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Abiotic stresses have a negative effect on crop production, affecting both vegetative and reproductive development. Ethylene plays a relevant role in plant response to environmental stresses, but the specific contribution of ethylene biosynthesis and signalling components in the salt stress response differs between Arabidopsis and rice, the two most studied model plants. In this paper, we study the effect of three gain-of-function mutations affecting the ethylene receptors CpETR1B, CpETR1A, and CpETR2B of Cucurbita pepo on salt stress response during germination, seedling establishment, and subsequent vegetative growth of plants. The mutations all reduced ethylene sensitivity, but enhanced salt tolerance, during both germination and vegetative growth, demonstrating that the three ethylene receptors play a positive role in salt tolerance. Under salt stress, etr1b, etr1a, and etr2b germinate earlier than WT, and the root and shoot growth rates of both seedlings and plants were less affected in mutant than in WT. The enhanced salt tolerance response of the etr2b plants was associated with a reduced accumulation of Na+ in shoots and leaves, as well as with a higher accumulation of compatible solutes, including proline and total carbohydrates, and antioxidant compounds, such as anthocyanin. Many membrane monovalent cation transporters, including Na+/H+ and K+/H+ exchangers (NHXs), K+ efflux antiporters (KEAs), high-affinity K+ transporters (HKTs), and K+ uptake transporters (KUPs) were also highly upregulated by salt in etr2b in comparison with WT. In aggregate, these data indicate that the enhanced salt tolerance of the mutant is led by the induction of genes that exclude Na+ in photosynthetic organs, while maintaining K+/Na+ homoeostasis and osmotic adjustment. If the salt response of etr mutants occurs via the ethylene signalling pathway, our data show that ethylene is a negative regulator of salt tolerance during germination and vegetative growth. Nevertheless, the higher upregulation of genes involved in Ca2+ signalling (CpCRCK2A and CpCRCK2B) and ABA biosynthesis (CpNCED3A and CpNCED3B) in etr2b leaves under salt stress likely indicates that the function of ethylene receptors in salt stress response in C. pepo can be mediated by Ca2+ and ABA signalling pathways. [ABSTRACT FROM AUTHOR]

Ethylene, a regulator of plant growth and development, is perceived by specific receptors that act as negative regulators of the ethylene response. Five ethylene receptors, i.e., ETR1, ERS1, EIN4, ETR2, and ERS2, are present in Arabidopsis and dominant negative mutants of each that confer ethylene insensitivity have been reported. In contrast, maize contains just two types of ethylene receptors: ZmERS1, encoded by ZmERS1a and ZmERS1b, and ZmETR2, encoded by ZmETR2a and ZmETR2b. In this study, we introduced a Cys to Tyr mutation in the transmembrane domain of ZmERS1b and ZmETR2b that is present in the etr1-1 dominant negative mutant and expressed each protein in Arabidopsis. Mutant Zmers1b and Zmetr2b receptors conferred ethylene insensitivity and Arabidopsis expressing Zmers1b or Zmetr2b were larger and exhibited a delay in leaf senescence characteristic of ethylene insensitive Arabidopsis mutants. Zmers1b and Zmetr2b were dominant and functioned equally well in a hemizygous or homozygous state. Expression of the Zmers1b N-terminal transmembrane domain was sufficient to exert dominance over endogenous Arabidopsis ethylene receptors whereas the Zmetr2b N-terminal domain failed to do so. Neither Zmers1b nor Zmetr2b functioned in the absence of subfamily 1 ethylene receptors, i.e., ETR1 and ERS1. These results suggest that Cys65 in maize ZmERS1b and ZmETR2b plays the same role that it does in Arabidopsis receptors. Moreover, the results demonstrate that the mutant maize ethylene receptors are functionally dependent on subfamily 1 ethylene receptors in Arabidopsis, indicating substantial functional conservation between maize and Arabidopsis ethylene receptors despite their sequence divergence.

The effects of ethylene and ethylene signal transduction have been long studied in plants. It is only recently reported that a single‐celled cyanobacterium, Synechocystis, contains a functional ethylene receptor. However, it remains to be determined if functional ethylene receptors are more widespread. This review summarizes what is known about responses to ethylene, ethylene receptors, and signal transduction in nonplant species. A summary is provided of information gleaned from searches of sequenced genomes and the possibility that ethylene receptors are widespread in microbes is discussed. [ABSTRACT FROM AUTHOR]

Key message: H2 prolonged the vase life and improved the vase quality of cut roses through repressing endogenous ethylene production and alleviating ethylene signal transduction during the entire senescing period. Recently, the application of hydrogen gas (H2) was shown to improve postharvest quality and longevity in perishable horticultural products, but the specific regulation mechanism remains obscure. Here, endogenous ethylene production and the expression of genes in ethylene biosynthesis and signalling pathway were investigated to explore the crosstalk between H2 and ethylene during the senescence of cut roses. Our results revealed that addition of exogenous ethylene by ethephon accelerated the senescence of cut roses, in which 100 mg L−1 ethephon displayed the most obvious senescent phenotype. While the applied different concentrations (1%, 10%, 50% and 100%) of hydrogen-rich water (HRW) conducted different affects in alleviating the senescence of cut roses, and 1% HRW displayed the best ornamental quality and the longest vase life by reducing ethylene production, supported by the decrease of 1-aminocyclopropene-1-carboxylate (ACC) accumulation, ACC synthase (ACS) and ACC oxidase (ACO) activities, and Rh-ACS3 and Rh-ACO1 expressions in ethylene biosynthesis. In addition, HRW increased the transcripts of ethylene receptor genes Rh-ETR1 at blooming period from day 4 to day 6 and suppressed Rh-ETR3 at senescence phase at day 8 after harvest. Furthermore, the relevant affection of HRW on Rh-ETR1 and Rh-ETR3 expressions still existed when the ethylene production was compromised by adequate addition of exogenous ethylene in HRW-treated cut rose petals, and HRW directly repressed the protein level of Rh-ETR3 in a transient expression assay. Overall, the results suggested that H2 is involved in neutralizing ethylene-mediated postharvest in cut flowers. [ABSTRACT FROM AUTHOR]

BACKGROUND Processes extending the shelf life of climacteric fruit play an important role in terms of a sustainable global food supply. In a previous study, a synthetic octapeptide (NOP‐1) was shown to inhibit the interaction between ethylene receptor (ETR) and ethylene insensitive‐2 (EIN2), and in consequence delay tomato ripening. We investigated for the first time the effect of NOP‐1 on inhibiting the ripening of apples ('Golden Delicious') during postharvest. RESULTS: Using purified recombinant proteins from a bacterial expression system, we demonstrate here that EIN2 also interacts tightly (Kd = 136 ± 29 nmol L−1) with the corresponding apple ETR MdETR1. In line with previous binding studies on tomato ETRs, the ripening‐delaying peptide NOP‐1 clearly binds to the purified apple ETR. An NOP‐1 solution (1000 µmol L−1) was applied with a brush or microdispenser and compared with apples treated with 1‐methylcyclopropene (1‐MCP) (SmartFresh™, Agrofresh) applied as gaseous treatment or untreated control fruits. NOP‐1 inhibited colour development and chlorophyll degradation during shelf life. These effects were more pronounced with the brush application (surface film) than with microdroplets application (mimicking a sprayable formulation). NOP‐1 did not alter ethylene release or respiration rate, whereas 1‐MCP expectedly strongly suppressed both. There were no differences in quality parameters evaluated. CONCLUSION: Our study shows that NOP‐1 binds to MdETR1 which results in delaying of ethylene‐dependent ripening developments of skin colour and chlorophyll. Besides application methods, possible reasons for the weak effect of NOP‐1 in comparison with previous tomato experiments could be different receptor affinity and penetration differences. © 2019 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

The effects of two oxylipins - jasmonic acid (JA) and 9, 10-ketol-octadecadienoic acid (KODA) - on ethylene metabolism and ethylene receptor genes were examined in peach fruit (Prunus persica L.) harvested at 88 days after full bloom (DAFB), in the pre-climacteric stage, and at 102 DAFB, at the beginning of the climacteric stage. Immediate post-harvest applications of either n-propyl dihydrojasmonate (PDJ) (a JA analog) or KODA stimulated ethylene production and 1-aminocyclopropane-1-carboxylate (ACC) concentrations in fruit that were harvested at 88 and 102 DAFB. KODA application significantly increased levels of ACC oxidase (PpACO1) expression at 5 and 8 days after treatment (DAT) in 88-DAFB-harvested fruit, and at 1 DAT in 102-DAFB-harvested fruit. Expression levels of ethylene response factors (PpERS1) were not influenced by either PDJ or KODA applications at 88 or 102 DAFB. In contrast, levels of ethylene-receptor-coding genes (PpETR1) expression in 88-DAFB KODA-treated fruit increased at 8 and 14 DAT. Expression levels of constitutive triple response (PpCTR1) in the untreated control fruit decreased with DAT in both 88- and 102-DAFB-harvested fruit. These results suggest that certain oxylipins may promote ethylene production in peach fruit, and that PpETR1 and PpCTR1 may be associated with the autocatalytic system of ethylene production. [ABSTRACT FROM AUTHOR]

The effects of paper containing different concentrations of 1‐methylcyclopropene (1‐MCP) on the quality of Chinese mushrooms were investigated. Mushrooms were first stored with paper containing 0, 0.25, 0.50, 0.75, 1.00, or 1.25 μl L−1 1‐MCP at 15 ± 1°C for 12 hr and then continued for up to 8 days. 1‐MCP reduced the respiratory peak, weight loss, degree of cap browning, relative leakage rate, and soluble solid content, and delayed the change in the surface hue angle. Mushrooms exposed to paper containing 1‐MCP maintained firmness and sensory attributes. The storage time of Chinese mushrooms treated with 1‐MCP at the best concentration (0.75 μl L−1) was 8 days, which was 4 days longer than that of the control. Browning in mushrooms was related to the vitamin C content, total phenol content, and polyphenol oxidase activity. The results would be very useful to the mushroom industry and provide a theoretical basis for the potential researches of the mechanisms maturation and senescence in Chinese mushrooms. The results in this article could provide a theoretical basis for the potential use of paper containing 1‐MCP in the development of Chinese mushroom commercial production. [ABSTRACT FROM AUTHOR]

Ethylene, the plant ripening hormone of climacteric fruit, is perceived by ethylene receptors which is the first step in the complex ethylene signal transduction pathway. Much progress has been made in elucidating the mechanism of this pathway, but there is still a lot to be done in the proteomic quantification of the main proteins involved, particularly during fruit ripening. This work focuses on the mass spectrometry based identification and quantification of the ethylene receptors (ETRs) and the downstream components of the pathway, CTR-like proteins (CTRs) and ETHYLENE INSENSITIVE 2 (EIN2). We used tomato as a model fruit to study changes in protein abundance involved in the ethylene signal transduction during fruit ripening. In order to detect and quantify these low abundant proteins located in the membrane of the endoplasmic reticulum, we developed a workflow comprising sample fractionation and MS analysis using parallel reaction monitoring. This work shows the feasibility of the identification and absolute quantification of all seven ethylene receptors, three out of four CTRs and EIN2 in four ripening stages of tomato. In parallel, gene expression was analyzed through real-time qPCR. Correlation between transcriptomic and proteomic profiles during ripening was only observed for three of the studied proteins, suggesting that the other signaling proteins are likely post-transcriptionally regulated. Based on our quantification results we were able to show that the protein levels of SlETR3 and SlETR4 increased during ripening, probably to control ethylene sensitivity. The other receptors and CTRs showed either stable levels that could sustain, or decreasing levels that could promote fruit ripening.

Ethylene, the plant ripening hormone of climacteric fruit, is perceived by ethylene receptors which is the first step in the complex ethylene signal transduction pathway. Much progress has been made in elucidating the mechanism of this pathway, but there is still a lot to be done in the proteomic quantification of the main proteins involved, particularly during fruit ripening. This work focuses on the mass spectrometry based identification and quantification of the ethylene receptors (ETRs) and the downstream components of the pathway, CTR-like proteins (CTRs) and ETHYLENE INSENSITIVE 2 (EIN2). We used tomato as a model fruit to study changes in protein abundance involved in the ethylene signal transduction during fruit ripening. In order to detect and quantify these low abundant proteins located in the membrane of the endoplasmic reticulum, we developed a workflow comprising sample fractionation and MS analysis using parallel reaction monitoring. This work shows the feasibility of the identification and absolute quantification of all seven ethylene receptors, three out of four CTRs and EIN2 in four ripening stages of tomato. In parallel, gene expression was analyzed through real-time qPCR. Correlation between transcriptomic and proteomic profiles during ripening was only observed for three of the studied proteins, suggesting that the other signaling proteins are likely post-transcriptionally regulated. Based on our quantification results we were able to show that the protein levels of SlETR3 and SlETR4 increased during ripening, probably to control ethylene sensitivity. The other receptors and CTRs showed either stable levels that could sustain, or decreasing levels that could promote fruit ripening. [ABSTRACT FROM AUTHOR]

Durian ( Durio zibethinus ) is a climacteric fruit. Biochemical changes take place in the durian pulp while the husk is dehiscent during fruit ripening. Fruit development of durian cv. Monthong showed a sigmoid curve and reached a maturity stage week 16 after anthesis, while fruit dehiscence started a week after harvest, was hastened by ethephon treatment, and delayed by 1-methylcyclopropene (1-MCP). 1-MCP treatment delayed dehiscence for 5 days from day 7 to day 12 compared with control fruit at 25 °C. Two ethylene receptors ( DzETR1 and DzETR2 ), a CTR1-like protein ( DzCTR1 ) and two ethylene insensitive-like proteins ( DzEIL1 and DzEIL2 ) in the pulp and husk shared high homology to Theobroma cacao . The transcript levels of DzETR2, DzCTR1 and DzEIL2 slightly decreased during fruit development (week 2 to week 10), while DzETR2 transcript level showed the highest expression. The gene expression of DzETR2 in the dehiscence zone, increased at day 12 after harvest, while gene expression of DzETR1, DzCTR1 and DzEIL2 slightly increased after harvest. All genes were reduced in expression with 1-MCP treatment. After that, an increase in expression was found at day 9, followed by a gradual decrease until day 12. These results suggest that DzETR2 plays a role in fruit development and dehiscence of ripening durian. [ABSTRACT FROM AUTHOR]

Drought frequently occurs during wheat (Triticum aestivum L.) grain filling. The objectives of this study were (i) to investigate the effect of post-anthesis drought on programmed cell death (PCD) in wheat endosperm cells and (ii) to examine the role of ethylene (ETH) receptors and abscisic acid (ABA) in regulating wheat endosperm PCD. Two winter wheat cultivars (‘Xindong 18’ and ‘Xindong 22’) were used in this study. Grain samples were collected from normal and drought stressed plants at 5-day intervals between 5 and 35 days post-anthesis. The samples were then compared with respect to cell viability, nuclear morphometry, cell ultrastructure, DNA integrity, nucleic acid content, and nuclease activity. Analysis was also conducted about gene transcripts related to PCD, ETH receptors, and ABA biosynthesis and degradation. Drought stress reduced cell viability, accelerated nuclear deformation, and increased mitochondrial dissolution. The activity of nucleic acid hydrolase was greater, and the nucleic acid concentrations were less in the drought treatments than in the control. As a result, the peak in DNA fragmentation occurred earlier in the drought treatment. Drought stress significantly increased the expression of four genes related to ABA (nced1, nced2, ao1, ao2). In contrast, drought significantly reduced the expression of four genes related to ETH receptors (ers1, ers2 etr1, etr2) and one gene related to PCD (dad1). In summary, the results indicated that drought stress caused PCD to occur earlier in the endosperm of winter wheat. [ABSTRACT FROM AUTHOR]

Ethylene response factor (ERF) has been widely studied in regulating fruit ripening in tomato, apple, banana and kiwifruit, but little is known in pear. In this study 1‐methylcyclopropene (1‐MCP) treatment, an inhibitor of ethylene perception, was conducted at approximately 30 days before harvest to delay fruit ripening in a climacteric white pear cultivar Yali. Transcriptome libraries were constructed and sequenced in pre‐ripening, ripening, and 1‐MCP treated fruits. Data analysis showed that 73 candidate genes related to fruit ripening were induced by 1‐MCP, among which two were positively related, namely 1‐aminocyclopropane‐1‐carboxyla oxidase and an ERF gene (designated as ACO54 and ERF24). Transient transformations in pear fruit revealed that over‐expression of ACO54 enhance transcription level of ERF24 and most ripening‐related genes. Meanwhile, over‐expression of ERF24 raises expression level of ACO54 and partially ripening‐related genes. Moreover, dual‐luciferase and yeast‐one‐hybrid assays unravel an interaction between ERF24 and the ACO54 promoter. Therefore, the ERF24 could directly regulate ACO54 expression by binding to its promoter. These results suggested that the first identified ERF24 is involved in regulating fruit ripening in Chinese white pear. [ABSTRACT FROM AUTHOR]

Rapid fruit ripening is a significant problem that limits the shelf life of durian, with ethylene having a major impact on the regulation of this event. Durian treated with ethephon ripened 3 d after treatment with increased pulp total soluble solids, ethylene production of the whole fruit and decreased pulp firmness compared to the control fruit. 1-MCP treatment delayed ripening by up to 9 d with inhibited accumulation of total soluble solids, color change, softening and ethylene production. Genes related to ethylene perception ( DzETR1 and DzETR2 ) and the signaling pathway ( DzCTR1, DzEIL1 and DzEIL2 ) in the pulp were investigated during this process, using qPCR to quantify changes in gene transcription. All candidate genes were significantly up-regulated in ripening durian pulp. Ethephon treatment increased the expression of DzETR1 and DzETR2 genes, while expression of DzCTR1, DzEIL1 and DzEIL2 were slightly affected. 1-MCP treatment significantly inhibited the expression of the DzETR2 and DzEIL1 genes. The promoters of DzETR2 genes were isolated and their activation by fruit transcription factors studied using transient expression in tobacco leaves. It was found that members of the kiwifruit and apple EIL1, EIL2 and EIL3 genes strongly activated the DzETR2 promoter. These results suggest that ethylene-induced ripening of durian is via the regulation of DzETR2 by EIL transcription factors. [ABSTRACT FROM AUTHOR]

Brazil is a major producer and consumer of various banana types. Thap Maeo is a promising cultivar for the market due to its resistance to Black and Yellow Sigatoka disease. However, a lack of information and postharvest technologies concerning Thap Maeo physiology seems to be a significant problem limiting its expansion in the market. Thus, this study aims to establish Thap Maeo fruit's physical, biochemical, and physiological aspects, defining the best ethylene dosage for treatment considering fruit ripening parameters. Bananas were harvested and monitored during both natural and ethylene-induced ripening processes. Assessments of pulp firmness, peel color and endogenous ethylene production showed different profiles between ethylene-treated and non-treated fruits, whereas the ethylene responses concerning the carbohydrates and hormones profiles, as well as the ethylene receptors expression, were observed in all ethylene-treated fruits, even applying low concentrations of the hormone. It thus indicated the high ethylene-sensitivity of Thap Maeo cultivar. Such postharvest behaviors reverberate in lower ethylene requirements for treatment, which was established at 10 μL L − 1 . Ethylene-inducible changes in fruit volatile compounds throughout ripening are also discussed. [ABSTRACT FROM AUTHOR]

Ethylene signaling is decisive for many plant developmental processes. Among these, control of senescence, abscission and fruit ripening are of fundamental relevance for global agriculture. Consequently, detailed knowledge of the signaling network along with the molecular processes of signal perception and transfer are expected to have high impact on future food production and agriculture. Recent advances in ethylene research have demonstrated that signaling of the plant hormone critically depends on the interaction of the ethylene receptor family with the NRAMP-like membrane protein ETHYLENE INSENSITIVE 2 (EIN2) at the ER membrane, phosphorylation-dependent proteolytic processing of ER-localized EIN2 and subsequent translocation of the cleaved EIN2 C-terminal polypeptide (EIN2-CEND) to the nucleus. EIN2 nuclear transport, but also interaction with the receptors sensing the ethylene signal, both, depend on a nuclear localization signal (NLS) located at the EIN2 C-terminus. Loss of the tight interaction between receptors and EIN2 affects ethylene signaling and impairs plant ethylene responses. Synthetic peptides derived from the NLS sequence interfere with the EIN2–receptor interaction and have utility in controlling plant ethylene responses such as ripening. Here, we report that a synthetic peptide (NOP-1) corresponding to the NLS motif of Arabidopsis EIN2 (aa 1262–1269) efficiently binds to tomato ethylene receptors LeETR4 and NR and delays ripening in the post-harvest phase when applied to the surface of sampled green fruits pre-harvest. In particular, degradation of chlorophylls was delayed by several days, as monitored by optical sensors and confirmed by analytical methods. Similarly, accumulation of β-carotene and lycopene in the fruit pulp after NOP-1 application was delayed, without having impact on the total pigment concentration in the completely ripe fruits. Likewise, the peptide had no negative effects on fruit quality. Our molecular and phenotypic studies reveal that peptide biologicals could contribute to the development of a novel family of ripening inhibitors and innovative ripening control in climacteric fruit.

It has long been known that the gaseous plant hormone ethylene plays a key role in nodulation in legumes. The perception of ethylene by a family of five membrane-localized receptors is necessary to trigger the ethylene signaling pathway, which regulates various biological responses in Arabidopsis. However, a systematic analysis of the ethylene receptors in leguminous plants and their roles in nodule development is lacking. In this study, we performed a characterization of ethylene receptor genes based on the latest Glycine max genome sequence and a public microarray database. Eleven ethylene receptor family genes were identified in soybean through homology searches, and they were divided into two subgroups. Exon–intron analysis showed that the gene structures are highly conserved within each group. Further analysis of their expression patterns showed that these ethylene receptor genes are differentially expressed in various soybean tissues and organs, including functional nodules. Notably, the ethylene receptor genes showed different responses to rhizobial infection and Nod factors, suggesting a possible role for ethylene receptors and ethylene signaling in rhizobia–host cell interactions and nodulation in soybean. Together, these data indicate the functional divergence of ethylene receptor genes in soybean, and that some of these receptors mediate nodulation, including rhizobial infection, nodule development, and nodule functionality. These findings provide a foundation for further elucidation of the molecular mechanism by which the ethylene signaling pathway regulates nodulation in soybean, as well as other legumes.

Ethylene is an essential regulator of fruit development. However, the role of this phytohormone in the development and maturation of peach ( Prunus persica L. Batsch) fruit has not been systematically examined. In this study, through BLAST analysis of tomato ( Solanum lycopersicum ) protein sequences against the peach genome database and transcriptome analysis of all APETALA2/Ethylene Responsive Factor (AP2/ERF) genes in peach, we identified 15 ethylene signal transduction genes and 15 AP2/ERF genes and examined their expression levels in SH (stony hard) peaches and MF (melting flesh) peaches during fruit maturation. We investigated the expression patterns of six genes, including one ETR-like gene (Prupe.1G034300) and five AP2/ERF genes (Prupe.1G037700, Prupe.2G289500, Prupe.3G240000, Prupe.5G061800, and Prupe.7G194400), through quantitative RT-PCR expression analysis. The transcript levels of these genes were lower at the early stages of fruit development and increased dramatically at stage S4 in MF peaches, while they remained low in SH peaches, which mirrors the pattern of changes observed in ethylene production. Treatment of MF (‘Zhongyoutao 13’, ‘CN13’) fruit at stage S4 II with 1-MCP was ineffective in blocking ethylene production, and the expression of these genes was inhibited by 1-MCP treatment in ‘CN13’ on day 1, further demonstrating that these genes are closely related to fruit ripening. We detected a slight change in fruit firmness in ‘CN13’ after 1-MCP treatment, whereas ethylene production was not affected. By contrast, the transcript profile of gene Prupe.5G090800 exhibited the opposite trend. The comprehensive data generated in this study will improve our understanding of the ethylene receptor genes and ERF genes in peach and lay the foundation for further exploring the mechanisms underlying peach fruit ripening and softening. [ABSTRACT FROM AUTHOR]

Ethylene signaling is decisive for many plant developmental processes. Among these, control of senescence, abscission and fruit ripening are of fundamental relevance for global agriculture. Consequently, detailed knowledge of the signaling network along with the molecular processes of signal perception and transfer are expected to have high impact on future food production and agriculture. Recent advances in ethylene research have demonstrated that signaling of the plant hormone critically depends on the interaction of the ethylene receptor family with the NRAMP-like membrane protein ETHYLENE INSENSITIVE 2 (EIN2) at the ER membrane, phosphorylation-dependent proteolytic processing of ER-localized EIN2 and subsequent translocation of the cleaved EIN2 C-terminal polypeptide (EIN2-CEND) to the nucleus. EIN2 nuclear transport, but also interaction with the receptors sensing the ethylene signal, both, depend on a nuclear localization signal (NLS) located at the EIN2 C-terminus. Loss of the tight interaction between receptors and EIN2 affects ethylene signaling and impairs plant ethylene responses. Synthetic peptides derived from the NLS sequence interfere with the EIN2-receptor interaction and have utility in controlling plant ethylene responses such as ripening. Here, we report that a synthetic peptide (NOP-1) corresponding to the NLS motif of Arabidopsis EIN2 (aa 1262-1269) efficiently binds to tomato ethylene receptors LeETR4 and NR and delays ripening in the post-harvest phase when applied to the surface of sampled green fruits pre-harvest. In particular, degradation of chlorophylls was delayed by several days, as monitored by optical sensors and confirmed by analytical methods. Similarly, accumulation of b-carotene and lycopene in the fruit pulp after NOP-1 application was delayed, without having impact on the total pigment concentration in the completely ripe fruits. Likewise, the peptide had no negative effects on fruit quality. Our molecular and phenotypic studies reveal that peptide biologicals could contribute to the development of a novel family of ripening inhibitors and innovative ripening control in climacteric fruit. [ABSTRACT FROM AUTHOR]

The design and construction of a laboratory-scale apparatus for generating variable concentrations and flow rates of exogenous ethylene for fresh produce supplementation during storage trials is described. A stock of compressed ethylene in nitrogen (5000 μl l −1 ) was blended into a continuous flow stream of air and diluted to the desired concentrations. The ethylene and air flow rates were controlled with calibrated mass flow control valves. An empirical mathematical model was derived for real-time variation of both the mixed concentration and flow rate during continuous flow. Validation of the model was performed using fresh sweet potato as a case study where a steady continuous ethylene concentration of 10 μl l −1 was achieved for three months. The bespoke system offers easy-to-manage ethylene supplementation for research. [ABSTRACT FROM AUTHOR]

Copper ions play an important role in ethylene receptor biogenesis and proper function. The copper transporter RESPONSIVE-TO-ANTAGONIST1 (RAN1) is essential for copper ion transport in Arabidopsis thaliana. However it is still unclear how copper ions are delivered to RAN1 and how copper ions affect ethylene receptors. There is not a specific copper chelator which could be used to explore these questions. Here, by chemical genetics, we identified a novel small molecule, triplin, which could cause a triple response phenotype on dark-grown Arabidopsis seedlings through ethylene signaling pathway. ran1-1 and ran1-2 are hypersensitive to triplin. Adding copper ions in growth medium could partially restore the phenotype on plant caused by triplin. Mass spectrometry analysis showed that triplin could bind copper ion. Compared to the known chelators, triplin acts more specifically to copper ion and it suppresses the toxic effects of excess copper ions on plant root growth. We further showed that mutants of ANTIOXIDANT PROTEIN1 (ATX1) are hypersensitive to tiplin, but with less sensitivity comparing with the ones of ran1-1 and ran1-2. Our study provided genetic evidence for the first time that, copper ions necessary for ethylene receptor biogenesis and signaling are transported from ATX1 to RAN1. Considering that triplin could chelate copper ions in Arabidopsis, and copper ions are essential for plant and animal, we believe that, triplin not only could be useful for studying copper ion transport of plants, but also could be useful for copper metabolism study in animal and human. [ABSTRACT FROM AUTHOR]

Multiple environmental stresses affect growth and development of plants. Plants try to adapt under these unfavorable condition through various evolutionary mechanisms like physiological and biochemical alterations connecting various network of regulatory processes. Transcription factors (TFs) like APETALA2/ETHYLENE RESPONSE FACTORS (AP2/ERFs) are an integral component of these signaling cascades because they regulate expression of a wide variety of down stream target genes related to stress response and development through different mechanism. This downstream regulation of transcript does not always positively or beneficially affect the plant but also they display some developmental defects like senescence and reduced growth under normal condition or sensitivity to stress condition. Therefore, tight auto/cross regulation of these TFs at transcriptional, translational and domain level is crucial to understand. The present manuscript discuss the multiple regulation and advantage of plasticity and specificity of these family of TFs to a wide or single downstream target(s) respectively. We have also discussed the concern which comes with the unwanted associated traits, which could only be averted by further study and exploration of these AP2/ERFs. [ABSTRACT FROM AUTHOR]

The conversion of cyclohexanone and substituted cyclohexanones into alkyl aryl ethers by using a Pd/C–ethylene system is discussed, with ethylene functioning as a hydrogen acceptor. The ether products are easily transformed into the corresponding phenols by treatment with BBr3. The direct conversion of cyclohexenone into phenol in the presence of a catalytic amount of Pd/C under an ethylene atmosphere is also described. The conversion of cyclohexanone and substituted cyclohexanones into alkyl aryl ethers by using a Pd/C–ethylene system is disclosed. The ether products are easily transformed into the corresponding phenols by treatment with BBr3. The direct conversion of cyclohexenone into phenol is also described. [ABSTRACT FROM AUTHOR]

Exogenous ethylene, or ethephon, has been widely used to induce pineapple flowering, but the molecular mechanism behind ethephon induction is still unclear. In this study, we cloned four genes encoding ethylene receptors (designated AcERS1a, AcERS1b, AcETR2a and AcETR2b). The 5′ flanking sequences of these four genes were also cloned by self-formed adaptor PCR and SiteFinding-PCR, and a group of putative cis-acting elements was identified. Phylogenetic tree analysis indicated that AcERS1a, AcERS1b, AcETR2a and AcETR2b belonged to the plant ERS1s and ETR2/EIN4-like groups. Quantitative real-time PCR showed that AcETR2a and AcETR2b (subfamily 2) were more sensitive to ethylene treatment compared with AcERS1a and AcERS1b (subfamily 1). The relative expression of AcERS1b, AcETR2a and AcETR2b was significantly increased during the earlier period of pineapple inflorescence formation, especially at 1-9 days after ethylene treatment (DAET), whereas AcERS1a expression changed less than these three genes. In situ hybridization results showed that bract primordia (BP) and flower primordia (FP) appeared at 9 and 21 DAET, respectively, and flowers were formed at 37 DAET. AcERS1a, AcERS1b, AcETR2a and AcETR2b were mainly expressed in the shoot apex at 1-4 DAET; thereafter, with the appearance of BP and FP, higher expression of these genes was found in these new structures. Finally, at 37 DAET, the expression of these genes was mainly focused in the flower but was also low in other structures. These findings indicate that these four ethylene receptor genes, especially AcERS1b, AcETR2a and AcETR2b, play important roles during pineapple flowering induced by exogenous ethephon.

Fruitlet abscission of mango is typically very severe, causing considerable production losses worldwide. Consequently, a detailed physiological and molecular characterization of fruitlet abscission in mango is required to describe the onset and time-dependent course of this process. To identify the underlying key mechanisms of abscission, ethephon, an ethylene releasing substance, was applied at two concentrations (600 ppm, 7200 ppm) during the midseason drop stage of mango. The abscission process is triggered by ethylene diffusing to the abscission zone where it binds to specific receptors and thereby activating several key physiological responses at the cellular level. The treatments reduced significantly the capacity of polar auxin transport through the pedicel at one day after treatment and thereafter when compared to untreated pedicels. The transcript levels of the ethylene receptor genes MiETR1 and MiERS1 were significantly upregulated in the pedicel and pericarp at one, two and three days after the ethephon application with 7200 ppm, except for MiETR1 in the pedicel, when compared to untreated fruitlet. In contrast, ethephon applications with 600 ppm did not affect expression levels of MiETR1 in the pedicel and of MiERS1 in the pericarp; however, MiETR1 in the pericarp at day two and MiERS1 in the pedicel at days two and three were significantly upregulated over the controls. Moreover, two novel short versions of the MiERS1 were identified and detected more often in the pedicel of treated than untreated fruitlets at all sampling times. Sucrose concentration in the fruitlet pericarp was significantly reduced to the control at two days after both ethephon treatments. In conclusion, it is postulated that the ethephon-induced abscission process commences with a reduction of the polar auxin transport capacity in the pedicel, followed by an upregulation of ethylene receptors and finally a decrease of the sucrose concentration in the fruitlets.

Tomatoes with a prolonged fruit shelf life and improved postharvest quality would be an attractive commodity for both breeders and consumers. A weak allele of the tomato ethylene receptor mutant Sletr1-2 elicits an extended fruit shelf life without prominent undesirable pleiotropic effects. In this study, we elucidate the influences of the Sletr1-2 mutation on alterations of the postharvest fruit quality of Sletr1-2 F1 hybrid lines from four different pure-line cultivar parents. The changes in the compositions of metabolic compounds, including sugars, organic acids and amino acids, over 30 days of postharvest storage have been evaluated. The Sletr1-2 mutation significantly affected the postharvest fruit quality parameters of the Sletr1-2 F1 hybrid lines in a manner that depended on the pure-line cultivar parental backgrounds. The influence of the Sletr1-2 mutation was detected only in the reductions and/or increases of individual amino acids and increases in the levels of organic acids, i.e., malate and citrate. In contrast, the sugar level was not changed. Moreover, the Sletr1-2 mutation significantly reduced the rate of water loss during postharvest storage. These results indicate that the Sletr1-2 mutation has favorable effects on the postharvest changes of Sletr1-2 F1 hybrid tomatoes that improve both the shelf life and the taste quality. [ABSTRACT FROM AUTHOR]

The gaseous plant hormone ethylene, recognized by plant ethylene receptors, plays a pivotal role in various aspects of plant growth and development. ETHYLENE RESPONSE1 (ETR1) is an ethylene receptor isolated from Arabidopsis and has a structure characteristic of prokaryotic two-component histidine kinase (HK) and receiver domain (RD), where the RD structurally resembles bacteria response regulators (RRs). The ETR1 HK domain has autophosphorylation activity, and little is known if the HK can transfer the phosphoryl group to the RD for receptor signaling. Unveiling the correlation of the receptor structure and phosphorylation status would advance the studies towards the underlying mechanisms of ETR1 receptor signaling. In this study, using the nuclear magnetic resonance technique, our data suggested that the ETR1-RD is monomeric in solution and the rigid structure of the RD prevents the conserved aspartate residue phosphorylation. Comparing the backbone dynamics with other RRs, we propose that backbone flexibility is critical to the RR phosphorylation. Besides the limited flexibility, ETR1-RD has a unique γ loop conformation of opposite orientation, which makes ETR1-RD unfavorable for phosphorylation. These two features explain why ETR1-RD cannot be phosphorylated and is classified as an atypical type RR. As a control, phosphorylation of the ETR1-RD was also impaired when the sequence was swapped to the fragment of the bacterial typical type RR, CheY. Here, we suggest a molecule insight that the ETR1-RD already exists as an active formation and executes its function through binding with the downstream factors without phosphorylation. [ABSTRACT FROM AUTHOR]

The aim of this study is to investigate the inhibitory effect of 1-octylcyclopropene (1-OCP), a structural analog of 1-methylcyclopropene (1-MCP), on postharvest ripening and senescence of tomato fruit ( Solanum lycopersicum Mill., Shenyang, China). The results show that 1-OCP is effective in postponing the occurrence time of ethylene production peak and respiration rate peak, delaying the softening and color change, inhibiting the increasing of soluble solids and the decreasing of titratable acidity, and suppressing the expression of LeETR 1 and LeETR 4 in tomatoes. In addition, 1.2 μL L −1 1-OCP is found to be most effective in delaying ripening and senescence process of tomatoes by inhibiting the activities of enzymes involved in ethylene biosynthesis and gene expression of ethylene receptors. It is suggested that 1-OCP is potential as a given inhibitor when applied before the onset of ethylene action which is crucial when considering the promising candidates for practical use. [ABSTRACT FROM AUTHOR]

Physiological changes associated with senescence of flowers and abscission of floral parts in Oncidesa (formerly Oncidium) cv. Gower Ramsey are caused by a plant hormone ethylene which is produced by pollinia cap dislodgment during postharvest handling and transportation. The ethylene receptor gene OgERS1 of Oncidesa has been previously cloned and characterized. To analyze promoter activity of OgERS1, transgenic Arabidopsis thaliana plants were generated to express the ß-glucuronidase (GUS) reporter gene under the control of 5'-upstream sequence of OgERS1 from Oncidesa. The expression pattern of the OgERS1 promoter at the cellular level was investigated by analysis of GUS activity. This promoter can activate gene expression in both actively dividing young tissues and abscission-related aging tissues. Expression of GUS was detected in the shoot meristem uniquely in 10 to 30 d-old-plants and was found in flower buds, axillary buds, flower stems, and abscission layers during later development. In 2- to 3-week-old transgenic Arabidopsis, exogenous ethylene, glucose, lactose, and maltose enhanced promoter activity implying that crosstalk between sugar and an ethylene receptor may exist. However, indole-3-acetic acid, benzylaminopurine, abscisic acid, heat, wounding, salinity, drought, and flooding slightly suppressed promoter activity. These results demonstrate that the promoter of OgERS1 was developmentally and environmentally regulated, and imply a potential for application of this bi-functional promoter to increase branching or enhanced dwarfing. [ABSTRACT FROM AUTHOR]

Different versions of the mango ethylene receptor MiERS1 were identified and the analysis indicates that, in addition to MiERS1 , two short versions of this receptor ( MiERS1m , MiERS1s ), representing truncated proteins with central deletions of functional domains, are present in mango. The short receptor versions reveal a different expression pattern compared to MiERS1 , and they are highly variably transcribed. With transient expression assays using fluorescent fusion proteins, the localisation and the interaction of the receptors were determined in leaf cells of the tobacco model. MiERS1, MiETR1, and the short MiERS1 receptor versions are anchored in the endoplasmic reticulum (ER) membrane and co-localise with each other and with an ER-marker. Furthermore, ectopic expression of the mango receptors appears to induce a re-organisation of the ER resulting in accumulation of ER bodies. Interaction assays suggest that both short MiERS1 receptor versions can bind to proteins located in the ER. Bi-molecular fluorescence complementation (BiFC) assays indicate, that MiERS1m may dimerise with itself and can also interact with MiERS1, but not with MiETR1. Further, it as found that MiETR1 can interact with MiERS1. Interaction of MiERS1s with the other ethylene receptors could not be detected, although it was located in the ER membrane system. [ABSTRACT FROM AUTHOR]

An ethylene receptor gene named BoERS1 was cloned from a bamboo ( Bambusa oldhamii) cDNA library. The open reading frame of BoERS1 was 1,899 bp and encoded a 632-amino acid protein, which contains the five conserved motifs (H, N, G1, F, and G2 boxes) of the bacterial two-component system histidine kinases and shows high sequence similarity with other ethylene receptors in plants, such as rice and maize. Expression of BoERS1 in bamboo shoots increased with the growth of the emerging shoots. In an in vitro kinase assay, the expressed histidine kinase domain of BoERS1 (BHK) was phosphorylated in the presence of Mn, and LC-ESI-MS/MS analysis showed that four amino acids, namely T442, S444, S489, and S503, were phosphorylated. It is interesting to note that S489 and S503 are located in a loop region (L1) that is found only in plant histidine kinase-containing enzymes. The identification of multiple phosphorylation sites on BoERS1 provides a new avenue for future structure-function studies of the ethylene receptor protein family. [ABSTRACT FROM AUTHOR]

The immature non-climacteric fruit of zucchini is very sensitive to postharvest chilling injury (PCI). Although the fruit produces very low ethylene at harvest, cold storage induces an increase in ethylene production in the refrigerated fruit, and upon rewarming up to room temperature. The production of ethylene after rewarming was proportional to cold damage and found to be higher in the fruit of the cultivars most susceptible to chilling. The effects of the ethylene inhibitor 1-methycyclopropene (1-MCP) has been analyzed using the postharvest quality parameters of nine zucchini cultivars stored at 20 °C and 4 °C for 14 days. We have also determined the evolution of ethylene production in the fruit of these cultivars, and assessed the expression profiles of twelve ethylene biosynthesis and signaling genes in both control fruit and in 1-MCP treated fruit, of two cultivars that responded to 1-MCP. The 1-MCP treatment reduced the rate and delayed the onset of PCI symptoms and reduced fruit weight loss, although only in the fruit of the most chilling-susceptible cultivars, and concomitantly we found a reduction in the respiration rate and in the level of cold-induced ethylene. This data suggests that ethylene is not only a response of the fruit to cold damage, as occurs upon rewarming, but could also play a regulation role in the onset of PCI in this non-climacteric fruit. The genes CpACO1 and CpACS1 were found to be involved in the biosynthesis of the cold-induced ethylene, but the role played by CpACO1 was found to be the key function, since it was induced higher than CpACS1 in response to cold storage. More over CpACO1 was the only ethylene biosynthesis gene that was downregulated by the 1-MCP treatment. Some of the ethylene perception and signaling genes, including CpETR1 , CpCTR1 and CpEIN3.1 were also induced in response to cold storage, and were downregulated in response to 1-MCP, indicating that the positive effect of 1-MCP in the fruit of certain zucchini cultivars is also associated with a downregulation of genes involved in ethylene perception and signal transduction pathways. [ABSTRACT FROM AUTHOR]

Despite the prevalence of poor grain filling in rice (Oryza sativa L.) under abiotic stress, the reason for this is largely unexplored. Application of 0.75% NaCl to a salt-sensitive rice cultivar at late booting resulted in a >20% yield loss. Spikelets per panicle and the percentage of filled grain decreased significantly in response to NaCl application. The inhibitory effect of NaCl on grain filling was greater in basal than in apical spikelets. Sucrose synthase (SUS) activity was positively correlated with grain weight. The transcript levels of the SUS isoforms differed greatly: the levels of SUS2 increased significantly in response to salt; those of SUS4 decreased drastically. Gene expression studies of starch synthase and ADP-glucose pyrophosphorylase showed that the decreased transcript levels of one isoform was compensated by an increase in those of the other. Salt application also significantly increased the gene expression of the ethylene receptors and the ethylene signalling proteins. The increase in their transcript levels was comparatively greater in basal than in apical spikelets. Significant enhancement in the transcript levels of the ethylene receptors and the increase in the production of ethylene indicated that the salt-induced inhibition of grain filling might be mediated by ethylene. Additionally, the inhibition of chromosomal endoreduplication mediated by decreased transcript levels of B-type cyclin could explain poor grain filling under salt stress. A significant increase in the transcript levels of the ethylene-responsive factors in the spikelets during grain filling in response to salt indicated their possible protective role in grain filling under stress. [ABSTRACT FROM AUTHOR]

The salt stress triggered by sublethal, 100 mM and lethal, 250 mM NaCl induced ethylene production as well as rapid accumulation of superoxide radical and H 2 O 2 in the root tips of tomato ( Solanum lycopersicum cv. Ailsa Craig) wild type and ethylene receptor mutant, Never ripe ( Nr/Nr) plants. In the wild type plants superoxide accumulation confined to lethal salt concentration while H 2 O 2 accumulated more efficiently under sublethal salt stress. However, in Nr roots the superoxide production was higher and unexpectedly, H 2 O 2 level was lower than in the wild type under sublethal salt stress. Nitric oxide production increased significantly under sublethal and lethal salt stress in both genotypes especially in mutant plants, while peroxynitrite accumulated significantly under lethal salt stress. Thus, the nitro-oxidative stress may be stronger in Nr roots, which leads to the programmed death of tissues, characterized by the DNA and protein degradation and loss of cell viability under moderate salt stress. In Nr mutants the cell death was induced in the absence of ethylene perception. Although wild type roots could maintain their potassium content under moderate salt stress, K + level significantly declined leading to small K + /Na + ratio in Nr roots. Thus Nr mutants were more sensitive to salt stress than the wild type and the viability of root cells decreased significantly under moderate salt stress. These changes can be attributed to a stronger ionic stress due to the K + loss from the root tissues. [ABSTRACT FROM AUTHOR]

The lace plant, Aponogeton madagascariensis, is an aquatic monocot that forms perforations in its leaves as part of normal leaf development. Perforation formation occurs through developmentally regulated programmed cell death (PCD). The molecular basis of PCD regulation in the lace plant is unknown, however ethylene has been shown to play a significant role. In this study, we examined the role of ethylene receptors during perforation formation. We isolated three lace plant ethylene receptors AmERS1a, AmERS1b and AmERS1c. Using quantitative PCR, we examined their transcript levels at seven stages of leaf development. Through laser-capture microscopy, transcript levels were also determined in cells undergoing PCD and cells not undergoing PCD (NPCD cells). AmERS1a transcript levels were significantly lower in window stage leaves (in which perforation formation and PCD are occurring) as compared to all other leaf developmental stages. AmERS1a and AmERS1c (the most abundant among the three receptors) had the highest transcript levels in mature stage leaves, where PCD is not occurring. Their transcript levels decreased significantly during senescence-associated PCD. AmERS1c had significantly higher transcript levels in NPCD compared to PCD cells. Despite being significantly low in window stage leaves, AmERS1a transcripts were not differentially expressed between PCD and NPCD cells. The results suggested that ethylene receptors negatively regulate ethylene-controlled PCD in the lace plant. A combination of ethylene and receptor levels determines cell fate during perforation formation and leaf senescence. A new model for ethylene emission and receptor expression during lace plant perforation formation and senescence is proposed. [ABSTRACT FROM AUTHOR]

Apple (Malus × domestica) is increasingly being considered an interesting model species for studying early fruit development, during which an extremely relevant phenomenon, fruitlet abscission, may occur as a response to both endogenous and/or exogenous cues. Several studies were carried out shedding light on the main physiological and molecular events leading to the selective release of lateral fruitlets within a corymb, either occurring naturally or as a result of a thinning treatment. Several studies pointed out a clear association between a rise of ethylene biosynthetic levels in the fruitlet and its tendency to abscise. A direct mechanistic link, however, has not yet been established between this gaseous hormone and the generation of the abscission signal within the fruit. In this work, the role of ethylene during the very early stages of abscission induction was investigated in fruitlet populations with different abscission potentials due either to the natural correlative inhibitions determining the so-called physiological fruit drop or to a well-tested thinning treatment performed with the cytokinin benzyladenine. A crucial role was ascribed to the ratio between the ethylene produced by the cortex and the expression of ethylene receptor genes in the seed. This ratio would determine the final probability to abscise. A working model has been proposed consistent with the differential distribution of four receptor transcripts within the seed, which resembles a spatially progressive cell-specific immune-like mechanism evolved by apple to protect the embryo from harmful ethylene. [ABSTRACT FROM AUTHOR]

The legume-rhizobium symbiosis is initiated through the activation of the Nodulation (Nod) factor-signaling cascade, leading to a rapid reprogramming of host cell developmental pathways. In this work, we combine transcriptome sequencing with molecular genetics and network analysis to quantify and categorize the transcriptional changes occurring in roots of Medicago truncatula from minutes to days after inoculation with Sinorhizobium medicae. To identify the nature of the inductive and regulatory cues, we employed mutants with absent or decreased Nod factor sensitivities (i.e. Nodulation factor perception and Lysine motif domain-containing receptor-like kinase3, respectively) and an ethylene (ET)-insensitive, Nod factor-hypersensitive mutant (sickle). This unique data set encompasses nine time points, allowing observation of the symbiotic regulation of diverse biological processes with high temporal resolution. Among the many outputs of the study is the early Nod factorinduced, ET-regulated expression of ET signaling and biosynthesis genes. Coupled with the observation of massive transcriptional derepression in the ET-insensitive background, these results suggest that Nod factor signaling activates ET production to attenuate its own signal. Promoter:b-glucuronidase fusions report ET biosynthesis both in root hairs responding to rhizobium as well as in meristematic tissue during nodule organogenesis and growth, indicating that ET signaling functions at multiple developmental stages during symbiosis. In addition, we identified thousands of novel candidate genes undergoing Nod factor-dependent, ET-regulated expression. We leveraged the power of this large data set to model Nod factor- and ET-regulated signaling networks using MERLIN, a regulatory network inference algorithm. These analyses predict key nodes regulating the biological process impacted by Nod factor perception. We have made these results available to the research community through a searchable online resource. [ABSTRACT FROM AUTHOR]

Ethylene influences the growth and development of Arabidopsis (Arabidopsis thaliana) via five receptor isoforms. However, the ETHYLENE RESPONSE1 (ETR1) ethylene receptor has unique, and sometimes contrasting, roles from the other receptor isoforms. Prior research indicates that the receiver domain of ETR1 is important for some of these noncanonical roles. We determined that the ETR1 receiver domain is not needed for ETR1's predominant role in mediating responses to the ethylene antagonist, silver. To understand the structure-function relationship underlying the unique roles of the ETR1 receiver domain in the control of specific traits, we performed alanine-scanning mutagenesis. We chose amino acids that are poorly conserved and are in regions predicted to have altered tertiary structure compared with the receiver domains of the other two receptors that contain a receiver domain, ETR2 and ETHYLENE INSENSITIVE4. The effects of these mutants on various phenotypes were examined in transgenic, receptor-deficient Arabidopsis plants. Some traits, such as growth in air and growth recovery after the removal of ethylene, were unaffected by these mutations. By contrast, three mutations on one surface of the receiver domain rendered the transgene unable to rescue ethylene-stimulated nutations. Additionally, several mutations on another surface altered germination on salt. Some of these mutations conferred hyperfunctionality to ETR1 in the context of seed germination on salt, but not for other traits, that correlated with increased responsiveness to abscisic acid. Thus, the ETR1 receiver domain has multiple functions where different surfaces are involved in the control of different traits. Models are discussed for these observations. [ABSTRACT FROM AUTHOR]

Fruitlet abscission of mango is typically very severe, causing considerable production losses worldwide. Consequently, a detailed physiological and molecular characterization of fruitlet abscission in mango is required to describe the onset and time-dependent course of this process. To identify the underlying key mechanisms of abscission, ethephon, an ethylene releasing substance, was applied at two concentrations (600 and 7200 ppm) during the midseason drop stage of mango. The abscission process is triggered by ethylene diffusing to the abscission zone where it binds to specific receptors and thereby activating several key physiological responses at the cellular level. The treatments reduced significantly the capacity of polar auxin transport through the pedicel at 1 day after treatment and thereafter when compared to untreated pedicels. The transcript levels of the ethylene receptor genes MiETR1 and MiERS1 were significantly upregulated in the pedicel and pericarp at 1, 2, and 3 days after the ethephon application with 7200 ppm, except for MiETR1 in the pedicel, when compared to untreated fruitlet. In contrast, ethephon applications with 600 ppm did not affect expression levels of MiETR1 in the pedicel and of MiERS1 in the pericarp; however, MiETR1 in the pericarp at day 2 and MiERS1 in the pedicel at days 2 and 3 were significantly upregulated over the controls. Moreover, two novel short versions of the MiERS1 were identified and detected more often in the pedicel of treated than untreated fruitlets at all sampling times. Sucrose concentration in the fruitlet pericarp was significantly reduced to the control at 2 days after both ethephon treatments. In conclusion, it is postulated that the ethephon-induced abscission process commences with a reduction of the polar auxin transport capacity in the pedicel, followed by an upregulation of ethylene receptors and finally a decrease of the sucrose concentration in the fruitlets. [ABSTRACT FROM AUTHOR]

The ethylene is a gaseous hormone which plays multiple roles in regulating plant growth and development and also serves as a key modulator of the plant's response to biotic or abiotic stresses. Its production is tightly regulated by internal signals during development and in response to environmental stimuli from biotic and abiotic stresses. Arabidopsis is a model plant to understand the ethylene signal transduction. In Arabidopsis, ethylene is perceived by a family of five receptors such as ETR1, ETR2, ERS1, ERS2 and EIN4 which are predominantly localized to the ER membrane. Ethylene receptor can be divided into two subfamilies i.e. type I and type II subfamilies, depending on the basis of structural similarities. CTR1 functions as a key mediator of ethylene signal transduction. Ethylene was originally regarded as a stress hormone because its synthesis is induced by a variety of environmental stress signals. Among the environmental stresses, such as ozone, wounding, and UV irradiation are the stimulations for ethylene synthesis. Enhanced ethylene production is an early, active response of plants to perception of pathogen attack and is associated with the induction of defence reactions. The elucidation of the mechanisms of ethylene signal transduction in response to biotic and abiotic stress is providing a framework for understanding how all plants sense and respond to ethylene. [ABSTRACT FROM AUTHOR]

Background: Ethylene plays critical roles in plant growth and development, including the regulation of cell expansion, senescence, and the response to biotic and abiotic stresses. Elements of the initial signal transduction pathway have been determined, but we are still defining regulatory mechanisms by which the sensitivity of plants to ethylene is modulated. Results: We report here that members of the ARGOS gene family of Arabidopsis, previously implicated in the regulation of plant growth and biomass, function as negative feedback regulators of ethylene signaling. Expression of all four members of the ARGOS family is induced by ethylene, but this induction is blocked in ethylene-insensitive mutants. The dose dependence for ethylene induction varies among the ARGOS family members, suggesting that they could modulate responses across a range of ethylene concentrations. GFP-fusions of ARGOS and ARL localize to the endoplasmic reticulum, the same subcellular location as the ethylene receptors and other initial components of the ethylene signaling pathway. Seedlings with increased expression of ARGOS family members exhibit reduced ethylene sensitivity based on physiological and molecular responses. Conclusions: These results support a model in which the ARGOS gene family functions as part of a negative feedback circuit to desensitize the plant to ethylene, thereby expanding the range of ethylene concentrations to which the plant can respond. These results also indicate that the effects of the ARGOS gene family on plant growth and biomass are mediated through effects on ethylene signal transduction. [ABSTRACT FROM AUTHOR]

Grain yields in modern super rice cultivars do not always meet the expectations because many spikelets are located on secondary branches in closely packed homogeneous distribution in these plants, and they do not fill properly. The factors limiting grain filling of such spikelets, especially in the lower panicle branches, are elusive. Two long-duration rice cultivars differing in panicle density, Mahalaxmi (compact) and Upahar (lax), were cultivated in an open field plot. Grain filling, ethylene production and constitutive expression of ethylene receptors and ethylene signal transducers in apical and basal spikelets of the panicle were compared during the early post-anthesis stage, which is the most critical period for grain development. In another experiment, a similar assessment was made for the medium-duration cultivars compact-panicle OR-1918 and lax-panicle Lalat. Grain weight of the apical spikelets was always higher than that of the basal spikelets. This gradient of grain weight was wide in the compact-panicle cultivars and narrow in the lax-panicle cultivars. Compared to apical spikelets, the basal spikelets produced more ethylene at anthesis and retained the capacity for post-anthesis expression of ethylene receptors and ethylene signal transducers longer. High ethylene production enhanced the expression of the RSR1 gene, but reduced expression of the GBSS1 gene. Ethylene inhibited the partitioning of assimilates of developing grains resulting in low starch biosynthesis and high accumulation of soluble carbohydrates. It is concluded that an increase in grain/spikelet density in rice panicles reduces apical dominance to the detriment of grain filling by production of ethylene and/or enhanced perception of the ethylene signal. Ethylene could be a second messenger for apical dominance in grain filling. The manipulation of the ethylene signal would possibly improve rice grain yield. [ABSTRACT FROM AUTHOR]

Ethylene receptors are involved in ethylene sensing and signal transduction. This study was conducted to isolate genes encoding ethylene receptors from melon fruit and to determine their expression during postharvest fruit ripening. The amplification of melon fruit cDNA with degenerate primers revealed two partial cDNAs. Upon 3' and 5' RACE analysis of the cDNAs, two full length genes designated Cm-ETR1 and Cm-ETR2 were obtained. Cloning and sequencing of the genes showed that Cm-ETR1 and Cm-ETR2 genes were 2320 and 2537 bp in length, respectively. Cm-ETR1 had a predicted open reading frame (ORF) of 2223 nucleotides encoding a protein of 740 amino acids. Cm-ETR2 contained a predicted ORF of 2301 nucleotides encoding a protein of 766 amino acids. Structure and sequence analyses demonstrated that both genes had conserved domains found in ethylene receptor genes from other species, and were highly similar in terms of both nucleotide and amino acid sequences to the ethylene receptor sequences isolated from other species. The expression analyses of the genes revealed that the transcript levels of both receptor genes significantly increased during postharvest storage. The isolated receptor genes could be used to manipulate ethylene effects to delay the development of ethylene related physiological disorders and enhance the shelf life of melon fruit. [ABSTRACT FROM AUTHOR]

Heterotrimeric G proteins function as key players in hydrogen peroxide (H2O2) production in plant cells, but whether G proteins mediate ethylene-induced H2O2 production and stomatal closure are not clear. Here, evidences are provided to show the Gα subunit GPA1 as a missing link between ethylene and H2O2 in guard cell ethylene signalling. In wild-type leaves, ethylene-triggered H2O2 synthesis and stomatal closure were dependent on activation of Gα. GPA1 mutants showed the defect of ethylene-induced H2O2 production and stomatal closure, whereas wGα and cGα overexpression lines showed faster stomatal closure and H2O2 production in response to ethylene. Ethylene-triggered H2O2 generation and stomatal closure were impaired in RAN1, ETR1, ERS1 and EIN4 mutants but not impaired in ETR2 and ERS2 mutants. Gα activator and H2O2 rescued the defect of RAN1 and EIN4 mutants or etr1-3 in ethylene-induced H2O2 production and stomatal closure, but only rescued the defect of ERS1 mutants or etr1-1 and etr1-9 in ethylene-induced H2O2 production. Stomata of CTR1 mutants showed constitutive H2O2 production and stomatal closure, but which could be abolished by Gα inhibitor. Stomata of EIN2, EIN3 and ARR2 mutants did not close in responses to ethylene, Gα activator or H2O2, but do generate H2O2 following challenge of ethylene or Gα activator. The data indicate that Gα mediates ethylene-induced stomatal closure via H2O2 production, and acts downstream of RAN1, ETR1, ERS1, EIN4 and CTR1 and upstream of EIN2, EIN3 and ARR2. The data also show that ETR1 and ERS1 mediate both ethylene and H2O2 signalling in guard cells. [ABSTRACT FROM AUTHOR]

We studied the expression of a gene encoding an ethylene receptor, called Ethylene Response Sensor 1 ( Den-ERS1 ), in the petals of Dendrobium orchid flowers. Transcripts accumulated during the young floral bud stage and declined by the time the flowers had been open for several days. Pollination or exposure to exogenous ethylene resulted in earlier flower senescence, an increase in ethylene production and a lower Den-ERS1 transcript abundance. Treatment with 1-methylcyclopropene (1-MCP), an inhibitor of the ethylene receptor, decreased ethylene production and resulted in high transcript abundance. The literature indicates two kinds of ethylene receptor genes with regard to the effects of ethylene. One group shows ethylene-induced down-regulated transcription, while the other has ethylene-induced up-regulation. The present gene is an example of the first group. The 5′ flanking region showed binding sites for Myb and myb-like, homeodomain, MADS domain, NAC, TCP, bHLH and EIN3-like transcription factors. The binding site for the EIN3-like factor might explain the ethylene effect on transcription. A few other transcription factors (RAV1 and NAC) seem also related to ethylene effects. [ABSTRACT FROM AUTHOR]

Ethylene receptor (ethylene response sensor, ERS) is the primary component involving in the ethylene biosynthesis and ethylene signal transduction pathway. In the present study, a GZ- ERS gene encoding ERS was cloned from a sugarcane cv. YL17 ( Saccharum spp.) using RT-PCR and ligation-mediated PCR with primers designed based on conserved amino acid sequences of ERS from rice and corn. The GZ- ERS gene was 2,243 base pairs in length and coded for a polypeptide of 633 amino acid residues. The amino acid sequence of GZ- ERS encoded protein was 97, 96 and 91 % identical to OS- ERS ( AAX95525) of rice, ZmERS25 ( AAR25567) of corn, and Sc- ERS ( ACF60981) of a sugarcane cv. ROC22, respectively. Genomic Southern analysis using a GZ- ERS cDNA insert as the probe indicated the presence of a single copy of GZ- ERS gene per haploid sugarcane genome. Real-time qPCR analysis showed that GZ- ERS was widely expressed in the vegetative tissues of sugarcane, and its expression in maturing internodes and leaf tissues was higher than in immature tissues. Furthermore, the expression level of GZ- ERS was the highest in maturing leaves, but very low in immature internodes. The GZ- ERS expression level in maturing leaves was enhanced by ethephon treatment, but was reduced under low temperature, drought, and dark conditions. The results from this study may help understand the regulation of gene expression in sugarcane and its responses to plant growth regulator ethephon and environmental stresses. As the sugarcane ERS family has multiple members, other members need to be isolated and characterized in future studies. [ABSTRACT FROM AUTHOR]