Ethylene Biosynthesis In Plants Research Articles

Functional mechanism study of the allelochemical myrigalone A identifies a group of ultrapotent inhibitors of ethylene biosynthesis in plants

Allelochemicals represent a class of natural products released by plants as root, leaf, and fruit exudates that interfere with the growth and survival of neighboring plants. Understanding how allelochemicals function to regulate plant responses may provide valuable new approaches to better control plant function. One such allelochemical, Myrigalone A (MyA) produced by Myrica gale, inhibits seed germination and seedling growth through an unknown mechanism. Here, we investigate MyA using the tractable model Dictyostelium discoideum and reveal that its activity depends on the conserved homolog of the plant ethylene synthesis protein 1-aminocyclopropane-1-carboxylic acid oxidase (ACO). Furthermore, in silico modeling predicts the direct binding of MyA to ACO within the catalytic pocket. In D. discoideum, ablation of ACO mimics the MyA-dependent developmental delay, which is partially restored by exogenous ethylene, and MyA reduces ethylene production. In Arabidopsis thaliana, MyA treatment delays seed germination, and this effect is rescued by exogenous ethylene. It also mimics the effect of established ACO inhibitors on root and hypocotyl extension, blocks ethylene-dependent root hair production, and reduces ethylene production. Finally, in silico binding analyses identify a range of highly potent ethylene inhibitors that block ethylene-dependent response and reduce ethylene production in Arabidopsis. Thus, we demonstrate a molecular mechanism by which the allelochemical MyA reduces ethylene biosynthesis and identify a range of ultrapotent inhibitors of ethylene-regulated responses.

GhXB38D represses cotton fibre elongation through ubiquitination of ethylene biosynthesis enzymes GhACS4 and GhACO1.

Ethylene plays an essential role in the development of cotton fibres. Ethylene biosynthesis in plants is elaborately regulated by the activities of key enzymes, 1-aminocyclopropane-1-carboxylate oxidase (ACO) and 1-aminocyclopropane-1-carboxylate synthase (ACS); however, the potential mechanism of post-translational modification of ACO and ACS to control ethylene synthesis in cotton fibres remains unclear. Here, we identify an E3 ubiquitin ligase, GhXB38D, that regulates ethylene biosynthesis during fibre elongation in cotton. GhXB38D gene is highly expressed in cotton fibres during the rapid elongation stage. Suppressing GhXB38D expression in cotton significantly enhanced fibre elongation and length, accompanied by the up-regulation of genes associated with ethylene signalling and fibre elongation. We demonstrated that GhXB38D interacts with the ethylene biosynthesis enzymes GhACS4 and GhACO1 in elongating fibres and specifically mediates their ubiquitination and degradation. The inhibition of GhXB38D gene expression increased the stability of GhACS4 and GhACO1 proteins in cotton fibres and ovules, resulting in an elevated concentration of ethylene. Our findings highlight the role of GhXB38D as a regulator of ethylene synthesis by ubiquitinating ACS4 and ACO1 proteins and modulating their stability. GhXB38D acts as a negative regulator of fibre elongation and serves as a potential target for enhancing cotton fibre yield and quality through gene editing strategy.

Transcriptome profiling reveals ethylene formation in rice seeds by trichloroisocyanuric acid.

Ethylene formation via methionine reacting with trichloroisocyanuric acid under FeSO4 condition in a non-enzymatical manner provides one economically and efficiently novel ethylene-forming approach in planta. Rice seed germination can be stimulated by trichloroisocyanuric acid (TCICA). However, the molecular basis of TCICA in stimulating rice seed germination remains unclear. In this study, the molecular mechanism on how TCICA stimulated rice seed germination was examined via comparative transcriptome. Results showed that clustering of transcripts of TCICA-treated seeds, water-treated seeds, and dry seeds was clearly separated. Twenty-two and three hundred differentially expressed genes were identified as TCICA treatment responsive genes and TCICA treatment potentially responsive genes, respectively. Two and one TCICA treatment responsive genes were involved in ethylene signal transduction and iron homeostasis, respectively. Seventeen of the three hundred TCICA treatment potentially responsive genes were significantly annotated to iron ion binding. Meanwhile, level of methionine (ethylene precursor) showed a 73.9% decrease in response to TCICA treatment. Ethylene was then proved to produce via methionine reacting with TCICA under FeSO4 condition in vitro. Revealing ethylene formation by TCICA not only may bring novel insights into crosstalk between ethylene and other phytohormones during rice seed germination, but also may provide one economically and efficiently novel approach to producing ethylene inplanta independently of the ethylene biosynthesis in plants and thereby may broaden its applications in investigational and applied purposes.

Soilborne bacterium Klebsiella pneumoniae promotes cluster root formation in white lupin through ethylene mediation.

Cluster roots of white lupin are induced by low phosphorus (LP) to efficiently access unavailable P, but how soilborne microbes are associated with cluster root formation (CRF) is unclear. We investigated the roles of soilborne bacteria in CRF response to LP by high-throughput sequencing and root-bacteria interactions. Cluster root number was significantly decreased in plants grown in sterilized soil compared with nonsterilized soil. Proteobacteria was enriched in CR, as shown by microbiome analysis of soil (bulk, rhizosphere, and rhizosheath) and roots (main, lateral, and CR). Large-scale gene expression level implicated ethylene mediation in CRF. Klebsiella pneumoniae (P7), a soilborne bacterium belonging to Proteobacteria, was isolated from CR. Among 11 isolated strains, P7 exhibited the highest 1-aminocyclopropane-1-carboxylate deaminase (ACCD) activity; this enzyme inhibits the biosynthesis of ethylene in plants by the cleavage of the ethylene precursor 1-aminocyclopropane-1-carboxylic acid and promotes CRF under LP. We constructed an ACCD-deficit mutant accd in the P7 genetic background. The loss-of-function mutation failed to promote CRF under LP conditions. Also, auxin responses may be involved in K. pneumoniae-ethylene-mediated CRF. Overall, we propose that the soilborne bacterium K. pneumoniae promotes CRF of white lupin in response to LP by ethylene mediation.

Role of salicylic acid in regulating ethylene and physiological characteristics for alleviating salinity stress on germination, growth and yield of sweet pepper.

BackgroundDuring a preliminary study, effects of 0, 20, 40, and 60 mM NaCl salinity were assessed on germination rate in relation to electrolyte leakage (EL) in sweet pepper. Results explored significant rises in ethylene evolution from seeds having more EL. It was, therefore, hypothesized that excessive ethylene biosynthesis in plants due to salinity stress might be a root cause of low crop productivity. As salicylic acid is one of the potent ethylene inhibitors, thus SA was used to combat effects of ethylene produced under salinity stress of 60 mM NaCl on different physiological and morphological characteristics of sweet pepper.MethodologyThe effect of 0.05, 0.1, 0.2, 0.3, 0.4, 0.5 and 0.6 mM SA was evaluated on seed germination, growth and yield of sweet pepper cv. Yolo wonder at salinity stress on 60 mM NaCl. Seeds were primed with SA concentrations and incubated till 312 h in an incubator to study germination. Same SA concentrations were sprayed on foliage of plants grown in saline soil (60 mM NaCl).ResultsSeeds primed by 0.2 to 0.3 mM SA improved germination rate by 33% due to suppression of ethylene from 3.19 (control) to 2.23–2.70 mg plate−1. Electrolyte leakage reduced to 20.8–21.3% in seeds treated by 0.2–0.3 mM SA compared to 39.9% in untreated seeds. Results also explored that seed priming by 0.3 mM improved TSS, SOD and chlorophyll contents from 13.7 to 15.0 mg g−1 FW, 4.64 to 5.38 activity h−1 100 mg−1 and 89 to 102 ug g−1 compared to untreated seeds, respectively. Results also explore that SA up to 0.2 mM SA applied on plant foliage improved LAI (5–13%), photosynthesis (4–27%), WUE (11–57%), dry weight (5–20%), SOD activity (4–20%) and finally fruit yield (4–20%) compared to untreated plants by ameliorating effect of 60 mM NaCl. Foliar application of SA also caused significant increase in nutrient use efficiency due to significant variations in POD and SOD activities.ConclusionSalicylic acid suppressed ethylene evolution from germinating seeds up to 30% under stress of 60 mM NaCl due to elevated levels of TSS and SOD activity. Foliar application of SA upgraded SOD by lowering POD activity to improve NUE particularly K use efficiency at salinity stress of 60 mM NaCl. Application of 0.2 and 0.3 mM SA emerged as the most effective concentrations of SA for mitigating 60 mM NaCl stress on different physiological and morphological characteristics of sweet pepper.

Chemical Modification of 1-Aminocyclopropane Carboxylic Acid (ACC) Oxidase: Cysteine Mutational Analysis, Characterization, and Bioconjugation with a Nitroxide Spin Label.

1-Aminocyclopropane carboxylic acid oxidase (ACCO) catalyzes the last step of ethylene biosynthesis in plants. Although some sets of structures have been described, there are remaining questions on the active conformation of ACCO and in particular, on the conformation and potential flexibility of the C-terminal part of the enzyme. Several techniques based on the introduction of a probe through chemical modification of amino acid residues have been developed for determining the conformation and dynamics of proteins. Cysteine residues are recognized as convenient targets for selective chemical modification of proteins, thanks to their relatively low abundance in protein sequences and to their well-mastered chemical reactivity. ACCOs have generally 3 or 4 cysteine residues in their sequences. By a combination of approaches including directed mutagenesis, activity screening on cell extracts, biophysical and biochemical characterization of purified enzymes, we evaluated the effect of native cysteine replacement and that of insertion of cysteines on the C-terminal part in tomato ACCO. Moreover, we have chosen to use paramagnetic labels targeting cysteine residues to monitor potential conformational changes by electron paramagnetic resonance (EPR). Given the level of conservation of the cysteines in ACCO from different plants, this work provides an essential basis for the use of cysteine as probe-anchoring residues.

Inducing salt tolerance in wheat through inoculation with rhizobacteria containing 1-aminocyclopropane-1-carboxylate deaminase activity

Wheat (Triticum aestivum) is the fundamental feed of people as it contributes 60% of the daily diet of ordinary man in Pakistan. Salinity is one of the most imperative stresses that hinder agricultural productivity in nearly every part of the world. Improved biosynthesis of ethylene in plants under salinity stress is well established. Higher ethylene concentration retards root growth and eventually disturbs the overall plant growth. Plant growth promoting Rhizobacteria (PGPR) emits 1-aminocyclopropane-1-carboxylate (ACC) deaminase action under salt stressed conditions which minimizes the power of ACC and ethylene justifying the lethal effects of salt stress on plant growth. The seeds inoculated with PGPR having ACC deaminase are comparatively more tolerant to salt stress. The study was conducted at National Agriculture Research Centre Islamabad to examine the influence of PGPR on wheat growth (cultivars Pak-13 NARC-11 and NARC-09) and ionic concentration under saline environment to see the impact of bacterial strains having ACC deaminase on wheat growth and ionic concentration. The experiment was set up following completely randomized design with three repeats. Wheat seeds were inoculated following rhizobacteria strains, WPR-61, WPR-51, WPS-09 and Consortium of WPR-61, WPR-51 and WPS-09. Salinity (10.62dS m-1) was artificially developed using salts. Shoot and root length significantly affected by different rhizobial strains. The maximum root and shoot length attained by the consortium of three strains. The best results were achieved on NARC-09 wheat variety.Pak-13 wheat variety was significant affected by different rhizobial strains and maximum phosphorus concentration attained by WPS-09 strain in Pak-13.

Characterization of Cu(II)-reconstituted ACC Oxidase using experimental and theoretical approaches

1-Aminocyclopropane-1-carboxylic acid oxidase (ACCO) is a non heme iron(II) containing enzyme that catalyzes the final step of the ethylene biosynthesis in plants. The iron(II) ion is bound in a facial triad composed of two histidines and one aspartate (H177, D179 and H234). Several active site variants were generated to provide alternate binding motifs and the enzymes were reconstituted with copper(II). Continuous wave (cw) and pulsed Electron Paramagnetic Resonance (EPR) spectroscopies as well as Density Functional Theory (DFT) calculations were performed and models for the copper(II) binding sites were deduced. In all investigated enzymes, the copper ion is equatorially coordinated by the two histidine residues (H177 and H234) and probably two water molecules. The copper-containing enzymes are inactive, even when hydrogen peroxide is used in peroxide shunt approach. EPR experiments and DFT calculations were undertaken to investigate substrate's (ACC) binding on the copper ion and the results were used to rationalize the lack of copper-mediated activity.

Detection and Amplification of the E8 Promoter in Tomato Plant and Sequencing of the E8 Gene

Tomato (Lycopersicon esculentum Mill) is edible, often red berry-type fruit and a rich source of Fe, Vitamin A, B and C. It is widely cultivated all over the world. E8 gene is related to ethylene biosynthesis in plants. Ethylene is responsible for growth and ripening of fruits. The present study was carried out to detect and amplify the E8 Promoter in the leaves and fruits of Tomato, Spring onion, Capsicum and Lettuce. The CTAB Protocol of Doyle and Doyle (1990) were used for DNA extraction from leaves and fruits and quality was checked on gel electrophoresis. PCR program for E8 was optimized and PCR products were sequenced. The sequencing results were aligned and Blast. The result indicated that in Tomato fruit the E8 gene is detected and amplified as compare to leaves and other plants. This E8 primer will be utilized for different fruits as well.

Detection and Amplification of the E8 Promoter in Tomato Plant and Sequencing of the E8 Gene

Tomato (Lycopersicon esculentum Mill) is edible, often red berry-type fruit and a rich source of Fe, Vitamin A, B and C. It is widely cultivated all over the world. E8 gene is related to ethylene biosynthesis in plants. Ethylene is responsible for growth and ripening of fruits. The present study was carried out to detect and amplify the E8 Promoter in the leaves and fruits of Tomato, Spring onion, Capsicum and Lettuce. The CTAB Protocol of Doyle and Doyle (1990) were used for DNA extraction from leaves and fruits and quality was checked on gel electrophoresis. PCR program for E8 was optimized and PCR products were sequenced. The sequencing results were aligned and Blast. The result indicated that in Tomato fruit the E8 gene is detected and amplified as compare to leaves and other plants. This E8 primer will be utilized for different fruits as well.

Apple MdACS6 Regulates Ethylene Biosynthesis During Fruit Development Involving Ethylene-Responsive Factor.

Ethylene biosynthesis in plants involves different 1-aminocyclopropane-1-carboxylic acid synthase (ACS) genes. The regulation of each ACS gene during fruit development is unclear. Here, we characterized another apple (Malus×domestica) ACS gene, MdACS6. The transcript of MdACS6 was observed not only in fruits but also in other tissues. During fruit development, MdACS6 was initiated at a much earlier stage, whereas MdACS3a and MdACS1 began to be expressed at 35 d before harvest and immediateley after harvest, respectively. Moreover, the enzyme activity of MdACS6 was significantly lower than that of MdACS3a and MdACS1, accounting for the low ethylene biosynthesis in young fruits. Overexpression of MdACS6 (MdACS6-OE) by transient assay in apple showed enhanced ethylene production, and MdACS3a was induced in MdACS6-OE fruits but not in control fruits. In MdACS6 apple fruits silenced by the virus-induced gene silencing (VIGS) system (MdACS6-AN), neither ethylene production nor MdACS3a transcript was detectable. In order to explore the mechanism through which MdACS3a was induced in MdACS6-OE fruits, we investigated the expression of apple ethylene-responsive factor (ERF) genes. The results showed that the expression of MdERF2 was induced in MdACS6-OE fruits and inhibited in MdACS6-AN fruits. Yeast one-hybrid assay showed that MdERF2 protein could bind to the promoter of MdACS3a. Moreover, down-regulation of MdERF2 in apple flesh callus led to a decrease of MdACS3a expression, demonstrating the regulation of MdERF2 on MdACS3a. The mechanism through which MdACS6 regulates the action of MdACS3a was discussed.

History of Research on the Plant Hormone Ethylene

Ethylene is the simplest of the olefin gasses and was the first known gaseous biological signaling molecule. It is synthesized by plants during certain stages of development and in response to abiotic and biotic stresses. Ethylene affects many aspects of plant growth, development as well as responses to environmental cues. Research leading to the discovery of ethylene as a plant hormone started in the 1800s with scientists examining the effects of illuminating gas on plants. In 1901, Dimitry Neljubow determined that ethylene is the active component of illuminating gas that affects plants and thus launched this important field of research. It is generally accepted that in 1934 Richard Gane provided the conclusive evidence that plants biosynthesize ethylene. This early research showed that ethylene is both biosynthesized and sensed by plants. From the 1930s to the 1960s, there was scant research on ethylene as a hormone because many researchers did not believe that ethylene was indeed a plant hormone and because that the detection of ethylene was difficult. However, in the late 1950s, the application of gas chromatography led to an increased interest in ethylene research. From the 1960s through the early 1980s, the biochemical pathway for ethylene biosynthesis in plants was elucidated and membrane-bound ethylene-binding sites were discovered and characterized. The use of Arabidopsis thaliana as a model plant system and the widespread use of molecular biological techniques starting in the 1980s correlates with a second and larger increase in ethylene research productivity. Information gleaned from this model plant is now being applied to many plant species. In recent years, detailed models for the regulation of ethylene biosynthesis and ethylene signal transduction have emerged. This article provides an overview of the key historical discoveries regarding ethylene as a plant hormone.

Real time expression of ACC oxidase and PR-protein genes mediated by Methylobacterium spp. in tomato plants challenged with Xanthomonas campestris pv. vesicatoria

Biotic stress like pathogenic infection increases ethylene biosynthesis in plants and ethylene inhibitors are known to alleviate the severity of plant disease incidence. This study aimed to reduce the bacterial spot disease incidence in tomato plants caused by Xanthomonas campestris pv. vesicatoria (XCV) by modulating stress ethylene with 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity of Methylobacterium strains. Under greenhouse condition, Methylobacterium strains inoculated and pathogen challenged tomato plants had low ethylene emission compared to pathogen infected ones. ACC accumulation and ACC oxidase (ACO) activity with ACO related gene expression increased in XCV infected tomato plants over Methylobacterium strains inoculated plants. Among the Methylobacterium spp., CBMB12 resulted lowest ACO related gene expression (1.46 Normalized Fold Expression), whereas CBMB20 had high gene expression (3.42 Normalized Fold Expression) in pathogen challenged tomato. But a significant increase in ACO gene expression (7.09 Normalized Fold Expression) was observed in the bacterial pathogen infected plants. In contrast, Methylobacterium strains enhanced β-1,3-glucanase and phenylalanine ammonia-lyase (PAL) enzyme activities in pathogen challenged tomato plants. The respective increase in β-1,3-glucanase related gene expressions due to CBMB12, CBMB15, and CBMB20 strains were 66.3, 25.5 and 10.4% higher over pathogen infected plants. Similarly, PAL gene expression was high with 0.67 and 0.30 Normalized Fold Expression, in pathogen challenged tomato plants inoculated with CBMB12 and CBMB15 strains. The results suggest that ethylene is a crucial factor in bacterial spot disease incidence and that methylobacteria with ACC deaminase activity can reduce the disease severity with ultimate pathogenesis-related protein increase in tomato.

A Simple Complex on the Verge of Breakdown: Isolation of the Elusive Cyanoformate Ion

Why does cyanide not react destructively with the proximal iron center at the active site of 1-aminocyclopropane-1-carboxylic acid (ACC) oxidase, an enzyme central to the biosynthesis of ethylene in plants? It has long been postulated that the cyanoformate anion, [NCCO2](-), forms and then decomposes to carbon dioxide and cyanide during this process. We have now isolated and crystallographically characterized this elusive anion as its tetraphenylphosphonium salt. Theoretical calculations show that cyanoformate has a very weak C-C bond and that it is thermodynamically stable only in low dielectric media. Solution stability studies have substantiated the latter result. We propose that cyanoformate shuttles the potentially toxic cyanide away from the low dielectric active site of ACC oxidase before breaking down in the higher dielectric medium of the cell.

Microbial ACC-Deaminase: Prospects and Applications for Inducing Salt Tolerance in Plants

Salinity is one of the most important stresses that hamper agricultural productivity in nearly every part of the world. Enhanced biosynthesis of ethylene in plants under salinity stress is well established. Higher ethylene concentration inhibits root growth and ultimately affects the overall plant growth. Overcoming this ethylene-induced root inhibition is a prerequisite for successful crop production. Recent studies have shown that ethylene level in plants is regulated by a key enzyme 1-aminocyclopropane-1-carboxylicacid (ACC)-deaminase. This enzyme is present in plant growth-promoting bacteria (PGPR) and lowers the ethylene level by metabolizing its precursor ACC into α-ketobutyrate and ammonia (NH3). Inoculation of plants under salinity stress with PGPR having ACC-deaminase activity mitigates the inhibitory effects of salinity on root growth by lowering the ethylene concentration in the plant. This in turn results in prolific root growth, which is beneficial for the uptake of nutrients and maintenance of growth under stressful environment. The present review critically discusses the effects of salinity stress on plant growth with special reference to ethylene production and the effects of rhizobacteria containing ACC-deaminase on crop improvement under salinity stress. It also discusses how much progress has been made in producing transgenic lines of different crops over-expressing the gene encoding ACC-deaminase and how far such transformed lines can tolerate salinity stress.

Molecular evolution of the E8 promoter in tomato and some of its relative wild species

The E8 gene is related to ethylene biosynthesis in plants. To explore the effect of the expression pattern of the E8 gene on different E8 promoters, the molecular evolution of E8 promoters was investigated. A total of 16 E8 promoters were cloned from 16 accessions of seven tomato species,and were further analysed. The results from 19 E8 promoters including three previously cloned E8 promoters (X13437,DQ317599 and AF515784) showed that the size of the E8 promoters varied from 2101 bp (LA2150) to 2256 bp (LA2192); their sequences shared 69.9% homology and the average A/T content was 74.9%. Slide-window analysis divided E8 promoters into three regions -A,B and C - and the sequence identity in these regions was 72.5%, 41.2% and 70.8%, respectively. By searching the cis -elements of E8 promoters in the PLACE database, mutant nucleotides were found in some functional elements,and deletions or insertions were also found in regions responsible for ethylene biosysnthesis (-1702 to -1274) and the negative effect region (-1253 to -936). Our results indicate that the size of the functional region for ethylene biosynthesis in the E8 promoter could be shortened from 429 bp to 113 bp (-1612 to -1500). The results of molecular evolution analysis showed that the 19 E8 promoters could be classified into four clade groups, which is basically consistent with evolution of the tomato genome. Southern blot analysis results showed that the copy number of E8 promoters in tomato and some other wild species changed from 1 to 4. Taken together, our study provides important information for further elucidating the E8 gene expression pattern in tomato, analysing functional elements in the E8 promoter and reconstructing the potent E8 promoter.

Quantum Chemical Study on Stability and Reactivity of 1-Aminocyclopropane-1-carboxylic Acid Amine Radical Cation

Abstract The stability and reactivity of the 1-aminocyclopropane-1-carboxylic acid (ACC) amine radical cation, which is a key intermediate of ethylene biosynthesis in plants, were examined by quantum chemical calculations. The local interactions to stabilize radical species were treated with a hydrated cluster. Potential energy curves were obtained for two types of reactions from ACC amine radical cation: (1) direct ring opening of ACC amine radical cation, followed by proton abstraction from the amino group, and (2) proton abstraction from the amino group, followed by ring opening of ACC aminyl radical. A remarkable difference was found in the ring-opening processes of reactions (1) and (2). Rate constants (k) of 6.10×1012 and 8.89×109 s−1 at 298.15 K were estimated for the ring openings of ACC amine radical cation and ACC aminyl radical, respectively, at the B3LYP/6-31+G(d,p) level, and were in agreement with experimental results. The ring opening of ACC amine radical cation was 103-fold faster than that of ACC aminyl radical with almost no barrier height. It was thus quantitatively demonstrated that reaction (1) was more favorable than reaction (2) for ethylene synthesis.

Rhizobitoxine production inAgrobacterium tumefaciensC58 byBradyrhizobium elkanii rtxACDEFGgenes

We examined the genetic basis and transfer for production of rhizobitoxine, an inhibitor of ethylene biosynthesis in plants, directed by the rtx genes of Bradyrhizobium elkanii. Comparison with genome sequences of Bradyrhizobium japonicum and Xanthomonas oryzae suggests that the rtx genes extend from the previously identified rtxAC genes through four additional genes rtxDEFG. Reverse transcription-PCR analysis showed that the rtxACDEFG genes are expressed as an operon. Mutational analysis indicated that rtxDEG mutants reduced rhizobitoxine biosynthesis, while the rtxA gene is essential for its synthesis. Introduction of the rtxACDEFG into Agrobacterium tumefaciens resulted in strong expression of rtxACDEFG and production of RtxA protein, but no rhizobitoxine was detectable. Addition of O-acetylhomoserine, a precursor of rhizobitoxine, to the Agrobacterium derivative, however, fostered production of rhizobitoxine in culture. The diluted culture supernatant inhibited the activities of beta-cystathionase and 1-aminocyclopropane-1-carboxylate synthase, indicating that A. tumefaciens carrying rtxACDEFG genes excreted biologically active rhizobitoxine.

Effect of salt and osmotic stress upon expression of the ethylene receptor ETR1 in Arabidopsis thaliana

In hormone perception, varying the concentrations of hormone, receptor, or downstream signaling elements can modulate signal transduction. Previous research has demonstrated that ethylene biosynthesis in plants is regulated by abiotic factors. Here we report that exposure of Arabidopsis plants to NaCl reduced expression of the ethylene receptor ETR1. The change in gene expression was reflected at the protein level based on immunoblot analysis. Further analysis supports a general effect of osmotic stress upon the expression level of ETR1. The reduction in ETR1 levels should cause increased sensitivity of the plant to ethylene. These results suggest that plant responses to abiotic stress are modulated by changes in the expression level of ethylene receptors.

Activation of a stress-responsive mitogen-activated protein kinase cascade induces the biosynthesis of ethylene in plants.

Plants under stress from both biotic and abiotic sources produce increased levels of ethylene, which is perceived by ethylene receptors and triggers cellular responses further downstream. Protein phosphorylation and dephosphorylation were implicated in the regulation of ethylene induction by stresses based on studies using protein kinase and phosphatase inhibitors. However, the kinase(s) involved remains to be determined. Using a conditional gain-of-function transgenic system, we demonstrate that the activation of SIPK, a tobacco mitogen-activated protein kinase (MAPK), by NtMEK2DD, an active mutant of the upstream kinase of SIPK, resulted in a dramatic increase in ethylene production. The increase in ethylene after the activation of SIPK coincided with a dramatic increase in 1-aminocyclopropane-1-carboxylic acid (ACC) synthase (ACS) activity, which was followed by the activation of a subgroup of ACS and ACC oxidase (ACO) genes, suggesting that either the activation of unidentified ACS(s) or post-transcriptional regulation is involved. Infection with Tobacco mosaic virus (TMV), which is known to activate the SIPK cascade and induce ethylene biosynthesis, also induced the same ACSs and ACOs. After ethylene production in NtMEK2DD plants, strong activation of ETHYLENE-RESPONSE FACTOR (ERF) genes was observed, similar to the effect in NN tobacco plants infected with TMV. In contrast to previous reports, no major increase in jasmonic acid (JA) and methyl jasmonate (MJ) was detected after the activation of SIPK/WIPK in NtMEK2DD transgenic plants. These results suggest that the induction of ethylene but not JA/MJ is involved in plant defense responses mediated by the NtMEK2-SIPK/WIPK pathway.