Ethylene Response Pathway Research Articles

‘Bartlett’ pear fruit (Pyrus communis L.) ripening regulation by low temperatures involves genes associated with jasmonic acid, cold response, and transcription factors

Low temperature (LT) treatments enhance ethylene production and ripening rate in the European pear (Pyrus communis L.). However, the underlying molecular mechanisms are not well understood. This study aims to identify genes responsible for ripening enhancement by LT. To this end, the transcriptome of ‘Bartlett’ pears treated with LT (0°C or 10°C for up to 14 d), which results in faster ripening, and control pears without conditioning treatment was analyzed. LT conditioned pears reached eating firmness (18N) in 6 d while control pears took about 12 d when left to ripen at 20°C. We identified 8,536 differentially expressed (DE) genes between the 0°C-treated and control fruit, and 7,938 DE genes between the 10°C-treated and control fruit. In an attempt to differentiate temperature-induced vs. ethylene-responsive pathways, we also monitored gene expression in fruit sequentially treated with 1-MCP then exposed to low temperature. This analysis revealed that genes associated with jasmonic acid biosynthesis and signaling, as well as the transcription factors TCP9a, TCP9b, CBF1, CBF4, AGL24, MYB1R1, and HsfB2b could be involved in the LT-mediated enhancement of ripening independently or upstream of ethylene.

Ethylene response pathway modulates attractiveness of plant roots to soybean cyst nematode Heterodera glycines

Plant parasitic nematodes respond to root exudates to locate their host roots. In our studies second stage juveniles of Heterodera glycines, the soybean cyst nematode (SCN), quickly migrated to soybean roots in Pluronic F-127 gel. Roots of soybean and non-host Arabidopsis treated with the ethylene (ET)-synthesis inhibitor aminoethoxyvinylglycine (AVG) were more attractive to SCN than untreated roots, and significantly more nematodes penetrated into roots. Moreover, Arabidopsis ET insensitive mutants (ein2, ein2-1, ein2-5, ein3-1, ein5-1, and ein6) were more attractive than wild-type plants. Conversely, the constitutive triple-response mutant ctr1-1, was less attractive to SCN. While ET receptor gain-of-function mutant ein4-1 attracted more SCN than the wild-type, there were no significant differences in attractiveness between another gain-of-function ET receptor mutant, etr1-3, or the loss-of-function mutants etr1-7 and ers1-3 and the wild type. Expression of the reporter construct EBS: β-glucuronidase (GUS) was detected in Arabidopsis root tips as early as 6 h post infection, indicating that ET signaling was activated in Arabidopsis early by SCN infection. These results suggest that an active ET signaling pathway reduces root attractiveness to SCN in a way similar to that reported for root-knot nematodes, but opposite to that suggested for the sugar beet cyst nematode Heterodera schachtii.

Transcriptome profiling analysis for two Tibetan wild barley genotypes in responses to low nitrogen.

BackgroundNitrogen (N) is the most common limiting factor for crop productivity worldwide. An effective approach to solve N deficiency is to develop low N (LN) tolerant crop cultivars. Tibetan annual wild barley is well-known for its wide genetic diversity and high tolerance to poor soil fertility. Up to date, no study has been done to illustrate the mechanism of LN tolerance underlying the wild barley at transcriptional level.ResultsIn this study, we employed Illumina RNA-Sequencing to determine the genotypic difference in transcriptome profile using two Tibetan wild barley genotypes differing in LN tolerance (XZ149, tolerant and XZ56, sensitive). A total of 1469 differentially expressed genes (DEGs) were identified in the two genotypes at 6 h and 48 h after LN treatment. Genetic difference existed in DEGs between XZ149 and XZ56, including transporters, transcription factors (TFs), kinases, antioxidant stress and hormone signaling related genes. Meanwhile, 695 LN tolerance-associated DEGs were mainly mapped to amino acid metabolism, starch and sucrose metabolism and secondary metabolism, and involved in transporter activity, antioxidant activities, and other gene ontology (GO). XZ149 had a higher capability of N absorption and use efficiency under LN stress than XZ56. The higher expression of nitrate transporters and energy-saving assimilation pattern could be attributed to its more N uptake and higher LN tolerance. In addition, auxin (IAA) and ethylene (ETH) response pathways may be also related to the genotypic difference in LN tolerance.ConclusionThe responses of XZ149 and XZ56 to LN stress differed dramatically at transcriptional level. The identified candidate genes related to LN tolerance may provide new insights into comprehensive understanding of the genotypic difference in N utilization and LN tolerance.Electronic supplementary materialThe online version of this article (doi:10.1186/s12870-016-0721-8) contains supplementary material, which is available to authorized users.

Ethylene Response Factors: A Key Regulatory Hub in Hormone and Stress Signaling.

Ethylene is essential for many developmental processes and a key mediator of biotic and abiotic stress responses in plants. The ethylene signaling and response pathway includes Ethylene Response Factors (ERFs), which belong to the transcription factor family APETALA2/ERF. It is well known that ERFs regulate molecular response to pathogen attack by binding to sequences containing AGCCGCC motifs (the GCC box), a cis-acting element. However, recent studies suggest that several ERFs also bind to dehydration-responsive elements and act as a key regulatory hub in plant responses to abiotic stresses. Here, we review some of the recent advances in our understanding of the ethylene signaling and response pathway, with emphasis on ERFs and their role in hormone cross talk and redox signaling under abiotic stresses. We conclude that ERFs act as a key regulatory hub, integrating ethylene, abscisic acid, jasmonate, and redox signaling in the plant response to a number of abiotic stresses.

Mitogen-Activated Protein Kinase 6 and Ethylene and Auxin Signaling Pathways Are Involved in Arabidopsis Root-System Architecture Alterations by Trichoderma atroviride.

Trichoderma atroviride is a symbiotic fungus that interacts with roots and stimulates plant growth and defense. Here, we show that Arabidopsis seedlings cocultivated with T. atroviride have an altered root architecture and greater biomass compared with axenically grown seedlings. These effects correlate with increased activity of mitogen-activated protein kinase 6 (MPK6). The primary roots of mpk6 mutants showed an enhanced growth inhibition by T. atroviride when compared with wild-type (WT) plants, while T. atroviride increases MPK6 activity in WT roots. It was also found that T. atroviride produces ethylene (ET), which increases with l-methionine supply to the fungal growth medium. Analysis of growth and development of WT seedlings and etr1, ein2, and ein3 ET-related Arabidopsis mutants indicates a role for ET in root responses to the fungus, since etr1 and ein2 mutants show defective root-hair induction and enhanced primary-root growth inhibition when cocultivated with T. atroviride. Increased MPK6 activity was evidenced in roots of ctr1 mutants, which correlated with repression of primary root growth, thus connecting MPK6 signaling with an ET response pathway. Auxin-inducible gene expression analysis using the DR5:uidA reporter construct further revealed that ET affects auxin signaling through the central regulator CTR1 and that fungal-derived compounds, such as indole-3-acetic acid and indole-3-acetaldehyde, induce MPK6 activity. Our results suggest that T. atroviride likely alters root-system architecture modulating MPK6 activity and ET and auxin action.

From root to fruit: RNA-Seq analysis shows that arbuscular mycorrhizal symbiosis may affect tomato fruit metabolism.

BackgroundTomato (Solanum lycopersicum) establishes a beneficial symbiosis with arbuscular mycorrhizal (AM) fungi. The formation of the mycorrhizal association in the roots leads to plant-wide modulation of gene expression. To understand the systemic effect of the fungal symbiosis on the tomato fruit, we used RNA-Seq to perform global transcriptome profiling on Moneymaker tomato fruits at the turning ripening stage.ResultsFruits were collected at 55 days after flowering, from plants colonized with Funneliformis mosseae and from control plants, which were fertilized to avoid responses related to nutrient deficiency. Transcriptome analysis identified 712 genes that are differentially expressed in fruits from mycorrhizal and control plants. Gene Ontology (GO) enrichment analysis of these genes showed 81 overrepresented functional GO classes. Up-regulated GO classes include photosynthesis, stress response, transport, amino acid synthesis and carbohydrate metabolism functions, suggesting a general impact of fungal symbiosis on primary metabolisms and, particularly, on mineral nutrition. Down-regulated GO classes include cell wall, metabolism and ethylene response pathways. Quantitative RT-PCR validated the RNA-Seq results for 12 genes out of 14 when tested at three fruit ripening stages, mature green, breaker and turning. Quantification of fruit nutraceutical and mineral contents produced values consistent with the expression changes observed by RNA-Seq analysis.ConclusionsThis RNA-Seq profiling produced a novel data set that explores the intersection of mycorrhization and fruit development. We found that the fruits of mycorrhizal plants show two transcriptomic “signatures”: genes characteristic of a climacteric fleshy fruit, and genes characteristic of mycorrhizal status, like phosphate and sulphate transporters. Moreover, mycorrhizal plants under low nutrient conditions produce fruits with a nutrient content similar to those from non-mycorrhizal plants under high nutrient conditions, indicating that AM fungi can help replace exogenous fertilizer for fruit crops.Electronic supplementary materialThe online version of this article (doi:10.1186/1471-2164-15-221) contains supplementary material, which is available to authorized users.

A new genetic linkage map of barley (Hordeum vulgare L.) facilitates genetic dissection of height and spike length and angle

Plant height and spike length and angle are important agronomic traits in the production of barley (Hordeum vulgare L.) due to strong correlations with lodging and disease. The objective of this study was to use QTL analysis to identify genetic regions associated with each trait in a recombinant inbred line (RIL) mapping population derived from a cross of Falcon by Azhul. Falcon is a spring six-row hulless feed barley with long spikes displaying obtuse angles, while Azhul is a spring dwarf, six-row hulless food barley with short spikes displaying acute angles. The population was genotyped using SNP, DArT and SSR markers and quantitative trait loci (QTL) were detected on chromosomes 2H (102.8cM, spike length), 3H (89.2cM, plant height and 38.2, spike angle and length), 4H (19.0cM, spike length), and 5H (106.7cM, spike angle). In conclusion, we developed a barley genetic map, which incorporated SNP, DArT and SSR markers, for detection of height and spike length and angle QTL. Three spike angle, one spike length and one plant height QTL were novel and by using comparative genomics we identified possible candidate genes involved in gibberellic acid signaling and auxin- and ethylene-responsive pathways. This knowledge can be used to generate suitable markers for barley breeding improvement.

A resistance mechanism dependent upon the inhibition of ethylene biosynthesis

The highly selective auxin-type herbicide quinclorac is widely used to control important dicotyledon and monocotyledon weeds in rice fields. Echinochloa crusgalli var. zelayensis is one of the most troublesome weeds in China, and is very difficult to control in east China due to misuse of herbicides. The JZD -R, JTJ -R, JCW -R and SSX -R biotypes of E. crusgalli var. zelayensis had resistance to quinclorac with resistance levels ranked as JZD -R < JTJ -R < JCW -R < SSX -R. Growth reduction in different biotypes was positively correlated with ethylene production. Stimulated levels of ethylene and 1-aminocyclopropane-1-carboxylic acid (ACC) and activities of ACC synthase and ACC oxidase in resistant biotypes were less than the susceptible biotype, and were negatively correlated with quinclorac resistance levels, suggesting that inhibition of ethylene biosynthesis was positively correlated with resistance levels. Considering the resistance-dependent inhibition in the ethylene biosynthetic pathway, the mechanisms of resistance to quinclorac in E. crusgalli var. zelayensis involved alteration(s) in the ethylene response pathway, consisting of at least alteration in induction of the enzymes activity of ACC synthase and ACC oxidase.

Identification of two-component system elements downstream of AHK5 in the stomatal closure response of Arabidopsis thaliana

To optimize water use efficiency, plants regulate stomatal closure through a complex signaling process. Hydrogen peroxide (H2O2) is produced in response to several environmental stimuli, and has been identified as a key second messenger involved in the regulation of stomatal aperture. The Arabidopsis histidine kinase 5 (AHK5) has been shown to regulate stomatal closure in response to H2O2 and other stimuli that depend on H2O2. AHK5 is a member of the two-component system (TCS) in Arabidopsis. The plant TCS comprises three different protein types: the hybrid histidine kinases (HKs), the phosphotransfer proteins (HPs) and the response regulators (RRs). Here we determined TCS elements involved in H2O2- and ethylene-dependent stomatal closure downstream of AHK5. By yeast and in planta interaction assays and functional studies, AHP1, 2 and 5 as well as the response regulators ARR4 and ARR7 were identified acting downstream of AHK5 in the ethylene and H2O2 response pathways of guard cells. Furthermore, we demonstrate that aspartate phosphorylation of ARR4 is only required for the H2O2- but not for the ethylene-induced stomatal closure response. Our data suggest the presence of a complex TCS signaling network comprising of at least AHK5, several AHPs and response regulators, which modulate stomatal closure in response to H2O2 and ethylene.

Pseudonodule Formation by Wild-Type and Symbiotic Mutant Medicago truncatula in Response to Auxin Transport Inhibitors

Rhizobium and allied bacteria form symbiotic nitrogen-fixing nodules on legume roots. Plant hormones play key roles in nodule formation. We treated Medicago truncatula roots with auxin transport inhibitors (ATI) N-(1-naphthyl)phthalamic acid (NPA) and 2,3,5-triiodobenzoic acid (TIBA) to induce the formation of pseudonodules. M. truncatula mutants defective for rhizobial Nod factor signal transduction still formed pseudonodules in response to ATI. However, a M. truncatula ethylene-insensitive supernodulator, sickle 1-1, did not form pseudonodules in response to TIBA, suggesting that the ethylene response pathway is involved in ATI-induced pseudonodule formation. We compared the transcriptional responses of M. truncatula roots treated with ATI to roots inoculated with Sinorhizobium meliloti. Some genes showed consistently parallel expression in ATI-induced and Rhizobium-induced nodules. For other genes, the transcriptional response of M. truncatula roots 1 and 7 days after ATI treatment was in the opposite direction to roots treated with S. meliloti; then, by 21 days, the transcriptional patterns for the two conditions became similar. We silenced 17 genes that were upregulated in both ATI and S. meliloti treatments to determine their effect on nodule formation. Some gene-silenced roots showed a decrease in nodulation efficiency, suggesting a role in nodule formation but not in later nodule functions.

Effect of seed on ripening control components during avocado fruit development

Seedless avocado fruit are produced alongside seeded fruit in the cultivar Arad, and both reach maturity at the same time. Using this system, it was possible to show that avocado seed inhibits the ripening process: seedless fruits exhibited higher response to exogenous ethylene already at the fruitlet stage, and also at the immature and mature fruit stages. They produced higher CO 2 levels, and the ethylene peak was apparent at the fruitlet stage of seedless fruit, but not of seeded ones. The expression levels of PaETR, PaERS1 and PaCTR1 on the day of harvest at all developmental stages were very similar between seeded and seedless fruit, except that PaCTR1 was higher in seedless fruit only at very early stages. This expression pattern suggests that the seed does not have an effect on components of the ethylene response pathway when fruits are just picked. The expression of MADS-box genes, PaAG1 and PaAGL9, preceded the increase in ethylene production of mature seeded fruit, but not at earlier stages. However, only PaAGL9 was induced in seedless fruit at early stages of development. Taken together, these data suggest that these genes are perhaps involved in climacteric response in seeded fruit, and the seed is responsible for their induction at normal fruit ripening.

Responses of root hair development to elevated CO2

This review highlights a potential signaling pathway of CO2-dependent stimulation in root hair development. Elevated CO2 firstly increases the carbohydrates production, which triggers the auxin or ethylene responsive signal transduction pathways and subsequently stimulates the generation of intracellular nitric oxide (NO). The NO acts on target Ca2+ and ion channels and induces activation of MAPK. Meanwhile, reactive oxygen species (ROS) activates cytoplasmic Ca2+ channels at the plasma membrane in the apex of the root tip. This complex pathway involves transduction cascades of multiple signals that lead to the fine tuning of epidermal cell initiation and elongation. The results suggest that elevated CO2 plays an important role in cell differentiation processes at the root epidermis.

XAP5 CIRCADIAN TIMEKEEPER Regulates Ethylene Responses in Aerial Tissues of Arabidopsis

The phytohormone ethylene differentially regulates plant architecture and growth in both a light- and nutrient-dependent fashion. The modulation of plant development by ethylene in response to both external and internal signals can also generate tissue-specific differential responses. Here, we report that XAP5 CIRCADIAN TIMEKEEPER (XCT) is involved in blue light-dependent ethylene responses in the aerial tissues of Arabidopsis (Arabidopsis thaliana) seedlings. XCT was first identified as a circadian clock mutant with a short free-running period. The xct mutation also causes sugar-specific hypocotyl growth defects, in which mutants are short in blue light when grown on a sucrose-rich medium but tall when grown on sucrose-deficient medium. Our data suggest that the hypocotyl defects in blue light are not directly caused by defects in clock or light signaling but rather by enhanced ethylene responses. In blue light, xct mutants have a more active ethylene response pathway and exhibit growth phenotypes similar to the constitutive ethylene signaling mutant constitutive triple response1 (ctr1). xct mutants also have reduced ethylene emission, analogous to plants that have lost CTR1 function. Genetic analysis suggests that XCT negatively regulates ethylene responses downstream of ETHYLENE-INSENSITIVE3 in aerial tissues. However, XCT is not required for all ethylene-mediated processes, such as the inhibition of root growth. Thus, XCT acts downstream of a major transcriptional regulator in an organ-specific manner, playing an environment-dependent role in the regulation of plant growth.

Characterization of opaque2 modifier QTLs and candidate genes in recombinant inbred lines derived from the K0326Y quality protein maize inbred

Quality protein maize (QPM) is a high lysine-containing corn that is based on genetic modification of the opaque2 (o2) mutant. In QPM, modifier genes convert the starchy endosperm of o2 to the vitreous phenotype of wild type maize. There are multiple, unlinked o2 modifier loci (Opm) in QPM and their nature and mode of action are unknown. We previously identified seven Opm QTLs and characterized 16 genes that are differentially up-regulated at a significant level in K0326Y QPM, compared to the starchy endosperm mutant W64Ao2. In order to further characterize these Opm QTLs and the genes up-regulated in K0326Y QPM, we created a population of 314 recombinant inbred lines (RILs) from a cross between K0326Y QPM and W64Ao2. The RILs were characterized for three traits associated with endosperm texture: vitreousness, density and hardness. Genetic linkage analysis of the RIL population confirmed three of the previously identified QTLs associated with o2 endosperm modification in K0326Y QPM. Many of the genes up-regulated in K0326Y QPM showed substantially higher levels of expression in vitreous compared with opaque RILs. These included genes associated with the upstream regulation of the ethylene response pathway, and a gene encoding a regulatory subunit of pyrophosphate-dependent fructose-6-phosphate 1-phosphotransferase, an adaptive enzyme of the glycolytic pathway.

The Copper Transporter RAN1 Is Essential for Biogenesis of Ethylene Receptors in Arabidopsis

Plants utilize ethylene as a hormone to regulate multiple developmental processes and to coordinate responses to biotic and abiotic stress. In Arabidopsis thaliana, a small family of five receptor proteins typified by ETR1 mediates ethylene perception. Our previous work suggested that copper ions likely play a role in ethylene binding. An independent study indicated that the ran1 mutants, which display ethylene-like responses to the ethylene antagonist trans-cyclooctene, have mutations in the RAN1 copper-transporting P-type ATPase, once again linking copper ions to the ethylene-response pathway. The results presented herein indicate that genetically engineered Saccharomyces cerevisiae expressing ETR1 but lacking the RAN1 homolog Ccc2p (Δccc2) lacks ethylene-binding activity. Ethylene-binding activity was restored when copper ions were added to the Δccc2 mutants, showing that it is the delivery of copper that is important. Additionally, transformation of the Δccc2 mutant yeast with RAN1 rescued ethylene-binding activity. Analysis of plants carrying loss-of-function mutations in ran1 showed that they lacked ethylene-binding activity, whereas seedlings carrying weak alleles of ran1 had normal ethylene-binding activity but were hypersensitive to copper-chelating agents. Altogether, the results show an essential role for RAN1 in the biogenesis of the ethylene receptors and copper homeostasis in Arabidopsis seedlings. Furthermore, the results indicate cross-talk between the ethylene-response pathway and copper homeostasis in Arabidopsis seedling development.

Abscisic acid-induced resistance against the brown spot pathogen Cochliobolus miyabeanus in rice involves MAP kinase-mediated repression of ethylene signaling.

The plant hormone abscisic acid (ABA) is involved in an array of plant processes, including the regulation of gene expression during adaptive responses to various environmental cues. Apart from its well-established role in abiotic stress adaptation, emerging evidence indicates that ABA is also prominently involved in the regulation and integration of pathogen defense responses. Here, we demonstrate that exogenously administered ABA enhances basal resistance of rice (Oryza sativa) against the brown spot-causing ascomycete Cochliobolus miyabeanus. Microscopic analysis of early infection events in control and ABA-treated plants revealed that this ABA-inducible resistance (ABA-IR) is based on restriction of fungal progression in the mesophyll. We also show that ABA-IR does not rely on boosted expression of salicylic acid-, jasmonic acid -, or callose-dependent resistance mechanisms but, instead, requires a functional Galpha-protein. In addition, several lines of evidence are presented suggesting that ABA steers its positive effect on brown spot resistance through antagonistic cross talk with the ethylene (ET) response pathway. Exogenous ethephon application enhances susceptibility, whereas genetic disruption of ET signaling renders plants less vulnerable to C. miyabeanus attack, thereby inducing a level of resistance similar to that observed on ABA-treated wild-type plants. Moreover, ABA treatment alleviates C. miyabeanus-induced activation of the ET reporter gene EBP89, while derepression of pathogen-triggered EBP89 transcription via RNA interference-mediated knockdown of OsMPK5, an ABA-primed mitogen-activated protein kinase gene, compromises ABA-IR. Collectively, these data favor a model whereby exogenous ABA enhances resistance against C. miyabeanus at least in part by suppressing pathogen-induced ET action in an OsMPK5-dependent manner.

Ethylene regulation of avocado ripening differs between seeded and seedless fruit

We studied the contribution of the seed to avocado ripening, emphasizing its role in ethylene biosynthesis and response pathways. Transcription profiles of genes involved in ethylene biosynthesis ( PaACO, PaACS1 and PaACS2) and action ( PaETR, PaERS1 and PaCTR1) were studied in seeded and seedless avocado fruit during ripening at ambient and low temperatures and in response to exogenous ethylene and 1-methylcyclopropene (1-MCP). Seedless mature fruit had a shorter preclimacteric lag, faster softening, and higher respiration during ambient temperature ripening than seeded ones. Advanced ripening in seedless fruit was accompanied by higher levels of PaACO and PaACS1 expression at harvest, and these levels increased dramatically towards the climacteric peak. The expression of PaETR, PaERS1 and PaCTR1 increased in parallel with the onset of the ethylene burst in seedless fruit, whereas PaETR increase predominantly in seeded ones. Seedless fruit exhibited an earlier response to exogenous ethylene at the day of harvest, than seeded fruit. On day 1 after harvest, ethylene application elicited lower levels of ethylene in seeded than in seedless fruit, concomitantly with massive PaCTR1 augmentation. This suggests that the negative regulator PaCTR may moderate the effect of ethylene on seeded fruit. Cold storage induced biosynthesis and regulatory genes in both seedless and seeded fruit relative to their levels at ambient temperature. However, in the first and second weeks in cold storage, PaACO, PaACS1 and PaACS2 expression levels were much higher in seedless than in seeded fruit, which could explain the higher levels of ethylene and accelerated softening of the seedless fruit in cold storage. Seeded and seedless fruit responded similarly to ethylene or 1-MCP application prior to cold storage. Ethylene slightly induced ethylene production, but significantly increased CO 2 output. 1-MCP equally and effectively delayed softening, reduced ethylene and CO 2 production and expression of genes involved in ethylene biosynthesis and ethylene action in seeded and seedless fruit. Both at ambient temperature and in cold storage respiration was higher in seedless than seeded fruit. Our findings demonstrate that the seed is involved in regulation of ethylene responsiveness during ripening, and acts to delay climacteric in mature seeded fruit.

Identification of Arabidopsis stay-green mutants with a functional ethylene-response pathway

Natural or harvest-induced senescence is a major determinant factor causing crop losses. The plant hormone ethylene is a strong inducer of senescence and decreasing the ethylene response can reduce senescence, albeit often with undesirable pleiotropic effects. We took advantage of ethylene-induced leaf senescence as a tool to screen for late senescence Arabidopsis mutants that still have a functional ethylenesignalling pathway. Sixteen Arabidopsis onset of leaf death (old) mutants were selected that stayed green after treatment with ethylene. While all the mutants responded to ethylene in a triple response assay, ten mutants responded to the treatment in the same way as the wild type. These ten mutants showed limited pleiotropic effects when grown under standard growth conditions but nine mutants flowered slightly later than the wild type. Genetic characterisation of a subset of the mutants identified several independent loci controlling the leaf senescence process. The approach resulted in the isolation of several stay-green mutants with a functional ethylene response pathway. The late senescence mutants show extended leaf longevity and further research may advance the field of pre- or post-harvest senescence technology. The results, moreover, suggest that there is a correlation between senescence and floral induction. Keywords: Senescence, Arabidopsis, ethylene, mutant, shelf life

1-Aminocyclopropane-1-carboxylic acid (ACC) concentration and ACC synthase expression in soybean roots, root tips, and soybean cyst nematode (Heterodera glycines)-infected roots

Colonization of plant roots by root knot and cyst nematodes requires a functional ethylene response pathway. However, ethylene plays many roles in root development and whether its role in nematode colonization is direct or indirect, for example lateral root initiation or root hair growth, is not known. The temporal requirement for ethylene and localized synthesis of ethylene during the life span of soybean cyst nematode (SCN) on soybean roots was further investigated. Although a significant increase in ethylene evolution was not detected from SCN-colonized roots, the concentration of the immediate precursor to ethylene, 1-aminocyclopropane-1-carboxylic acid (ACC), was higher in SCN-colonized root pieces and root tips than in other parts of the root. Moreover, expression analysis of 17 ACC synthase (ACS) genes indicated that a select set of ACS genes is expressed in SCN-colonized root pieces that is clearly different from the set of genes expressed in non-colonized roots or root tips. Semi-quantitative real-time PCR indicated that ACS transcript accumulation correlates with the high concentration of ACC in root tips. In addition, an ACS-like sequence was found in the public SCN nucleotide database. Acquisition of a full-length sequence for this mRNA (accession GQ389647) and alignment with transcripts for other well-characterized ACS proteins indicated that the nematode sequence is missing a key element required for ACS activity and therefore probably is not a functional ACS. Moreover, no significant amount of ACC was found in any growth stage of SCN that was tested.

Recent advances in ethylene research

Ethylene regulates many aspects of the plant life cycle, including seed germination, root initiation, flower development, fruit ripening, senescence, and responses to biotic and abiotic stresses. It thus plays a key role in responses to the environment that have a direct bearing on a plant's fitness for adaptation and reproduction. In recent years, there have been major advances in our understanding of the molecular mechanisms regulating ethylene synthesis and action. Screening for mutants of the triple response phenotype of etiolated Arabidopsis seedlings, together with map-based cloning and candidate gene characterization of natural mutants from other plant species, has led to the identification of many new genes for ethylene biosynthesis, signal transduction, and response pathways. The simple chemical nature of ethylene contrasts with its regulatory complexity. This is illustrated by the multiplicity of genes encoding the key ethylene biosynthesis enzymes 1-aminocyclopropane-1-carboxylic acid (ACC) synthase and ACC oxidase, multiple ethylene receptors and signal transduction components, and the complexity of regulatory steps involving signalling relays and control of mRNA and protein synthesis and turnover. In addition, there are extensive interactions with other hormones. This review integrates knowledge from the model plant Arabidopsis and other plant species and focuses on key aspects of recent research on regulatory networks controlling ethylene synthesis and its role in flower development and fruit ripening.