Stress-induced Ethylene Production Research Articles

Ethylene inhibitors improve crop productivity by modulating gene expression, antioxidant defense machinery and photosynthetic efficiency of Solanum lycopersicum L. cv. Pusa Ruby grown in controlled salinity stress conditions

Tomato is an important horticultural crop of global importance but its susceptibility to salinity stress significantly retards growth and thereby its yield. The impact of gaseous hormone, ethylene, accentuates the effect of salinity stress by negatively regulating various growth and developmental processes. Ethylene overproduction and ionic imbalance triggered by salinity stress exhibit upregulation of 1-aminocyclopropane-1-carboxylic acid synthase (ACS) and downregulation of NHX1, SOS1 and HKT1;2 under salinity stress conditions that negatively affect proline content, ROS scavenging machinery, morphometric and physiological attributes. Variation in the proline content, activation of ROS scavenging machinery, affected morphometric and physiological attributes are some discernible points in response to salinity stress induced ethylene production. Therefore, the experiment was designed to study the role of ethylene inhibitors silver nitrate (AgNO3) and cobalt chloride (CoCl2) in mitigating the negative effects of ethylene over production under salinity stress. Tomato seedlings were treated with 0 mM NaCl, 250 mM NaCl and 250 mM NaCl + 100 µM AgNO3, 250 mM NaCl + 100 µM CoCl2 for three days. The gene expression of ACS2, NHX1, SOS1 and HKT1;2, and ACS activity, endogenous ethylene level, Na+ and K+ ions concentration, proline content, antioxidant enzymes activity, photosynthetic efficiency and different morphological and yield parameters were assessed in tomato plants treated with salinity stress, and ethylene inhibitors at 45 DAS and 90 DAS. Results revealed that ethylene inhibitors might be playing the role in minimizing the ethylene and ACC load. The ACS enzyme activity and ethylene content were considerably decreased due to application of AgNO3and CoCl2. Further, the endogenous proline and K+ content, and K+/Na+ ratio were considerably increased in response to ethylene inhibitors treatment. Also, antioxidant enzymesviz., SOD, CAT, APX and GR, growth matrices, photosynthetic efficiency and yield attributes were significantly improved in treated tomato plants (salinity with ethylene inhibitors).

Stress-induced ethylene production by soybean symbiotic systems depending on genetic characterization of Bradyrhizobium strains

Stress-induced ethylene production by soybean symbiotic systems depending on genetic characterization of Bradyrhizobium strains

Overexpression of 1-Aminocyclopropane-1-Carboxylic Acid Deaminase (acdS) Gene in Petunia hybrida Improves Tolerance to Abiotic Stresses.

Abiotic stress induces the ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC) in plants, which consequently enhances ethylene production and inhibits plant growth. The bacterial ACC deaminase enzyme encoded by the acdS gene reduces stress-induced ethylene production and improves plant growth in response to stress. In this study, overexpression of acdS in Petunia hybrida (‘Mirage Rose’) significantly reduced expression of the ethylene biosynthesis gene ACC oxidase 1 (ACO1) and ethylene production relative to those in wild type (WT) under various abiotic stresses (cold, drought, and salt). The higher reduction of stress-induced ethylene in the transgenic plants, which was due to the overexpression of acdS, led to a greater tolerance to the stresses compared to that in the WT plants. The greater stress tolerances were proven based on better plant growth and physiological performance, which were linked to stress tolerance. Moreover, expression analysis of the genes involved in stress tolerance also supported the increased tolerance of transgenics relative to that with the WT. These results suggest the possibility that acdS is overexpressed in ornamental plants, particularly in bedding plants normally growing outside the environment, to overcome the deleterious effect of ethylene on plant growth under different abiotic stresses. The development of stress-tolerant plants will be helpful to advance the floricultural industry.

New insights into the effects of ethylene on ABA catabolism, sweetening and dormancy in stored potato tubers.

Continuous ethylene supplementation suppresses postharvest sprouting, but it can increase reducing sugars, limiting its use as an alternative to chlorpropham for processing potatoes. To elucidate the mechanisms involved, tubers were treated after curing with or without the ethylene binding inhibitor 1-methylcyclopropene (1-MCP at 1 μL L-1 for 24 h), and then stored in air or air supplemented with continuous ethylene (10 μL L-1). Across three consecutive seasons, changes in tuber physiology were assessed alongside transcriptomic and metabolomic analysis. Exogenous ethylene alone consistently induced a respiratory rise and the accumulation of undesirable reducing sugars. The transient respiratory peak was preceded by the strong upregulation of two genes encoding 1-aminocyclopropane-1-carboxylate oxidase (ACO), typical of wound and stress induced ethylene production. Profiles of parenchymatic tissue highlighted that ethylene triggered abscisic acid (ABA) catabolism, evidenced by a steep fall in ABA levels and a transient rise in the catabolite phaseic acid, accompanied by upregulation of transcripts encoding an ABA 8'-hydroxylase. Moreover, analysis of non-structural carbohydrate-related genes revealed that ethylene strongly downregulated the expression of the Kunitz-type invertase inhibitor, already known to be involved in cold-induced sweetening. All these ethylene-induced effects were negated by 1-MCP with one notable exception: 1-MCP enhanced the sprout suppressing effect of ethylene whilst preventing ethylene-induced sweetening. This study supports the conclusions that: i) tubers adapt to ethylene by regulating conserved pathways (e.g. ABA catabolism); ii) ethylene-induced sweetening acts independently from sprout suppression, and is similar to cold-induced sugar accumulation.

Effect of halotolerant rhizobacteria isolated from halophytes on the growth of sugar beet (Beta vulgaris L.) under salt stress.

Utilization of rhizobacteria that have associated with plant roots in harsh environments could be a feasible strategy to deal with limits to agricultural production caused by soil salinity. Halophytes occur naturally in high-salt environments, and their roots may be associated with promising microbial candidates for promoting growth and salt tolerance in crops. This study aimed to isolate efficient halotolerant plant-growth-promoting rhizobacterial strains from halophytes and evaluate their activity and effects on sugar beet (Beta vulgaris L.) growth under salinity stress. A total of 23 isolates were initially screened for their ability to secrete 1-aminocyclopropane-1-carboxylate deaminase (ACD) as well as other plant-growth-promoting characteristics and subsequently identified by sequencing the 16S rRNA gene. Three isolates, identified as Micrococcus yunnanensis, Planococcus rifietoensis and Variovorax paradoxus, enhanced salt stress tolerance remarkably in sugar beet, resulting in greater seed germination and plant biomass, higher photosynthetic capacity and lower stress-induced ethylene production at different NaCl concentrations (50-125 mM). These results demonstrate that salinity-adapted, ACD-producing bacteria isolated from halophytes could promote sugar beet growth under saline stress conditions.

Arabidopsis Protein Phosphatase 2C ABI1 Interacts with Type I ACC Synthases and Is Involved in the Regulation of Ozone-Induced Ethylene Biosynthesis

Ethylene plays a crucial role in various biological processes and therefore its biosynthesis is strictly regulated by multiple mechanisms. Posttranslational regulation, which is pivotal in controlling ethylene biosynthesis, impacts 1-aminocyclopropane 1-carboxylate synthase (ACS) protein stability via the complex interplay of specific factors. Here, we show that the Arabidopsis thaliana protein phosphatase type 2C, ABI1, a negative regulator of abscisic acid signaling, is involved in the regulation of ethylene biosynthesis under oxidative stress conditions. We found that ABI1 interacts with ACS6 and dephosphorylates its C-terminal fragment, a target of the stress-responsive mitogen-activated protein kinase, MPK6. In addition, ABI1 controls MPK6 activity directly and by this means also affects the ACS6 phosphorylation level. Consistently with this, ozone-induced ethylene production was significantly higher in an ABI1 knockout strain (abi1td) than in wild-type plants. Importantly, an increase in stress-induced ethylene production in the abi1td mutant was compensated by a higher ascorbate redox state and elevated antioxidant activities. Overall, the results of this study provide evidence that ABI1 restricts ethylene synthesis by affecting the activity of ACS6. The ABI1 contribution to stress phenotype underpins its role in the interplay between the abscisic acid (ABA) and ethylene signaling pathways.

Isolation and characterization of drought-tolerant ACC deaminase and exopolysaccharide-producing fluorescent Pseudomonas sp.

The enzyme 1-aminocyclopropane-1-carboxylate deaminase catalyzes the degradation of 1-aminocyclopropane-1-carboxylic acid (ACC), the immediate precursor of the plant hormone ethylene, into α-ketobutyrate and ammonia. The enzyme has been detected in a limited number of bacteria and plays a significant role in sustaining plant growth and development under biotic and abiotic stress conditions by reducing stress-induced ethylene production in plants. We have screened 32 fluorescent Pseudomonas sp. isolated from rhizosphere and non-rhizosphere soils of different crop production systems for drought tolerance using polyethylene glycol 6000 (PEG 6000). Nine of these isolates were tolerant to a substrate metric potential of −0.30 MPa (15 % PEG 6000) and therefore considered to be drought-tolerant. All of these drought-tolerant isolates were screened for ACC deaminase activity using ACC as the sole nitrogen source, and one (SorgP4) was found to be positive for ACC, producing 3.71 ± 0.025 and 1.42 ± 0.039 μM/mg protein/h of α-ketobutyrate under the non-stress and drought stress condition, respectively. The isolate SorgP4 also showed other plant growth-promoting traits, such as indole acetic acid production, phosphate solubilization, siderophore and hydrogen cyanide production. The ACC deaminase gene (acdS) from the isolate SorgP4 was amplified, and the nucleotide sequence alignment of the acdS gene showed significant homology with acdS genes of NCBI Genbank. The 16S rRNA gene sequencing analysis identified the isolate as Pseudomonas fluorescens. Both sequences have been submitted to the NCBI GenBank under the accession numbers JX885767 and KC192771 respectively.

스트레스에 의한 식물세포 손상에서 Biphasic Reactive Oxygen Species(ROS)와 Ethylene 생합성의 Synergism 효과

비생물학적 스트레스로 <TEX>$H_2O_2$</TEX>를 이용하여 산화적 스트레스와 고염분 스트레스를 처리한 후 ROS의 생성을 확인한 결과 스트레스 처리 후 30분에 일시적으로 1차 peak를 형성하였다가 거의 basal level까지 감소하고 다시 증가하여 3시간에 매우 다량의 2차 peak를 형성한 후 거의 basal level로 다시 낮아지는 biphasic 양상을 나타내게 된다. 따라서 ROS의 생성은 초기 30분 내에 일시적으로 발생한 Phase I의 ROS와 Phase II의 좀 더 장기적으로 다량의 고농도로 발생된 ROS의 생리적 역할이 다를 것으로 여겨진다. 본 논문에서는 스트레스 처리 시 생성되는 ROS를 확인한 후 ROS 생성 유전자인 RbohD와 RbohF 유전자 발현이 억제된 RbohD-AS, RbohF-AS 형질전환 식물체를 이용하여 실험을 수행하였다. 스트레스에 의해 생성되는 ROS의 생성을 억제시킴으로써 스트레스에 대한 ethylene 생성이 더 적은 것으로 나타났다. 또한, 이들 형질전환 식물체에서 ethylene 생성과 <TEX>$H_2O_2$</TEX> 억제 효과를 확인하였으며 고염분 등의 스트레스에 대한 저항성은 ROS와 ethylene의 생성이 저하되어 나타난 것으로 판단된다. 산화적 스트레스와 고염분 스트레스에서 후기 ethylene이 다량으로 생성되는 시기, 즉 세포손상이 초래되는 후기에서 DNA fragmentation 분석을 통해서 ROS와 ethylene의 생성이 높은 식물체일수록 PCD가 높게 나타난 것으로 여겨지며, 이 과정에서 작용하는 유전자는 RbohD와 RbohF인 것으로 보이며, RbohD가 더 효과적으로 작용하는 것으로 생각된다. 따라서 스트레스에 반응하는 신호전달과정에서 초기에 ROS가 생성이 되고 후기에 ethylene이 다량으로 생성되어 결국 세포죽음에 이르게 하는 상호 synergism을 일으켜 반응을 나타내며, 이러한 반응 과정에서 RbohD와 RbohF 유전자 발현의 억제가 스트레스에 대한 식물체의 저항성을 높이는 것으로 사료된다. Although reactive oxygen species (ROS) are inevitable by-products of many redox reactions in eukaryotic cells, they play a crucial role as signaling molecules in many cellular processes for development and defense response to abiotic stresses. The biphasic ROS production which was peaked twice in a first transient phase and a second massive phase was occurred after treatment of abiotic stress such as oxidative stress, high salinity. This biphasic generation of ROS was followed by the biphasic production of stress hormone, ethylene. The mechanism of interactions between ROS and ethylene biosynthesis is studied in tobacco (Nicotiana tabaccum L.) plants under the abiotic stresses. The stress-induced ethylene production was significantly inhibited in RbohD-AS and RbohF-AS, in which antisense expression of NADPH oxidase genes was performed. The accumulation of ROS, which was determined by DAB and DCFH-DA staining, was significantly decreased after abiotic stresses in transgenic plants. The suppression of signaling with ethylene and ROS induced more tolerance in response to abiotic stress. The transgenic plants were more tolerant in MS medium supplemented with salinity stress in contrast with wild-type. Stress-induced cell damage determined by DNA fragmentation was decreased at phase II in those transgenic plants. Therefore, the first burst of ROS is more responsible for making a role as a signaling molecule during stress-induced response. These results suggested that ethylene and ROS act in a positive feedback cycle that results in mutual enhancement of ethylene and ROS production during stress-induced cell death.

Enhancement of growth and salt tolerance of red pepper seedlings (Capsicum annuum L.) by regulating stress ethylene synthesis with halotolerant bacteria containing 1-aminocyclopropane-1-carboxylic acid deaminase activity

Three 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase-producing halotolerant bacteria were isolated from West Coast soil of Yellow Sea, Incheon, South Korea and evaluated for their efficiency in improving red pepper plant growth under salt stress. The strains RS16, RS656 and RS111 were identified by 16S rRNA gene sequencing as Brevibacterium iodinum, Bacillus licheniformis and Zhihengliuela alba, respectively. Two hour exposure of 100, 150 and 200 mM NaCl stress on 8 day old red pepper seedlings caused 44, 64 and 74% increase ethylene production, while at 150 mM NaCl stress, inoculation of B. licheniformis RS656, Z. alba RS111, and Br. iodinum RS16 reduces ethylene production by 44, 53 and 57%, respectively. Similarly, 3 week old red pepper plants were subjected to salt stress for two weeks and approximately ∼50% reduction in growth recorded at 150 mM NaCl stress compared to negative control whereas bacteria inoculation significantly increase the growth compared to positive control. Salt stress also caused 1.3-fold reduction in the root/shoot dry weight ratio compared to the absence of salt while bacteria inoculation retained the biomass allocation similar to control plants. The salt tolerance index (ratio of biomass of salt stressed to non-stressed plant) was also significantly increased in inoculated plants compared to non-inoculated. Increase nutrient uptakes under salt stress by red pepper further evident that bacteria inoculation ameliorates salt stress effect. In summary, this study indicates that the use of ACC deaminase-producing halotolerant bacteria mitigates the salt stress by reducing salt stress-induced ethylene production on growth of red pepper plants.

Biochemical and Molecular Characterization of Mesorhizobium ciceri Containing acdS Gene

The enzyme ACC deaminase encoded by acdS gene is known to play a significant role in sustaining plant growth and development under stress conditions by reducing stress induced ethylene production inside the plant. In the present investigation, Mesorhizobium ciceri isolates from chickpea plants growing under drought conditions were screened for the presence of acdS gene. Full length acdS gene from one of the isolates was successfully amplified. This is the first report showing presence of acdS gene in Mesorhizobium ciceri. Gene sequence analysis revealed that there were deletions at two positions in this gene as compared to M. loti acdS gene.

Perspective of plant growth promoting rhizobacteria (PGPR) containing ACC deaminase in stress agriculture

Ethylene is a gaseous plant growth hormone produced endogenously by almost all plants. It is also produced in soil through a variety of biotic and abiotic mechanisms, and plays a key role in inducing multifarious physiological changes in plants at molecular level. Apart from being a plant growth regulator, ethylene has also been established as a stress hormone. Under stress conditions like those generated by salinity, drought, waterlogging, heavy metals and pathogenicity, the endogenous production of ethylene is accelerated substantially which adversely affects the root growth and consequently the growth of the plant as a whole. Certain plant growth promoting rhizobacteria (PGPR) contain a vital enzyme, 1-aminocyclopropane-1-carboxylate (ACC) deaminase, which regulates ethylene production by metabolizing ACC (an immediate precursor of ethylene biosynthesis in higher plants) into alpha-ketobutyrate and ammonia. Inoculation with PGPR containing ACC deaminase activity could be helpful in sustaining plant growth and development under stress conditions by reducing stress-induced ethylene production. Lately, efforts have been made to introduce ACC deaminase genes into plants to regulate ethylene level in the plants for optimum growth, particularly under stressed conditions. In this review, the primary focus is on giving account of all aspects of PGPR containing ACC deaminase regarding alleviation of impact of both biotic and abiotic stresses onto plants and of recent trends in terms of introduction of ACC deaminase genes into plant and microbial species.

Heat stress induced ethylene production in developing wheat grains induces kernel abortion and increased maturation in a susceptible cultivar

In this work we present evidence that the heat stress induced kernel abortion and suppression of grain maturation in a representative heat susceptible hard red winter wheat ( Triticum aestivum L.) cultivar is regulated by heat stress induced ethylene production. Exposure to heat stress (38 °C) during early kernel development (10 DAP) resulted in a 6-fold increase in ethylene production in developing kernels of the heat susceptible hard red winter wheat cultivar ‘Karl 92’. A similar 7-fold increase in ethylene production in embryos and 12-fold increase in ethylene production in the flag leaf of heat stressed plants of ‘Karl 92’ was also found. In contrast, no change in ethylene production was observed in the heat tolerant hard white spring wheat cultivar ‘Halberd’. In an effort to link the heat stress induced ethylene production to the observed increase in kernel abortion and reduced kernel weight in the heat susceptible ‘Karl 92’, plants were treated with the ethylene receptor inhibitor 1-methylcyclopropane (1-MCP) prior to exposure to heat stress. Inhibiting ethylene perception in the heat susceptible ‘Karl 92’ in this manner blocked heat stress induced kernel abortion and reduction in kernel weight and demonstrated a clear link between ethylene in regulating susceptibility to heat stress or perception of high temperatures as a timing signal for transitioning to developmental arrest and senescence in certain wheat genotype classes.

Ozone-induced ethylene production is dependent on salicylic acid, and both salicylic acid and ethylene act in concert to regulate ozone-induced cell death.

Ethylene is known to influence plant defense responses including cell death in response to both biotic and abiotic stress factors. However, whether ethylene acts alone or in conjunction with other signaling pathways is not clearly understood. Ethylene overproducer mutants, eto1 and eto3, produced high levels of ethylene and developed necrotic lesions in response to an acute O3 exposure that does not induce lesions in O3-tolerant wild-type Col-0 plants. Treatment of plants with ethylene inhibitors completely blocked O3-induced ethylene production and partially attenuated O3-induced cell death. Analyses of the responses of molecular markers of specific signaling pathways indicated a relationship between salicylic acid (SA)- and ethylene-signaling pathways and O3 sensitivity. Both eto1 and eto3 plants constitutively accumulated threefold higher levels of total SA and exhibited a rapid increase in free SA and ethylene levels prior to lesion formation in response to O3 exposure. SA pre-treatments increased O3 sensitivity of Col-0, suggesting that constitutive high SA levels prime leaf tissue to exhibit increased magnitude of O3-induced cell death. NahG and npr1 plants compromised in SA signaling failed to produce ethylene in response to O3 and other stress factors suggesting that SA is required for stress-induced ethylene production. Furthermore, NahG expression in the dominant eto3 mutant attenuated ethylene-dependent PR4 expression and rescued the O3-induced HR (hypersensitive response) cell death phenotype exhibited by eto3 plants. Our results suggest that both SA and ethylene act in concert to influence cell death in O3-sensitive genotypes, and that O3-induced ethylene production is dependent on SA.

The effect of water stress on 1-(malonylamino)cyclopropane-1-carboxylic acid concentration in plant tissues.

Since the discovery of1-(malonylamino)cyclopropane-1-carboxylic acid (MACC)as a major metabolite of both endogenous andexogenously applied 1-aminocyclopropane-1-carboxylicacid (ACC), it has become evident that the formationof MACC from ACC can act to regulate ethyleneproduction in certain tissues. Hence it was suggestedthat MACC could serve as an indicator of water-stresshistory in plant tissues. The accurate quantificationof MACC in plant tissues is essential forunderstanding the role of MACC in the regulation ofethylene biosynthesis.Hoffman et al. [15] described a method for themeasurement of MACC in which MACC was hydrolysed byHCl to ACC, which was then assayed by chemicaloxidation to form ethylene. Attempts have been made byothers to raise monoclonal antibodies to MACC so thatan immunoassay could be developed in order to gain adeeper understanding of stress-induced ethyleneproduction but no further publications have beenforthcoming.Here a method employing GC-MS is compared with theindirect assay for MACC, which is based uponhydrolysis of MACC to ACC and conversion of ACC byhypochlorite reagent to ethylene which is subsequentlyquantified by GC.

Stress induced ethylene production, ethylene binding, and the response to the ethylene action inhibitor 1-MCP in miniature roses

In an interior environment, the display life of miniature potted roses ( Rosa hybrida L.) ‘Bronze’ was 7 days, while that of ‘Vanilla’ was 23 days. Although there was a substantial difference in ethylene production between flowers from two cultivars, there was no difference in binding site activity ( k d = 0.14 nl l −1). Pre-treatment with the ethylene action inhibitor 1-methylcyclopropene (1-MCP) did not have a clear effect on display life without preceding transport stress. When the plants were subjected to simulated transportation stress, post-harvest life of both cultivars was reduced (to 6 days for ‘Bronze’, and to 17 days for ‘Vanilla’). In ‘Bronze’ the stress resulted in a modest increase in ethylene production, but there was no effect of stress on the low ethylene production by ‘Vanilla’ flowers. Nevertheless, pretreatment with 1-MCP before transport simulation improved display life of both cultivars.

Expression Characteristics of CS-ACS1, CS-ACS2 and CS-ACS3, Three Members of the 1-Aminocyclopropane-1-Carboxylate Synthase Gene Family in Cucumber (Cucumis sativus L.) Fruit under Carbon Dioxide Stress

We investigated the expression pattern of three 1-aminocyclopropane-1-carboxylate (ACC) synthase genes, CS-ACS1, CS-ACS2 and CS-ACS3 in cucumber (Cucumis sativus L.) fruit under CO2 stress. CO2 stress-induced ethylene production paralleled the accumulation of only CS-ACS1 transcripts which disappeared upon withdrawal of CO2. Cycloheximide inhibited the CO2 stress-induced ethylene production but superinduced the accumulation of CS-ACS1 transcript. At higher concentrations, cycloheximide also induced the accumulation of CS-ACS2 and CS-ACS3 transcripts. In the presence of CO2 and cycloheximide, the accumulation of CS-ACS2 transcript occurred within 1 h, disappeared after 3 h and increased greatly upon withdrawal of CO2. Inhibitors of protein kinase and types 1 and 2A protein phosphatases which inhibited and stimulated, respectively, CO2 stress-induced ethylene production had little effect on the expression of these genes. The results presented here identify CS-ACS1 as the main ACC synthase gene responsible for the increased ethylene biosynthesis in cucumber fruit under CO2 stress and suggest that this gene is a primary response gene and its expression is under negative control since it is expressed by treatment with cycloheximide. The results further suggest that the regulation of CO2 stress-induced ethylene biosynthesis by reversible protein phosphorylation does not result from enhanced ACC synthase transcription.

Characterization of Carbon Dioxide Stress-Induced Ethylene Biosynthesis in Cucumber (Cucumis sativus L.) Fruit

In the present study, we have investigated the mechanism through which carbon dioxide induces ethylene biosynthesis in cucumber {Cucumis sativus L.) fruit. A series of inhibitors were tested in order to determine the involvement of de novo protein synthesis, new mRNA synthesis, protein kinase activity, and phosphoprotein phospbatase activity in carbon dioxide stress-induced ethylene biosynthesis. Carbon dioxide stimulated ethylene production through the induction of both 1-aminocyclopropane-l-carboxylate (ACC) synthase and ACC oxidase activities. Induction of ACC synthase activity led to accumulation of ACC. Cycloheximide blocked carbon dioxide stress-induced ethylene biosynthesis while cordycepin had no effect. Dibucaine and 6-dimethylaminopurine, inhibitors of protein kinases, blocked carbon dioxide stress-induced increases in ethylene production, ACC synthase activity, and ACC content but not the increase in ACC oxidase activity. Cantharidin, a potent inhibitor of protein phosphatase type 1 and type 2A, stimulated carbon dioxide stress-induced ethylene production, ACC synthase activity, and ACC accumulation but had no additional effect on carbon dioxide stress-induced ACC oxidase activity. In the presence of dibucaine or cycloheximide, the stimulation of carbon dioxide stress-induced ethylene biosynthesis by cantharidin was blocked. These data suggest that carbon dioxide stress-induced ethylene biosynthesis may be regulated posttranscriptionally and that the induction involves the synthesis of novel protein(s) in the cytosol. The results further identify protein phosphorylation and dephosphorylation as requirements in one or more of the steps involved in the carbon dioxide signal transduction pathway that leads to induction of ethylene biosynthesis and, in particular to the induction of ACC synthase, presumably the key enzyme in the ethylene biosynthetic pathway.

Iron toxicity and stress-induced ethylene production in rice leaves

The relationship among iron toxicity, bronzing symptom, and stress-induced ethylene production (SEP) was investigated in detached rice (Oryza sativa L.) leaves during the vegetative-ripening stage and in whole plants during the vegetative stage. When Fe2+ (200 mg L-1) was applied to the detached leaf through a transpiration stream, SEP was higher in the first leaf than in the second and third leaves from the top and maximal around the panicle primordia initiation stage. The genotype difference in SEP was more pronounced in the second and third leaves than in the first leaf. Bronzing intensity increased as SEP increased; iron concentration increase during treatment in the tissue did not correlate with bronzing intensity or with SEP among the 16 genotypes tested. When the roots of an intact plant were exposed to 300 mg L-1 of Fe2+ in culture solution little stress-induced ethylene was produced. By partially or totally derooting the plant, however, stress-induced ethylene was evoked, indicating that roots reduced the Fe2+ uptake so that little stress ethylene is produced in the intact plant. Leaf tissue tolerance for Fe2+ may contribute to genotype differences in iron toxicity tolerance of rice plants when roots are injured during transplanting or exposed to toxic substances in the soil.

Induction of ethylene biosynthesis and polyamine accumulation in cucumber fruit in response to carbon dioxide stress

Abstract Carbon dioxide stress-induced ethylene biosynthesis, respiration and polyamine accumulation in cucumber fruit ( Cucumis sativus L. cv. Sharp-1) held at 25 °C was investigated. Control fruit produced little ethylene and the respiration rate decreased with increase in incubation time while polyamine levels decreased. Elevated CO 2 induced ethylene production, respiration and polyamine accumulation. Putrescine and spermidine levels increased in response to CO 2 treatment, whereas spermine levels were not significantly affected. No cadaverine was detected in all treatments. The increase in ethylene production paralleled increases in 1-aminocyclopropane-1-carboxylic acid (ACC) and the activities of both ACC synthase and in vitro ACC oxidase. Infiltration of the fruit with aminooxyacetic acid, a potent inhibitor of the conversion of S-adenosylmethionine (AdoMet) to ACC completely blocked CO 2 stress-induced ethylene production. Similarly, cycloheximide, an inhibitor of nucleocytoplasmic protein synthesis effectively blocked CO 2 stress induction of polyamine accumulation, ethylene production, ACC formation and the development of ACC synthase. Withdrawal of CO 2 gas caused cessation of increases in ethylene production, respiration, ACC, putrescine and the activities of ACC synthase and ACC oxidase, but caused increase in spermidine and spermine levels. These data indicate that CO 2 induces de navo synthesis of ACC synthase thereby causing accumulation of ACC and increase in ethylene production and suggest that the conversion of AdoMet to ACC is the rate-limiting step in CO 2 stress-induced ethylene biosynthesis. The induction, however, requires continuous presence of the stimulus. The results also suggest that protein synthesis might be required for the CO 2 stress induction of polyamine biosynthesis. The results further suggest that in cucumber fruit under CO 2 stress, at least, the ethylene and polyamine biosynthetic pathways are not competitive.

Ethylene Biosynthesis and its Regulation in Higher Plants

PATHWAY OF ETHYLENE BIOSyNTHESIS 156 Methionine as an Intermediate ....... 157 S-Adenosylmethionine as an Intermediate 158 I-Aminocyclopropanecarboxylic Acid as an Intermediate . ......... 158 Methionine Cycle ........ 161 Conversion of l-Aminocyclopropanecarboxylic Acid to Ethylene 164 REGULATION OF ETHYLENE BIOSYNTHESIS 167 Regulation in Ripening Fruits and Senescing Flowers ...... 167 Auxin-Induced Ethylene Production 169 Regulation of Ethylene Biosynthesis by Ethylene 169 Stress-Induced Ethylene Production . . . . . . . . 171 Regulation by Light and Carbon Dioxide 173 Inhibitors of Ethylene Biosynthesis : 174 Conjugation of l-Aminocyclopropanecarboxylic Acid to l-(Malonylamino) cyclopropanecarboxylic Acid 178 CONCLUDING REMARKS 180