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1.
J Exp Bot ; 73(18): 6115-6132, 2022 10 18.
Artículo en Inglés | MEDLINE | ID: mdl-35639812

RESUMEN

Small secreted peptides have been described as key contributors to complex signalling networks that control plant development and stress responses. The Brassicaceae-specific PROSCOOP family encodes precursors of Serine riCh endOgenOus Peptides (SCOOPs). In Arabidopsis SCOOP12 has been shown to promote the defence response against pathogens and to be involved in root development. Here, we explore its role as a moderator of Arabidopsis primary root development. We show that the PROSCOOP12 null mutation leads to longer primary roots through the development of longer differentiated cells while PROSCOOP12 overexpression induces dramatic plant growth impairments. In comparison, the exogenous application of synthetic SCOOP12 peptide shortens roots through meristem size and cell length reductions. Moreover, superoxide anion (O2·-) and hydrogen peroxide (H2O2) production in root tips vary according to SCOOP12 abundance. By using reactive oxygen species scavengers that suppress the proscoop12 phenotype, we showed that root growth regulation by SCOOP12 is associated with reactive oxygen species metabolism. Furthermore, our results suggest that peroxidases act as potential SCOOP12 downstream targets to regulate H2O2 production, which in turn triggers cell wall modifications in root. Finally, a massive transcriptional reprogramming, including the induction of genes from numerous other pathways, including ethylene, salicylic acid, and glucosinolates biosynthesis, was observed, emphasizing its dual role in defence and development.


Asunto(s)
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Peróxido de Hidrógeno/metabolismo , Superóxidos/metabolismo , Glucosinolatos/metabolismo , Raíces de Plantas/metabolismo , Regulación de la Expresión Génica de las Plantas , Etilenos/metabolismo , División Celular , Homeostasis , Péptidos/metabolismo , Ácido Salicílico/metabolismo , Peroxidasas/genética , Serina/metabolismo
2.
Mol Genet Genomics ; 295(6): 1443-1457, 2020 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-32700103

RESUMEN

The apple is a highly perishable fruit after harvesting and, therefore, several storage technologies have been studied to provide the consumer market with a quality product with a longer shelf life. However, little is known about the apple genome that is submitted to the storage, and even less with the application of ripening inhibitors. Due to these factors, this study sought to elucidate the transcriptional profile of apple cultivate Gala stored in a controlled atmosphere (AC) treated and not treated with 1-methyl cyclopropene (1-MCP). Through the genetic mapping of the apple, applying the microarray technique, it was possible to verify the action of treatments on transcripts related to photosynthesis, carbohydrate metabolism, response to hormonal stimuli, nucleic acid metabolism, reduction of oxidation, regulation of transcription and metabolism of cell wall and lipids. The results showed that the transcriptional profile in the entire genome of the fruit showed significant differences in the relative expression of the gene, this in response to CA in the presence and absence of 1-MCP. It should be noted that the transcription genes involved in the anabolic pathway were only maintained after six months in fruits treated with 1-MCP. The data in this work suggests that the apple in the absence of 1-MCP begins to prepare its metabolism to mature, even during the storage period in AC. Meanwhile, in the presence of the inhibitor, the transcriptional profile of the fruit is similar to that at the time of harvest. It was also found that a set of genes that code for ethylene receptors, auxin homeostasis, MADS Box, and NAC transcription factors may be involved in the regulation of post-harvest ripening after storage and in the absence of 1-MCP.


Asunto(s)
Ciclopropanos/metabolismo , Frutas/metabolismo , Regulación de la Expresión Génica de las Plantas , Malus/metabolismo , Proteínas de Plantas/metabolismo , Factores de Transcripción/metabolismo , Almacenamiento de Alimentos , Frutas/crecimiento & desarrollo , Malus/crecimiento & desarrollo , Proteínas de Plantas/genética , Factores de Transcripción/genética
3.
Mol Cell ; 48(1): 109-20, 2012 Oct 12.
Artículo en Inglés | MEDLINE | ID: mdl-22940249

RESUMEN

In Arabidopsis thaliana, the putative RNA-helicase SDE3 assists posttranscriptional-gene-silencing (PTGS) amplification by RNA-dependent-RNA-polymerase-6 (RDR6). SDE3 homologs in Drosophila, worm and human contribute to silence viruses, transposons or recently duplicated genes but the underlying mechanisms remain largely unknown. Here, we demonstrate that SDE3 is present with the PTGS effectors AGO1 and AGO2 in higher-order protein complexes owing to a specialized GW-repeat-containing C-terminal domain. We uncover an essential contribution of the RNA-helicase activity and a facilitating role for AGO binding in SDE3 action, which occurs downstream of RDR6. We show that these biochemical properties underpin dual roles for SDE3 in antiviral defense and, unexpectedly, in transposon silencing via a hitherto unanticipated pathway that correlates with DNA methylation, suggesting a continuum of action between PTGS and chromatin-level silencing. We identified endogenous SDE3 targets corresponding to nonconserved intergenic regions, transposons and recently evolved pseudogenes, unraveling striking functional convergences among plant and metazoan SDE3 pathways.


Asunto(s)
Proteínas de Arabidopsis/química , Proteínas de Arabidopsis/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , ADN Intergénico/genética , ADN de Plantas/genética , ARN Helicasas/química , ARN Helicasas/metabolismo , Secuencias de Aminoácidos , Secuencia de Aminoácidos , Animales , Arabidopsis/virología , Proteínas de Arabidopsis/genética , Proteínas Argonautas/química , Proteínas Argonautas/metabolismo , Secuencia Conservada , Metilación de ADN , ADN Intergénico/metabolismo , ADN de Plantas/metabolismo , Humanos , Modelos Biológicos , Datos de Secuencia Molecular , Dominios y Motivos de Interacción de Proteínas , ARN Helicasas/genética , Interferencia de ARN , Proteínas de Unión al ARN/química , Proteínas de Unión al ARN/metabolismo , ARN Polimerasa Dependiente del ARN/metabolismo , Homología de Secuencia de Aminoácido
4.
J Exp Bot ; 70(4): 1349-1365, 2019 02 20.
Artículo en Inglés | MEDLINE | ID: mdl-30715439

RESUMEN

Small secreted peptides are important players in plant development and stress response. Using a targeted in silico approach, we identified a family of 14 Arabidopsis genes encoding precursors of serine-rich endogenous peptides (PROSCOOP). Transcriptomic analyses revealed that one member of this family, PROSCOOP12, is involved in processes linked to biotic and oxidative stress as well as root growth. Plants defective in this gene were less susceptible to Erwinia amylovora infection and showed an enhanced root growth phenotype. In PROSCOOP12 we identified a conserved motif potentially coding for a small secreted peptide. Exogenous application of synthetic SCOOP12 peptide induces various defense responses in Arabidopsis. Our findings show that SCOOP12 has numerous properties of phytocytokines, activates the phospholipid signaling pathway, regulates reactive oxygen species response, and is perceived in a BAK1 co-receptor-dependent manner.


Asunto(s)
Proteínas de Arabidopsis/fisiología , Arabidopsis/genética , Regulación de la Expresión Génica de las Plantas/inmunología , Genes de Plantas , Péptidos y Proteínas de Señalización Intercelular/fisiología , Familia de Multigenes , Raíces de Plantas/crecimiento & desarrollo , Arabidopsis/crecimiento & desarrollo , Arabidopsis/inmunología , Proteínas de Arabidopsis/genética , Péptidos y Proteínas de Señalización Intercelular/genética , Raíces de Plantas/genética , Transducción de Señal
6.
Nucleic Acids Res ; 43(Database issue): D1010-7, 2015 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-25392409

RESUMEN

CATdb (http://urgv.evry.inra.fr/CATdb) is a database providing a public access to a large collection of transcriptomic data, mainly for Arabidopsis but also for other plants. This resource has the rare advantage to contain several thousands of microarray experiments obtained with the same technical protocol and analyzed by the same statistical pipelines. In this paper, we present GEM2Net, a new module of CATdb that takes advantage of this homogeneous dataset to mine co-expression units and decipher Arabidopsis gene functions. GEM2Net explores 387 stress conditions organized into 18 biotic and abiotic stress categories. For each one, a model-based clustering is applied on expression differences to identify clusters of co-expressed genes. To characterize functions associated with these clusters, various resources are analyzed and integrated: Gene Ontology, subcellular localization of proteins, Hormone Families, Transcription Factor Families and a refined stress-related gene list associated to publications. Exploiting protein-protein interactions and transcription factors-targets interactions enables to display gene networks. GEM2Net presents the analysis of the 18 stress categories, in which 17,264 genes are involved and organized within 681 co-expression clusters. The meta-data analyses were stored and organized to compose a dynamic Web resource.


Asunto(s)
Arabidopsis/genética , Bases de Datos Genéticas , Regulación de la Expresión Génica de las Plantas , Redes Reguladoras de Genes , Estrés Fisiológico/genética , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Perfilación de la Expresión Génica , Internet , Modelos Genéticos , Mapeo de Interacción de Proteínas
7.
Funct Integr Genomics ; 16(2): 183-201, 2016 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-26797431

RESUMEN

In many plant/pathogen interactions, host susceptibility factors are key determinants of disease development promoting pathogen growth and spreading in plant tissues. In the Fusarium head blight (FHB) disease, the molecular basis of wheat susceptibility is still poorly understood while it could provide new insights into the understanding of the wheat/Fusarium graminearum (Fg) interaction and guide future breeding programs to produce cultivars with sustainable resistance. To identify the wheat grain candidate genes, a genome-wide gene expression profiling was performed in the French susceptible wheat cultivar, Recital. Gene-specific two-way ANOVA of about 40 K transcripts at five grain developmental stages identified 1309 differentially expressed genes. Out of these, 536 were impacted by the Fg effect alone. Most of these Fg-responsive genes belonged to biological and molecular functions related to biotic and abiotic stresses indicating the activation of common stress pathways during susceptibility response of wheat grain to FHB. This analysis revealed also 773 other genes displaying either specific Fg-responsive profiles along with grain development stages or synergistic adjustments with the grain development effect. These genes were involved in various molecular pathways including primary metabolism, cell death, and gene expression reprogramming. An increasingly complex host response was revealed, as was the impact of both Fg infection and grain ontogeny on the transcription of wheat genes. This analysis provides a wealth of candidate genes and pathways involved in susceptibility responses to FHB and depicts new clues to the understanding of the susceptibility determinism in plant/pathogen interactions.


Asunto(s)
Grano Comestible/genética , Fusarium/crecimiento & desarrollo , Regulación de la Expresión Génica de las Plantas , Enfermedades de las Plantas/genética , Transcriptoma , Triticum/genética , Resistencia a la Enfermedad , Susceptibilidad a Enfermedades , Grano Comestible/crecimiento & desarrollo , Grano Comestible/inmunología , Grano Comestible/microbiología , Fusarium/patogenicidad , Perfilación de la Expresión Génica , Ontología de Genes , Genoma de Planta , Interacciones Huésped-Patógeno , Redes y Vías Metabólicas/genética , Anotación de Secuencia Molecular , Enfermedades de las Plantas/inmunología , Enfermedades de las Plantas/microbiología , Proteínas de Plantas/genética , Triticum/crecimiento & desarrollo , Triticum/inmunología , Triticum/microbiología
8.
BMC Plant Biol ; 16(1): 201, 2016 09 15.
Artículo en Inglés | MEDLINE | ID: mdl-27630120

RESUMEN

BACKGROUND: Fruit quality depends on a series of biochemical events that modify appearance, flavour and texture throughout fruit development and ripening. Cell wall polysaccharide remodelling largely contributes to the elaboration of fleshy fruit texture. Although several genes and enzymes involved in cell wall polysaccharide biosynthesis and modifications are known, their coordinated activity in these processes is yet to be discovered. RESULTS: Combined transcriptomic and biochemical analyses allowed the identification of putative enzymes and related annotated members of gene families involved in cell wall polysaccharide composition and structural changes during apple fruit growth and ripening. The early development genes were mainly related to cell wall biosynthesis and degradation with a particular target on hemicelluloses. Fine structural evolutions of galactoglucomannan were strongly correlated with mannan synthase, glucanase (GH9) and ß-galactosidase gene expression. In contrast, fewer genes related to pectin metabolism and cell expansion (expansin genes) were observed in ripening fruit combined with expected changes in cell wall polysaccharide composition. CONCLUSIONS: Hemicelluloses undergo major structural changes particularly during early fruit development. The high number of early expressed ß-galactosidase genes questions their function on galactosylated structures during fruit development and storage. Their activity and cell wall substrate remains to be identified. Moreover, new insights into the potential role of peroxidases and transporters, along with cell wall metabolism open the way to further studies on concomitant mechanisms involved in cell wall assembly/disassembly during fruit development and storage.


Asunto(s)
Pared Celular/metabolismo , Frutas/crecimiento & desarrollo , Regulación de la Expresión Génica de las Plantas , Malus/genética , Polisacáridos/metabolismo , Pared Celular/genética , Frutas/genética , Frutas/metabolismo , Regulación del Desarrollo de la Expresión Génica , Malus/crecimiento & desarrollo , Malus/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo
9.
Plant Cell ; 25(7): 2444-63, 2013 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-23903317

RESUMEN

The molecular mechanisms underlying plant cell totipotency are largely unknown. Here, we present a protocol for the efficient regeneration of plants from Arabidopsis thaliana protoplasts. The specific liquid medium used in our study leads to a high rate of reentry into the cell cycle of most cell types, providing a powerful system to study dedifferentiation/regeneration processes in independent somatic cells. To identify the early events in the establishment of totipotency, we monitored the genome-wide transcript profiles of plantlets and protoplast-derived cells (PdCs) during the first week of culture. Plant cells rapidly dedifferentiated. Then, we observed the reinitiation and reorientation of protein synthesis, accompanied by the reinitiation of cell division and de novo cell wall synthesis. Marked changes in the expression of chromatin-associated genes, especially of those in the histone variant family, were observed during protoplast culture. Surprisingly, the epigenetic status of PdCs and well-established cell cultures differed, with PdCs exhibiting rare reactivated transposons and epigenetic changes. The differentially expressed genes identified in this study are interesting candidates for investigating the molecular mechanisms underlying plant cell plasticity and totipotency. One of these genes, the plant-specific transcription factor ABERRANT LATERAL ROOT FORMATION4, is required for the initiation of protoplast division.


Asunto(s)
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Protoplastos/metabolismo , Células Madre Totipotentes/metabolismo , Factores de Transcripción/genética , Transcriptoma/genética , Arabidopsis/citología , Proteínas de Arabidopsis/metabolismo , Proteínas de Arabidopsis/fisiología , Ciclo Celular/genética , Desdiferenciación Celular/genética , División Celular/genética , Células Cultivadas , Análisis por Conglomerados , Regulación del Desarrollo de la Expresión Génica , Ontología de Genes , Análisis de Secuencia por Matrices de Oligonucleótidos , Protoplastos/citología , Regeneración/genética , Regeneración/fisiología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factores de Tiempo , Células Madre Totipotentes/citología , Factores de Transcripción/metabolismo , Factores de Transcripción/fisiología
10.
Plant J ; 79(2): 322-33, 2014 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-24891222

RESUMEN

Faced with declining soil-water potential, plants synthesize abscisic acid (ABA), which then triggers stomatal closure to conserve tissue moisture. Closed stomates, however, also create several physiological dilemmas. Among these, the large CO2 influx required for net photosynthesis will be disrupted. Depleting CO2 in the plant will in turn bias stomatal opening by suppressing ABA sensitivity, which then aggravates transpiration further. We have investigated the molecular basis of how C3 plants resolve this H2 O-CO2 conflicting priority created by stomatal closure. Here, we have identified in Arabidopsis thaliana an early drought-induced spermidine spermine-N(1) -acetyltransferase homolog, which can slow ABA-mediated stomatal closure. Evidence from genetic, biochemical and physiological analyses has revealed that this protein does so by acetylating the metabolite 1,3-diaminopropane (DAP), thereby turning on the latter's intrinsic activity. Acetylated DAP triggers plasma membrane electrical and ion transport properties in an opposite way to those by ABA. Thus in adapting to low soil-water availability, acetyl-DAP could refrain stomates from complete closure to sustain CO2 diffusion to photosynthetic tissues.


Asunto(s)
Ácido Abscísico/metabolismo , Arabidopsis/metabolismo , Diaminas/metabolismo , Sequías , Estomas de Plantas/metabolismo , Proteínas de Arabidopsis/metabolismo , Regulación de la Expresión Génica de las Plantas , Transducción de Señal
11.
J Neurochem ; 132(6): 703-12, 2015 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-25533782

RESUMEN

Deep brain stimulation (DBS) of the subthalamic nucleus (STN) is an efficient neurosurgical treatment for advanced Parkinson's disease. Non-invasive metabolic neuroimaging during the course of DBS in animal models may contribute to our understanding of its action mechanisms. Here, DBS was adapted to in vivo proton magnetic resonance spectroscopy at 11.7 T in the rat to follow metabolic changes in main basal ganglia structures, the striatum, and the substantia nigra pars reticulata (SNr). Measurements were repeated OFF and ON acute and subchronic (7 days) STN-DBS in control and parkinsonian (6-hydroxydopamine lesion) conditions. Acute DBS reversed the increases in glutamate, glutamine, and GABA levels induced by the dopamine lesion in the striatum but not in the SNr. Subchronic DBS normalized GABA in both the striatum and SNr, and glutamate in the striatum. Taurine levels were markedly decreased under subchronic DBS in the striatum and SNr in both lesioned and unlesioned rats. Microdialysis in the striatum further showed that extracellular taurine was increased. These data reveal that STN-DBS has duration-dependent metabolic effects in the basal ganglia, consistent with development of adaptive mechanisms. In addition to counteracting defects induced by the dopamine lesion, prolonged DBS has proper effects independent of the pathological condition. Non-invasive metabolic neuroimaging might be useful to understand the physiological mechanisms of deep brain stimulation (DBS). Here, we demonstrate the feasibility of repeated high-field proton magnetic resonance spectroscopy of basal ganglia structures under subthalamic nucleus DBS in control and parkinsonian rats. Results show that DBS has both rapid and delayed effects either dependent or independent of disease state.


Asunto(s)
Ganglios Basales/metabolismo , Estimulación Encefálica Profunda/tendencias , Trastornos Parkinsonianos/metabolismo , Trastornos Parkinsonianos/terapia , Núcleo Subtalámico/metabolismo , Núcleo Subtalámico/cirugía , Animales , Encéfalo/metabolismo , Espectroscopía de Resonancia Magnética/métodos , Masculino , Ratas , Ratas Wistar , Factores de Tiempo
12.
Plant Physiol ; 164(4): 1930-51, 2014 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-24550240

RESUMEN

Apple (Malus × domestica) trees naturally produce an excess of fruitlets that negatively affect the commercial value of fruits brought to maturity and impact their capacity to develop flower buds the following season. Therefore, chemical thinning has become an important cultural practice, allowing the selective removal of unwanted fruitlets. As the public pressure to limit the use of chemical agents increases, the control of thinning becomes a major issue. Here, we characterized the self-thinning capacity of an apple hybrid genotype from the tree scale to the molecular level. Additional amphivasal vascular bundles were identified in the pith of pedicels supporting the fruitlets with the lowest abscission potential (central fruitlet), indicating that these bundles might have a role in the acquisition of dominance over lateral fruitlets. Sugar content analysis revealed that central fruitlets were better supplied in sorbitol than lateral fruitlets. Transcriptomic profiles allowed us to identify genes potentially involved in the overproduction of vascular tissues in central pedicels. In addition, histological and transcriptomic data permitted a detailed characterization of abscission zone development and the identification of key genes involved in this process. Our data confirm the major role of ethylene, auxin, and cell wall-remodeling enzymes in abscission zone formation. The shedding process in this hybrid appears to be triggered by a naturally exacerbated dominance of central fruitlets over lateral ones, brought about by an increased supply of sugars, possibly through additional amphivasal vascular bundles. The characterization of this genotype opens new perspectives for the selection of elite apple cultivars.


Asunto(s)
Frutas/anatomía & histología , Frutas/fisiología , Malus/anatomía & histología , Malus/fisiología , Haz Vascular de Plantas/fisiología , Metabolismo de los Hidratos de Carbono/genética , Análisis por Conglomerados , Frutas/genética , Perfilación de la Expresión Génica , Regulación del Desarrollo de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Genes de Plantas , Hibridación Genética , Cinética , Malus/genética , Modelos Biológicos , Fenotipo , Polinización/fisiología , Árboles/anatomía & histología , Árboles/genética , Árboles/fisiología
13.
Plant Physiol ; 164(4): 2167-83, 2014 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-24501001

RESUMEN

Siderophores are specific ferric iron chelators synthesized by virtually all microorganisms in response to iron deficiency. We have previously shown that they promote infection by the phytopathogenic enterobacteria Dickeya dadantii and Erwinia amylovora. Siderophores also have the ability to activate plant immunity. We have used complete Arabidopsis transcriptome microarrays to investigate the global transcriptional modifications in roots and leaves of Arabidopsis (Arabidopsis thaliana) plants after leaf treatment with the siderophore deferrioxamine (DFO). Physiological relevance of these transcriptional modifications was validated experimentally. Immunity and heavy-metal homeostasis were the major processes affected by DFO. These two physiological responses could be activated by a synthetic iron chelator ethylenediamine-di(o-hydroxyphenylacetic) acid, indicating that siderophores eliciting activities rely on their strong iron-chelating capacity. DFO was able to protect Arabidopsis against the pathogenic bacterium Pseudomonas syringae pv tomato DC3000. Siderophore treatment caused local modifications of iron distribution in leaf cells visible by ferrocyanide and diaminobenzidine-H2O2 staining. Metal quantifications showed that DFO causes a transient iron and zinc uptake at the root level, which is presumably mediated by the metal transporter iron regulated transporter1 (IRT1). Defense gene expression and callose deposition in response to DFO were compromised in an irt1 mutant. Consistently, plant susceptibility to D. dadantii was increased in the irt1 mutant. Our work shows that iron scavenging is a unique mechanism of immunity activation in plants. It highlights the strong relationship between heavy-metal homeostasis and immunity.


Asunto(s)
Arabidopsis/efectos de los fármacos , Arabidopsis/inmunología , Deferoxamina/farmacología , Hierro/metabolismo , Inmunidad de la Planta/efectos de los fármacos , Sideróforos/farmacología , Arabidopsis/genética , Arabidopsis/microbiología , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Proteínas de Transporte de Catión/genética , Proteínas de Transporte de Catión/metabolismo , Enterobacteriaceae/fisiología , Perfilación de la Expresión Génica , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Genes de Plantas , Homeostasis/efectos de los fármacos , Homeostasis/genética , Inmunidad Innata/efectos de los fármacos , Quelantes del Hierro/farmacología , Modelos Biológicos , Fosforilación/efectos de los fármacos , Enfermedades de las Plantas/microbiología , Hojas de la Planta/efectos de los fármacos , Hojas de la Planta/genética , Raíces de Plantas/efectos de los fármacos , Raíces de Plantas/genética , Pseudomonas syringae/efectos de los fármacos , Pseudomonas syringae/fisiología , Factores de Tiempo , Regulación hacia Arriba/efectos de los fármacos , Regulación hacia Arriba/genética , Agua/farmacología , Zinc/metabolismo
14.
J Exp Bot ; 66(9): 2659-72, 2015 May.
Artículo en Inglés | MEDLINE | ID: mdl-25750421

RESUMEN

Apple production depends on the fulfilment of a chilling requirement for bud dormancy release. Insufficient winter chilling results in irregular and suboptimal bud break in the spring, with negative impacts on apple yield. Trees from apple cultivars with contrasting chilling requirements for bud break were used to investigate the expression of the entire set of apple genes in response to chilling accumulation in the field and controlled conditions. Total RNA was analysed on the AryANE v.1.0 oligonucleotide microarray chip representing 57,000 apple genes. The data were tested for functional enrichment, and differential expression was confirmed by real-time PCR. The largest number of differentially expressed genes was found in samples treated with cold temperatures. Cold exposure mostly repressed expression of transcripts related to photosynthesis, and long-term cold exposure repressed flavonoid biosynthesis genes. Among the differentially expressed selected candidates, we identified genes whose annotations were related to the circadian clock, hormonal signalling, regulation of growth, and flower development. Two genes, annotated as FLOWERING LOCUS C-like and MADS AFFECTING FLOWERING, showed strong differential expression in several comparisons. One of these two genes was upregulated in most comparisons involving dormancy release, and this gene's chromosomal position co-localized with the confidence interval of a major quantitative trait locus for the timing of bud break. These results indicate that photosynthesis and auxin transport are major regulatory nodes of apple dormancy and unveil strong candidates for the control of bud dormancy.


Asunto(s)
Frío , Genes de Plantas , Malus/genética , Relojes Circadianos , Análisis por Conglomerados , Flavonoides/biosíntesis , Flores/genética , Flores/crecimiento & desarrollo , Perfilación de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Malus/crecimiento & desarrollo , Análisis de Secuencia por Matrices de Oligonucleótidos , Sitios de Carácter Cuantitativo , Estaciones del Año , Transducción de Señal
15.
Plant Cell ; 24(2): 463-81, 2012 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-22307851

RESUMEN

The conserved Target of Rapamycin (TOR) kinase forms high molecular mass complexes and is a major regulator of cellular adaptations to environmental cues. The Lethal with Sec Thirteen 8/G protein ß subunit-like (LST8/GßL) protein is a member of the TOR complexes, and two putative LST8 genes are present in Arabidopsis thaliana, of which only one (LST8-1) is significantly expressed. The Arabidopsis LST8-1 protein is able to complement yeast lst8 mutations and interacts with the TOR kinase. Mutations in the LST8-1 gene resulted in reduced vegetative growth and apical dominance with abnormal development of flowers. Mutant plants were also highly sensitive to long days and accumulated, like TOR RNA interference lines, higher amounts of starch and amino acids, including proline and glutamine, while showing reduced concentrations of inositol and raffinose. Accordingly, transcriptomic and enzymatic analyses revealed a higher expression of genes involved in nitrate assimilation when lst8-1 mutants were shifted to long days. The transcriptome of lst8-1 mutants in long days was found to share similarities with that of a myo-inositol 1 phosphate synthase mutant that is also sensitive to the extension of the light period. It thus appears that the LST8-1 protein has an important role in regulating amino acid accumulation and the synthesis of myo-inositol and raffinose during plant adaptation to long days.


Asunto(s)
Aclimatación/genética , Proteínas de Arabidopsis/metabolismo , Arabidopsis/genética , Flores/crecimiento & desarrollo , Fotoperiodo , Aminoácidos/metabolismo , Arabidopsis/crecimiento & desarrollo , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Regulación de la Expresión Génica de las Plantas , Metaboloma , Mutación , Fosfatidilinositol 3-Quinasas/metabolismo , Plantas Modificadas Genéticamente/genética , Plantas Modificadas Genéticamente/crecimiento & desarrollo , Plantas Modificadas Genéticamente/metabolismo , Interferencia de ARN , Transcriptoma
16.
Plant Cell ; 24(10): 4044-65, 2012 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-23054470

RESUMEN

The role of NADH-dependent glutamate dehydrogenase (GDH) was investigated by studying the physiological impact of a complete lack of enzyme activity in an Arabidopsis thaliana plant deficient in three genes encoding the enzyme. This study was conducted following the discovery that a third GDH gene is expressed in the mitochondria of the root companion cells, where all three active GDH enzyme proteins were shown to be present. A gdh1-2-3 triple mutant was constructed and exhibited major differences from the wild type in gene transcription and metabolite concentrations, and these differences appeared to originate in the roots. By placing the gdh triple mutant under continuous darkness for several days and comparing it to the wild type, the evidence strongly suggested that the main physiological function of NADH-GDH is to provide 2-oxoglutarate for the tricarboxylic acid cycle. The differences in key metabolites of the tricarboxylic acid cycle in the triple mutant versus the wild type indicated that, through metabolic processes operating mainly in roots, there was a strong impact on amino acid accumulation, in particular alanine, γ-aminobutyrate, and aspartate in both roots and leaves. These results are discussed in relation to the possible signaling and physiological functions of the enzyme at the interface of carbon and nitrogen metabolism.


Asunto(s)
Oxidorreductasas de Alcohol/fisiología , Proteínas de Arabidopsis/fisiología , Arabidopsis/enzimología , Carbono/metabolismo , Nitrógeno/metabolismo , Raíces de Plantas/metabolismo , Alanina/metabolismo , Oxidorreductasas de Alcohol/análisis , Oxidorreductasas de Alcohol/química , Oxidorreductasas de Alcohol/genética , Arabidopsis/genética , Proteínas de Arabidopsis/análisis , Proteínas de Arabidopsis/química , Proteínas de Arabidopsis/genética , Ácido Aspártico/metabolismo , Ciclo del Ácido Cítrico , Perfilación de la Expresión Génica , Ácidos Cetoglutáricos/metabolismo , Modelos Biológicos , Datos de Secuencia Molecular , Mutación , Hojas de la Planta/metabolismo , Transducción de Señal , Ácido gamma-Aminobutírico/metabolismo
17.
Plant J ; 73(5): 836-49, 2013 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-23181524

RESUMEN

Photorespiratory metabolism is essential in all oxygenic photosynthetic organisms. In plants, it is a highly compartmentalized pathway that involves chloroplasts, peroxisomes, mitochondria and the cytoplasm. The metabolic pathway itself is well characterized, and the enzymes required for its function have been identified. However, very little information is available on the transport proteins that catalyze the high metabolic flux between the involved compartments. Here we show that the A BOUT DE SOUFFLE (BOU) gene, which encodes a mitochondrial carrier, is involved in photorespiration in Arabidopsis. BOU was found to be co-expressed with photorespiratory genes in leaf tissues. The knockout mutant bou-2 showed the hallmarks of a photorespiratory growth phenotype, an elevated CO(2) compensation point, and excessive accumulation of glycine. Furthermore, degradation of the P-protein, a subunit of glycine decarboxylase, was demonstrated for bou-2, and is reflected in strongly reduced glycine decarboxylase activity. The photorespiration defect in bou-2 has dramatic consequences early in the seedling stage, which are highlighted by transcriptome studies. In bou-2 seedlings, as in shm1, another photorespiratory mutant, the shoot apical meristem organization is severely compromised. Cell divisions are arrested, leading to growth arrest at ambient CO(2) . Although the specific substrate for the BOU transporter protein remains elusive, we show that it is essential for the function of the photorespiratory metabolism. We hypothesize that BOU function is linked with glycine decarboxylase activity, and is required for normal apical meristems functioning in seedlings.


Asunto(s)
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Dióxido de Carbono/metabolismo , Glicina-Deshidrogenasa (Descarboxilante)/metabolismo , Proteínas de Transporte de Membrana/genética , Meristema/genética , Aminoácidos/análisis , Aminoácidos/metabolismo , Arabidopsis/citología , Arabidopsis/fisiología , Arabidopsis/efectos de la radiación , Proteínas de Arabidopsis/metabolismo , Respiración de la Célula , Perfilación de la Expresión Génica , Prueba de Complementación Genética , Glicina/metabolismo , Luz , Proteínas de Transporte de Membrana/metabolismo , Meristema/citología , Meristema/fisiología , Meristema/efectos de la radiación , Redes y Vías Metabólicas , Mitocondrias/metabolismo , Proteínas Mitocondriales/genética , Proteínas Mitocondriales/metabolismo , Mutación , Análisis de Secuencia por Matrices de Oligonucleótidos , Oxidación-Reducción , Fotosíntesis , Hojas de la Planta/citología , Hojas de la Planta/genética , Hojas de la Planta/fisiología , Hojas de la Planta/efectos de la radiación , Plantas Modificadas Genéticamente , Plantones/citología , Plantones/genética , Plantones/fisiología , Plantones/efectos de la radiación
18.
BMC Plant Biol ; 14: 375, 2014 Dec 31.
Artículo en Inglés | MEDLINE | ID: mdl-25551767

RESUMEN

BACKGROUND: Apple fruit mealiness is one of the most important textural problems that results from an undesirable ripening process during storage. This phenotype is characterized by textural deterioration described as soft, grainy and dry fruit. Despite several studies, little is known about mealiness development and the associated molecular events. In this study, we integrated phenotypic, microscopic, transcriptomic and biochemical analyses to gain insights into the molecular basis of mealiness development. RESULTS: Instrumental texture characterization allowed the refinement of the definition of apple mealiness. In parallel, a new and simple quantitative test to assess this phenotype was developed. CONCLUSIONS: These data support the role of PME in cell wall remodelling during apple fruit development and ripening and suggest a local action of these enzymes. Mealiness may partially result from qualitative and spatial variations of pectin microarchitecture rather than quantitative pectin differences, and these changes may occur early in fruit development. The specific MdPME2 gene highlighted in this study could be a good early marker of texture unfavourable trait in apple.


Asunto(s)
Hidrolasas de Éster Carboxílico/metabolismo , Frutas/enzimología , Frutas/metabolismo , Malus/enzimología , Malus/metabolismo , Frutas/crecimiento & desarrollo , Regulación de la Expresión Génica de las Plantas
19.
New Phytol ; 203(1): 287-99, 2014 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-24690119

RESUMEN

Characterizing the transcriptome of eukaryotic organisms is essential for studying gene regulation and its impact on phenotype. The realization that anti-sense (AS) and noncoding RNA transcription is pervasive in many genomes has emphasized our limited understanding of gene transcription and post-transcriptional regulation. Numerous mechanisms including convergent transcription, anti-correlated expression of sense and AS transcripts, and RNAi remain ill-defined. Here, we have combined microarray analysis and high-throughput sequencing of small RNAs (sRNAs) to unravel the complexity of transcriptional and potential post-transcriptional regulation in eight organs of apple (Malus × domestica). The percentage of AS transcript expression is higher than that identified in annual plants such as rice and Arabidopsis thaliana. Furthermore, we show that a majority of AS transcripts are transcribed beyond 3'UTR regions, and may cover a significant portion of the predicted sense transcripts. Finally we demonstrate at a genome-wide scale that anti-sense transcript expression is correlated with the presence of both short (21-23 nt) and long (> 30 nt) siRNAs, and that the sRNA coverage depth varies with the level of AS transcript expression. Our study provides a new insight on the functional role of anti-sense transcripts at the genome-wide level, and a new basis for the understanding of sRNA biogenesis in plants.


Asunto(s)
Malus/genética , Procesamiento Postranscripcional del ARN , ARN sin Sentido/genética , ARN Interferente Pequeño/genética , Transcripción Genética , Regulación de la Expresión Génica de las Plantas , Secuenciación de Nucleótidos de Alto Rendimiento , Análisis de Secuencia por Matrices de Oligonucleótidos , ARN de Planta/genética , Análisis de Secuencia de ARN
20.
Plant Cell ; 23(5): 1849-60, 2011 May.
Artículo en Inglés | MEDLINE | ID: mdl-21571951

RESUMEN

In plants, light represents an important environmental signal that triggers the production of photosynthetically active chloroplasts. This developmental switch is critical for plant survival because chlorophyll precursors that accumulate in darkness can be extremely destructive when illuminated. Thus, plants have evolved mechanisms to adaptively control plastid development during the transition into light. Here, we report that the gibberellin (GA)-regulated DELLA proteins play a crucial role in the formation of functional chloroplasts during deetiolation. We show that Arabidopsis thaliana DELLAs accumulating in etiolated cotyledons derepress chlorophyll and carotenoid biosynthetic pathways in the dark by repressing the transcriptional activity of the phytochrome-interacting factor proteins. Accordingly, dark-grown GA-deficient ga1-3 mutants (that accumulate DELLAs) display a similar gene expression pattern to wild-type seedlings grown in the light. Consistent with this, ga1-3 seedlings accumulate higher amounts of protochlorophyllide (a phototoxic chlorophyll precursor) in darkness but, surprisingly, are substantially more resistant to photooxidative damage following transfer into light. This is due to the DELLA-dependent upregulation of the photoprotective enzyme protochlorophyllide oxidoreductase (POR) in the dark. Our results emphasize the role of DELLAs in regulating the levels of POR, protochlorophyllide, and carotenoids in the dark and in protecting etiolated seedlings against photooxidative damage during initial light exposure.


Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/fisiología , Carotenoides/metabolismo , Clorofila/metabolismo , Oxidorreductasas actuantes sobre Donantes de Grupo CH-CH/metabolismo , Arabidopsis/enzimología , Arabidopsis/genética , Arabidopsis/efectos de la radiación , Proteínas de Arabidopsis/genética , Carotenoides/efectos de la radiación , Clorofila/efectos de la radiación , Cotiledón/genética , Cotiledón/fisiología , Cotiledón/efectos de la radiación , Oscuridad , Regulación de la Expresión Génica de las Plantas , Giberelinas/metabolismo , Luz , Análisis de Secuencia por Matrices de Oligonucleótidos , Estrés Oxidativo , Oxidorreductasas actuantes sobre Donantes de Grupo CH-CH/genética , Fotoblanqueo , Fitocromo/metabolismo , Protoclorofilida/metabolismo , Protoclorofilida/efectos de la radiación , Especies Reactivas de Oxígeno/metabolismo , Plantones/genética , Plantones/crecimiento & desarrollo , Plantones/efectos de la radiación , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Transcriptoma
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