RESUMEN
Photoperiods integrate with the circadian clock to coordinate gene expression rhythms and thus ensure plant fitness to the environment. Genome-wide characterization and comparison of rhythmic genes under different light conditions revealed delayed phase under constant darkness (DD) and reduced amplitude under constant light (LL) in rice. Interestingly, ChIP-seq and RNA-seq profiling of rhythmic genes exhibit synchronous circadian oscillation in H3K9ac modifications at their loci and long non-coding RNAs (lncRNAs) expression at proximal loci. To investigate how gene expression rhythm is regulated in rice, we profiled the open chromatin regions and transcription factor (TF) footprints by time-series ATAC-seq. Although open chromatin regions did not show circadian change, a significant number of TFs were identified to rhythmically associate with chromatin and drive gene expression in a time-dependent manner. Further transcriptional regulatory networks mapping uncovered significant correlation between core clock genes and transcription factors involved in light/temperature signaling. In situ Hi-C of ZT8-specific expressed genes displayed highly connected chromatin association at the same time, whereas this ZT8 chromatin connection network dissociates at ZT20, suggesting the circadian control of gene expression by dynamic spatial chromatin conformation. These findings together implicate the existence of a synchronization mechanism between circadian H3K9ac modifications, chromatin association of TF and gene expression, and provides insights into circadian dynamics of spatial chromatin conformation that associate with gene expression rhythms.
Asunto(s)
Ritmo Circadiano , Oryza , Cromatina/genética , Relojes Circadianos/genética , Ritmo Circadiano/genética , Epigenoma , Perfilación de la Expresión Génica , Oryza/genética , Oryza/fisiología , Factores de Transcripción/genéticaRESUMEN
Histone modification H3K27me3 is an important chromatin mark that plays vital roles in repressing expression of developmental genes. Here, we construct high-resolution 3D genome maps using long-read chromatin interaction analysis by paired-end tag sequencing (ChIA-PET) and characterize H3K27me3-associated chromatin interactions in an elite rice hybrid, Shanyou 63. We find that many H3K27me3-marked regions may function as silencer-like regulatory elements. The silencer-like elements can come into proximity with distal target genes via forming chromatin loops in 3D space of the nuclei, regulating gene silencing and plant traits. Natural and induced deletion of silencers upregulate expression of distal connected genes. Furthermore, we identify extensive allele-specific chromatin loops. We find that genetic variations alter allelic chromatin topology, thus modulating allelic gene imprinting in rice hybrids. In conclusion, the characterization of silencer-like regulatory elements and haplotype-resolved chromatin interaction maps provide insights into the understanding of molecular mechanisms underlying allelic gene silencing and plant trait controlling.
Asunto(s)
Cromatina , Oryza , Cromatina/metabolismo , Histonas/genética , Histonas/metabolismo , Oryza/genética , Haplotipos , Silenciador del GenRESUMEN
Lipid droplet (LD) in vegetative tissues has recently been implicated in environmental responses in plants, but its regulation and its function in stress tolerance are not well understood. Here, we identified a Membrane Occupation and Recognition Nexus 1 (MORN1) gene as a contributor to natural variations of stress tolerance through genome-wide association study in Arabidopsis thaliana. Characterization of its loss-of-function mutant and natural variants revealed that the MORN1 gene is a positive regulator of plant growth, disease resistance, cold tolerance, and heat tolerance. The MORN1 protein is associated with the Golgi and is also partly associated with LD. Protein truncations that disrupt these associations abolished the biological function of the MORN1 protein. Furthermore, the MORN1 gene is a positive regulator of LD abundance, and its role in LD number regulation and stress tolerance is highly linked. Therefore, this study identifies MORN1 as a positive regulator of LD abundance and a contributor to natural variations of stress tolerance. It implicates a potential involvement of Golgi in LD biogenesis and strongly suggests a contribution of LD to diverse processes of plant growth and stress responses.
Asunto(s)
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/metabolismo , Gotas Lipídicas/metabolismo , Estudio de Asociación del Genoma Completo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Aparato de Golgi/metabolismo , Regulación de la Expresión Génica de las Plantas/genética , Plantas Modificadas Genéticamente/metabolismo , Estrés Fisiológico/genéticaRESUMEN
Copper (Cu) and iron (Fe) are essential micronutrients that are toxic when accumulating in excess in cells. Thus, their uptake by roots is tightly regulated. While plants sense and respond to local Cu availability, the systemic regulation of Cu uptake has not been documented in contrast to local and systemic control of Fe uptake. Fe abundance in the phloem has been suggested to act systemically, regulating the expression of Fe uptake genes in roots. Consistently, shoot-to-root Fe signaling is disrupted in Arabidopsis thaliana mutants lacking the phloem companion cell-localized Fe transporter, OLIGOPEPTIDE TRANSPORTER 3 (AtOPT3). We report that AtOPT3 also transports Cu in heterologous systems and contributes to its delivery from sources to sinks in planta. The opt3 mutant contained less Cu in the phloem, was sensitive to Cu deficiency and mounted a transcriptional Cu deficiency response in roots and young leaves. Feeding the opt3 mutant and Cu- or Fe-deficient wild-type seedlings with Cu or Fe via the phloem in leaves downregulated the expression of both Cu- and Fe-deficiency marker genes in roots. These data suggest the existence of shoot-to-root Cu signaling, highlight the complexity of Cu/Fe interactions, and the role of AtOPT3 in fine-tuning root transcriptional responses to the plant Cu and Fe needs.
Asunto(s)
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/genética , Arabidopsis/metabolismo , Cobre , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Floema/genética , Floema/metabolismo , Homeostasis , Hierro/metabolismo , Plantas/metabolismo , Proteínas de Transporte de Membrana/metabolismoRESUMEN
Plant photoperiodic growth is coordinated by interactions between circadian clock and light signaling networks. How post-translational modifications of clock proteins affect these interactions to mediate rhythmic growth remains unclear. Here, we identify five phosphorylation sites in the Arabidopsis core clock protein TIMING OF CAB EXPRESSION 1 (TOC1) which when mutated to alanine eliminate detectable phosphorylation. The TOC1 phospho-mutant fails to fully rescue the clock, growth, and flowering phenotypes of the toc1 mutant. Further, the TOC1 phospho-mutant shows advanced phase, a faster degradation rate, reduced interactions with PHYTOCHROME-INTERACTING FACTOR 3 (PIF3) and HISTONE DEACETYLASE 15 (HDA15), and poor binding at pre-dawn hypocotyl growth-related genes (PHGs), leading to a net de-repression of hypocotyl growth. NUCLEAR FACTOR Y subunits B and C (NF-YB/C) stabilize TOC1 at target promoters, and this novel trimeric complex (NF-TOC1) acts as a transcriptional co-repressor with HDA15 to inhibit PIF-mediated hypocotyl elongation. Collectively, we identify a molecular mechanism suggesting how phosphorylation of TOC1 alters its phase, stability, and physical interactions with co-regulators to precisely phase PHG expression to control photoperiodic hypocotyl growth.
Asunto(s)
Proteínas de Arabidopsis/genética , Arabidopsis/crecimiento & desarrollo , Factor de Unión a CCAAT/metabolismo , Mutación , Factores de Transcripción/genética , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/química , Proteínas de Arabidopsis/metabolismo , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/metabolismo , Regulación de la Expresión Génica de las Plantas , Histona Desacetilasas/metabolismo , Hipocótilo/crecimiento & desarrollo , Hipocótilo/metabolismo , Fosforilación , Proteolisis , Transducción de Señal , Factores de Transcripción/química , Factores de Transcripción/metabolismoRESUMEN
The molecular components of the circadian system possess the interesting feature of acting together to create a self-sustaining oscillator, while at the same time acting individually, and in complexes, to confer phase-specific circadian control over a wide range of physiological and developmental outputs. This means that many circadian oscillator proteins are simultaneously also part of the circadian output pathway. Most studies have focused on transcriptional control of circadian rhythms, but work in plants and metazoans has shown the importance of post-transcriptional and post-translational processes within the circadian system. Here we highlight recent work describing post-translational mechanisms that impact both the function of the oscillator and the clock-controlled outputs.
Asunto(s)
Relojes Circadianos/fisiología , Ritmo Circadiano/fisiología , Regulación de la Expresión Génica de las Plantas/fisiología , Proteínas de Plantas/biosíntesis , Plantas/metabolismo , Procesamiento Proteico-Postraduccional/fisiología , Proteínas de Plantas/genética , Plantas/genéticaRESUMEN
Nuclear pore complexes (NPCs) are main channels controlling nucleocytoplasmic transport and are composed of approximately 30 nucleoporins (NUPs). Emerging evidence suggests that some NUP genes have specialized functions that challenge the traditional view of NPCs as structures of uniform composition. Here, we analysed the role of six outer-ring components of NPC at normal and warm growth temperatures by examining their loss-of-function mutants in Arabidopsis thaliana. All six NUP subunits, NUP85, NUP96, NUP 133, NUP 160, SEH1 and HOS1, have a non-redundant temperature-influenced function in one or more of the processes, including rosette growth, leaf architecture and intracellular immune receptor-mediated disease resistance. At the molecular level, NUP85 and NUP133 are required for mRNA export only at warm temperature and play a larger role in the localization of transcription factor at warm temperature. In addition, NUP96 and HOS1 are essential for the expression of high temperature-responsive genes, which is correlated with their larger activity in facilitating nuclear accumulation of the transcription factor PIF4 at warm temperature. Our results show that subunits of NPC have differential roles at different temperatures, suggesting the existence of temperature-influenced NPC complexes and activities.
Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/crecimiento & desarrollo , Arabidopsis/inmunología , Proteínas de Complejo Poro Nuclear/metabolismo , Desarrollo de la Planta , Inmunidad de la Planta , Temperatura , Arabidopsis/genética , Arabidopsis/microbiología , Núcleo Celular/metabolismo , Regulación de la Expresión Génica de las Plantas , Genes de Plantas , Mutación con Pérdida de Función , Fenotipo , Transporte de ARN/genética , ARN Mensajero/genética , ARN Mensajero/metabolismo , Fracciones Subcelulares/metabolismo , Transcripción Genética , VirulenciaRESUMEN
Temperature has a large impact on plant immune responses. Earlier studies identified intracellular immune receptor nucleotide-binding leucine-rich repeat (NLR) genes and salicylic acid (SA) as targets of high-temperature inhibition of plant immunity. Here, we report that moderately low temperature enhances immunity to the bacterial pathogen Pseudomonas syringae in Arabidopsis (Arabidopsis thaliana). This enhancement is dependent on SA signaling and is accompanied by up-regulation of multiple SA biosynthesis and signaling genes at lower temperature. SA signaling is repressed by jasmonic acid and ethylene at both normal and low temperatures. The inhibition of SA biosynthesis by ethylene, while mainly through ISOCHORISMATE SYNTHASE1/SALICYLIC ACID-INDUCTION DEFICIENT2 (ICS1/SID2) at normal temperature, is through ENHANCED DISEASE SUSCEPTIBILITY5 (EDS5)/SID1, ICS2, and ICS1/SID2 at lower temperature. The repression by ethylene is mediated by a direct regulation of the ethylene response transcription factor ETHYLENE INSENSITIVE3 (EIN3) on multiple SA biosynthesis and signaling genes. Thus, low temperature enhances the SA pathway to promote immunity and at the same time uses ethylene to repress multiple SA regulators to achieve fine-tuned immune responses.
Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Etilenos/farmacología , Inmunidad de la Planta/fisiología , Ácido Salicílico/metabolismo , Arabidopsis/efectos de los fármacos , Arabidopsis/genética , Arabidopsis/microbiología , Proteínas de Arabidopsis/genética , Ciclopentanos/farmacología , Transferasas Intramoleculares/genética , Transferasas Intramoleculares/metabolismo , Oxilipinas/farmacología , Inmunidad de la Planta/genética , Plantas Modificadas Genéticamente/efectos de los fármacos , Plantas Modificadas Genéticamente/genética , Plantas Modificadas Genéticamente/metabolismo , Plantas Modificadas Genéticamente/microbiología , Pseudomonas syringae/patogenicidad , TemperaturaRESUMEN
Stomatal closure defense and apoplastic defense are two major immunity mechanisms restricting the entry and propagation of microbe pathogens in plants. Surprisingly, activation of plant intracellular immune receptor NLR genes, while enhancing whole plant disease resistance, was sometimes linked to a defective stomatal defense in autoimmune mutants. Here we report the use of high temperature and genetic chimera to investigate the inter-dependence of stomatal and apoplastic defenses in autoimmunity. High temperature inhibits both stomatal and apoplastic defenses in the wild type, suppresses constitutive apoplastic defense responses and rescues the deficiency of stomatal closure response in autoimmune mutants. Chimeric plants have been generated to activate NLR only in guard cells or the non-guard cells. NLR activation in guard cells inhibits stomatal closure defense response in a cell autonomous manner likely through repressing ABA responses. At the same time, it leads to increased whole plant resistance accompanied by a slight increase in apoplastic defense. In addition, NLR activation in both guard and non-guard cells affects stomatal aperture and water potential. This study thus reveals that NLR activation has a differential effect on immunity in a cell type specific matter, which adds another layer of immune regulation with spatial information.
Asunto(s)
Arabidopsis/inmunología , Resistencia a la Enfermedad/genética , Proteínas NLR/metabolismo , Estomas de Plantas/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Autoinmunidad/genética , Autoinmunidad/inmunología , Quimera/genética , Regulación de la Expresión Génica de las Plantas , Calor , Receptores Inmunológicos/metabolismoRESUMEN
Plant cells have multiple plasma membrane (PM)-localized calcium ATPases (ACAs) pumping calcium ions out of the cytosol. Although the involvement of some of these ACAs in plant growth and immunity has been reported, their individual and combined functions have not been fully examined. Here, we analysed the effects of single and combined mutations of four ACA genes, ACA8, ACA10, ACA12, and ACA13, in a number of processes. We found that these four genes had both overlapping and differential involvements in vegetative growth, inflorescence growth, seeds setting, disease resistance and stomatal movement. Disruption of any of these four genes reduces seed setting, indicating their contribution to the overall fitness of the plants. While ACA10 and ACA8 play major roles in vegetative growth and immunity, ACA13 and ACA12 are also involved in these processes especially when the function of ACA10 and/or ACA8 is compromised. The loss of ACA13 and ACA10 function in combination with a reduction in function of ACA8 leads to seedling death at bolting, revealing the essential role of their collective function in plant growth. Taken together, this study indicates a highly tuned calcium system involving these PM-localized calcium pumps in plant growth and environmental responses.
Asunto(s)
Proteínas de Arabidopsis/genética , Arabidopsis/fisiología , ATPasas Transportadoras de Calcio de la Membrana Plasmática/genética , Arabidopsis/genética , Arabidopsis/crecimiento & desarrollo , Proteínas de Arabidopsis/metabolismo , Regulación de la Expresión Génica de las Plantas , Mutación , ATPasas Transportadoras de Calcio de la Membrana Plasmática/metabolismoRESUMEN
A deficiency of the micronutrient copper (Cu) leads to infertility and grain/seed yield reduction in plants. How Cu affects fertility, which reproductive structures require Cu, and which transcriptional networks coordinate Cu delivery to reproductive organs is poorly understood. Using RNA-seq analysis, we showed that the expression of a gene encoding a novel transcription factor, CITF1 (Cu-DEFICIENCY INDUCED TRANSCRIPTION FACTOR1), was strongly upregulated in Arabidopsis thaliana flowers subjected to Cu deficiency. We demonstrated that CITF1 regulates Cu uptake into roots and delivery to flowers and is required for normal plant growth under Cu deficiency. CITF1 acts together with a master regulator of copper homeostasis, SPL7 (SQUAMOSA PROMOTER BINDING PROTEIN LIKE7), and the function of both is required for Cu delivery to anthers and pollen fertility. We also found that Cu deficiency upregulates the expression of jasmonic acid (JA) biosynthetic genes in flowers and increases endogenous JA accumulation in leaves. These effects are controlled in part by CITF1 and SPL7. Finally, we show that JA regulates CITF1 expression and that the JA biosynthetic mutant lacking the CITF1- and SPL7-regulated genes, LOX3 and LOX4, is sensitive to Cu deficiency. Together, our data show that CITF1 and SPL7 regulate Cu uptake and delivery to anthers, thereby influencing fertility, and highlight the relationship between Cu homeostasis, CITF1, SPL7, and the JA metabolic pathway.
Asunto(s)
Proteínas de Arabidopsis/metabolismo , Factores de Transcripción Básicos con Cremalleras de Leucinas y Motivos Hélice-Asa-Hélice/genética , Cobre/farmacología , Ciclopentanos/metabolismo , Proteínas de Unión al ADN/metabolismo , Fertilidad/fisiología , Oxilipinas/metabolismo , Polen/fisiología , Factores de Transcripción/metabolismo , Arabidopsis/genética , Arabidopsis/crecimiento & desarrollo , Proteínas de Arabidopsis/genética , Factores de Transcripción Básicos con Cremalleras de Leucinas y Motivos Hélice-Asa-Hélice/metabolismo , Vías Biosintéticas/genética , Núcleo Celular/efectos de los fármacos , Núcleo Celular/metabolismo , Cobre/deficiencia , Ciclopentanos/farmacología , Proteínas de Unión al ADN/genética , Fertilidad/efectos de los fármacos , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Homeostasis , Modelos Biológicos , Mutación/genética , Oxilipinas/farmacología , Fenotipo , Polen/efectos de los fármacos , Transporte de Proteínas/efectos de los fármacos , Protoplastos/efectos de los fármacos , Protoplastos/metabolismo , Factores de Transcripción/genética , Transcripción Genética/efectos de los fármacos , Transcriptoma/genética , Regulación hacia Arriba/efectos de los fármacos , Regulación hacia Arriba/genéticaRESUMEN
Calcium signaling is essential for environmental responses including immune responses. Here, we provide evidence that the evolutionarily conserved protein BONZAI1 (BON1) functions together with autoinhibited calcium ATPase10 (ACA10) and ACA8 to regulate calcium signals in Arabidopsis. BON1 is a plasma membrane localized protein that negatively regulates the expression of immune receptor genes and positively regulates stomatal closure. We found that BON1 interacts with the autoinhibitory domains of ACA10 and ACA8, and the aca10 loss-of-function (LOF) mutants have an autoimmune phenotype similar to that of the bon1 LOF mutants. Genetic evidences indicate that BON1 positively regulates the activities of ACA10 and ACA8. Consistent with this idea, the steady level of calcium concentration is increased in both aca10 and bon1 mutants. Most strikingly, cytosolic calcium oscillation imposed by external calcium treatment was altered in aca10, aca8, and bon1 mutants in guard cells. In addition, calcium- and pathogen-induced stomatal closure was compromised in the aca10 and bon1 mutants. Taken together, this study indicates that ACA10/8 and BON1 physically interact on plasma membrane and function in the generation of cytosol calcium signatures that are critical for stomatal movement and impact plant immunity.
Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/genética , Señalización del Calcio , ATPasas Transportadoras de Calcio/metabolismo , Calcio/metabolismo , Proteínas Portadoras/metabolismo , Proteínas de la Membrana/metabolismo , Arabidopsis/inmunología , Arabidopsis/fisiología , Proteínas de Arabidopsis/genética , Proteínas de Unión al Calcio , ATPasas Transportadoras de Calcio/genética , Proteínas Portadoras/genética , Membrana Celular/metabolismo , Citosol/metabolismo , Genes Reporteros , Homeostasis , Mutación con Pérdida de Función , Proteínas de la Membrana/genética , Inmunidad de la Planta , Estomas de Plantas/genética , Estomas de Plantas/inmunología , Estomas de Plantas/fisiologíaRESUMEN
Urea is an important source of nitrogen (N) for the growth and development of plants. It occurs naturally in soils, is the major N source in agricultural fertilizers and is an important N metabolite in plants. Therefore, the identification and characterization of urea transporters in higher plants is important for the fundamental understanding of urea-based N nutrition in plants and for designing novel strategies for improving the N-use efficiency of urea based-fertilizers. Progress in this area, however, is hampered due to scarce knowledge of plant urea transporters. From what is known, urea uptake from the soil into plant roots is mediated by two types of transporters: the major intrinsic proteins (MIPs) and the DUR3 orthologs, mediating low- and high-affinity urea transport, respectively. Here we characterized a MIP family member from Cucumis sativus, CsNIP2;1, with regard to its contribution to urea transport. We show that CsNIP2;1 is a plasma membrane transporter that mediates pH-dependent urea uptake when expressed in yeast. We also found that ectopic expression of CsNIP2;1 improves growth of wild-type Arabidopsis thaliana and rescues growth and development of the atdur3-3 mutant on medium with urea as the sole N source. In addition, CsNIP2;1 is transcriptionally up-regulated by N deficiency, urea and NO3 (-). These data and results from the analyses of the pattern of CsNIP2;1 expression in A. thaliana and cucumber suggest that CsNIP2;1 might be involved in multiple steps of urea-based N nutrition, including urea uptake and internal transport during N remobilization throughout seed germination and N delivery to developing tissues.
Asunto(s)
Arabidopsis/genética , Membrana Celular/metabolismo , Cucumis sativus/metabolismo , Proteínas de Transporte de Membrana/metabolismo , Proteínas de Plantas/metabolismo , Saccharomyces cerevisiae/metabolismo , Urea/metabolismo , Secuencia de Aminoácidos , Arabidopsis/efectos de los fármacos , Arabidopsis/crecimiento & desarrollo , Membrana Celular/efectos de los fármacos , Cucumis sativus/efectos de los fármacos , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Prueba de Complementación Genética , Glicerol/farmacología , Proteínas de Transporte de Membrana/química , Mutación/genética , Nitratos/metabolismo , Nitrógeno/farmacología , Proteínas de Plantas/química , Raíces de Plantas/efectos de los fármacos , Raíces de Plantas/metabolismo , Plantas Modificadas Genéticamente , Transporte de Proteínas/efectos de los fármacos , Protoplastos/efectos de los fármacos , Protoplastos/metabolismo , Transformación Genética/efectos de los fármacos , Regulación hacia Arriba/efectos de los fármacosRESUMEN
The protoplast transient assay system has been widely used for rapid functional analyses of genes using cellular and biochemical approaches. This system has been increasingly employed for functional genetic studies using double-stranded (ds) RNA interference (RNAi). Here, we describe a modified procedure for the isolation of protoplasts from leaf mesophyll cells of 14-day-old Arabidopsis thaliana. This modification significantly simplifies and speeds up functional studies without compromising the yield and the viability of protoplasts. We also present the procedure for the isolation and transfection of protoplasts from mesophyll cells of an emerging model grass species, Brachypodium distachyon. Further, we detail procedures for RNAi-based functional studies of genes using transient expression of in vitro synthesized dsRNA in protoplasts.
Asunto(s)
Expresión Génica , Protoplastos/metabolismo , Transfección , Arabidopsis/genética , Brachypodium/genética , Regulación de la Expresión Génica de las Plantas , Células del Mesófilo/metabolismo , Interferencia de ARN , ARN Bicatenario/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Transfección/métodosRESUMEN
Copper (Cu) is an essential micronutrient that performs a remarkable array of functions in plants including photosynthesis, cell wall remodeling, flowering, and seed set. Of the world's major cereal crops, wheat, barley, and oat are the most sensitive to Cu deficiency. Cu deficient soils include alkaline soils, which occupy approximately 30% of the world's arable lands, and organic soils that occupy an estimated 19% of arable land in Europe. We used Brachypodium distachyon (brachypodium) as a proxy for wheat and other grain cereals to initiate analyses of the molecular mechanisms underlying their increased susceptibility to Cu deficiency. In this report, we focus on members of the CTR/COPT family of Cu transporters because their homologs in A. thaliana are transcriptionally upregulated in Cu-limited conditions and are involved either in Cu uptake from soils into epidermal cells in the root, or long-distance transport and distribution of Cu in photosynthetic tissues. We found that of five COPT proteins in brachypodium, BdCOPT3, and BdCOPT4 localize to the plasma membrane and are transcriptionally upregulated in roots and leaves by Cu deficiency. We also found that BdCOPT3, BdCOPT4, and BdCOPT5 confer low affinity Cu transport, in contrast to their counterparts in A. thaliana that confer high affinity Cu transport. These data suggest that increased sensitivity to Cu deficiency in some grass species may arise from lower efficiency and, possibly, other properties of components of Cu uptake and tissue partitioning systems and reinforce the importance of using brachypodium as a model for the comprehensive analyses of Cu homeostasis in cereal crops.
RESUMEN
Copper (Cu) homeostasis in plants is maintained by at least two mechanisms: (1) the miRNA-dependent reallocation of intracellular Cu among major Cu-enzymes and important energy-related functions; (2) the regulation of the expression of Cu transporters including members of the CTR/COPT family. These events are controlled by the transcription factor SPL7 in Arabidopsis thaliana. Cadmium (Cd), on the other hand, is a non-essential and a highly toxic metal that interferes with homeostasis of essential elements by competing for cellular binding sites. Whether Cd affects Cu homeostasis in plants is unknown. We found that Cd stimulates Cu accumulation in roots of A. thaliana and increases mRNA expression of three plasma membrane-localized Cu uptake transporters, COPT1, COPT2 and COPT6. Further analysis of Cd sensitivity of single and triple copt1copt2copt6 mutants, and transgenic plants ectopically expressing COPT6 suggested that Cu uptake is an essential component of Cd resistance in A. thaliana. Analysis of the contribution of the SPL7-dependent pathway to Cd-induced expression of COPT1, COPT2 and COPT6 showed that it occurs, in part, through mimicking the SPL7-dependent transcriptional Cu deficiency response. This response also involves components of the Cu reallocation system, miRNA398, FSD1, CSD1 and CSD2. Furthermore, seedlings of the spl7-1 mutant accumulate up to 2-fold less Cu in roots than the wild-type, are hypersensitive to Cd, and are more sensitive to Cd than the triple copt1copt2copt6 mutant. Together these data show that exposure to excess Cd triggers SPL7-dependent Cu deficiency responses that include Cu uptake and reallocation that are required for basal Cd tolerance in A. thaliana.
Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Cadmio/farmacología , Cobre/metabolismo , Proteínas de Unión al ADN/metabolismo , Factores de Transcripción/metabolismo , Adaptación Fisiológica/efectos de los fármacos , Adaptación Fisiológica/genética , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Proteínas de Transporte de Catión/genética , Proteínas de Transporte de Catión/metabolismo , Membrana Celular/metabolismo , Transportador de Cobre 1 , Proteínas de Unión al ADN/genética , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Prueba de Complementación Genética , Homeostasis/efectos de los fármacos , Proteínas de Transporte de Membrana/genética , Proteínas de Transporte de Membrana/metabolismo , MicroARNs/genética , Microscopía Fluorescente , Modelos Genéticos , Mutación , Raíces de Plantas/genética , Raíces de Plantas/metabolismo , Brotes de la Planta/genética , Brotes de la Planta/metabolismo , Plantas Modificadas Genéticamente , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Proteínas SLC31 , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/crecimiento & desarrollo , Saccharomyces cerevisiae/metabolismo , Superóxido Dismutasa/genética , Superóxido Dismutasa/metabolismo , Factores de Transcripción/genéticaRESUMEN
Submergence can severely affect the growth of perennial grasses. The variations in growth and the physiological responses of perennial grass germplasm to submergence stress are not well understood. The objective of this study was to characterize the responses of diverse perennial ryegrass accessions to submergence and their recovery following de-submergence. One hundred globally collected perennial ryegrass accessions were submerged for 7d followed by 7d of recovery in two experiments (Exp 1 and Exp 2), respectively. Compared to the pattern of the controls, the overall distribution in leaf color, chlorophyll fluorescence, plant height (HT), and growth rate (GR) shifted toward a high frequency of lower values under submergence in both experiments. The accessions were generally grouped into three types: fast growth with maintenance of color (escape, T1), slow growth with maintenance of color (quiescence, T2), and slow growth with loss of color (susceptible, ST). Under submergence, T1 had higher HT and GR than the other two groups except for GR of T2 in Exp 2 and had higher water-soluble carbohydrate (WSC) and fructan concentrations, as well as fructan to WSC ratio, than ST in Exp 1. Recovery of HT and GR were generally close to that of the control level except for HT of ST in Exp 2, but the carbohydrates fully recovered in all types of plants after 7d of de-submergence. Differential responses of perennial ryegrass accessions to submergence are useful in creating more tolerant materials and in further characterizing physiological and molecular mechanisms of submergence tolerance.