilting process, beginning after only 24 h. Enhancement of senesence was induced only by successful pollination and not by emasculation. Following pollination there was an increase in both ethylene production and the sensitivity to ethylene. The increase in ethylene production began about 12 h after pollination and reached its peak after 30 h. An increase in ethylene sensitivity following pollination was not dependent on endogenous ethylene production as it occurred also in flowers treated with (aminooxy) acetic acid, an inhibitor of ethylene biosynthesis, which greatly reduced ethylene production levels.