RESUMEN
Phosphorus is an essential macronutrient for plant growth and development. In response to phosphate (Pi) deficiency, plants rapidly produce a substitutive amount of root hairs; however, the mechanisms underlying Pi supply for root hair growth remain unclear. Here, we observed that soybean (Glycine max) plants maintain a consistent level of Pi within root hairs even under external Pi deficiency. We therefore investigated the role of vacuole-stored Pi, a major Pi reservoir in plant cells, in supporting root hair growth under Pi-deficient conditions. Our findings indicated that two vacuolar Pi efflux (VPE) transporters, GmVPE1 and GmVPE2, remobilize vacuolar stored Pi to sustain cytosolic Pi content in root hair cells. Genetic analysis showed that double mutants of GmVPE1 and GmVPE2 exhibited reduced root hair growth under low Pi conditions. Moreover, GmVPE1 and GmVPE2 were highly expressed in root hairs, with their expression levels significantly upregulated by low Pi treatment. Further analysis revealed that GmRSL2 (ROOT HAIR DEFECTIVE 6-like 2), a transcription factor involved in root hair morphogenesis, directly binds to the promoter regions of GmVPE1 and GmVPE2, and promotes their expressions under low Pi conditions. Additionally, mutants lacking both GmRSL2 and its homolog GmRSL3 exhibited impaired root hair growth under low Pi stress, which was rescued by overexpressing either GmVPE1 or GmVPE2. Taken together, our study has identified a module comprising vacuolar Pi exporters and transcription factors responsible for remobilizing vacuolar Pi to support root hair growth in response to Pi deficiency in soybean.
RESUMEN
The aleurone layer in cereal grains acts as a major reservoir of essential mineral nutrients, significantly influencing seed germination. However, the molecular mechanism underlying the redistribution of nutrients from the aleurone layer in the germinating seed is still not well understood. Here, in rice, we identified a plasma membrane (PM) localized magnesium transporter, MAGNESIUM RELEASE TRANSPORTER 3 (MGR3), is critical for seed germination. OsMGR3 is predominantly expressed in the aleurone layer cells of endosperm, facilitating magnesium remobilization during germination. Non-invasive Micro-test Technology assay data demonstrated that the loss-of-function of OsMGR3 restrained magnesium efflux from the aleurone layer. In the embryo/endosperm grafting experiment, we observed that the mutation of OsMGR3 in the aleurone layer suppressed the growth and differentiation of the embryo during germination. Furthermore, magnesium fluorescence imaging revealed the osmgr3 mutant seeds showed impaired exportation of aleurone layer-stored magnesium to the embryo, consequently delaying germination. Importantly, we discovered that disrupting OsMGR3 could inhibit pre-harvest sprouting without affecting rice yield and quality. Therefore, the magnesium efflux transporter OsMGR3 in the aleurone layer represents a promising genetic target for future agronomic trait improvement.
Asunto(s)
Membrana Celular , Germinación , Magnesio , Oryza , Proteínas de Plantas , Semillas , Oryza/genética , Oryza/metabolismo , Oryza/crecimiento & desarrollo , Oryza/fisiología , Magnesio/metabolismo , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Semillas/genética , Semillas/metabolismo , Semillas/crecimiento & desarrollo , Membrana Celular/metabolismo , Regulación de la Expresión Génica de las Plantas , Endospermo/metabolismo , Endospermo/genética , MutaciónRESUMEN
The plant genome produces an extremely large collection of long noncoding RNAs (lncRNAs) that are generally expressed in a context-specific manner and have pivotal roles in regulation of diverse biological processes. Here, we mapped the transcriptional heterogeneity of lncRNAs and their associated gene regulatory networks at single-cell resolution. We generated a comprehensive cell atlas at the whole-organism level by integrative analysis of 28 published single-cell RNA sequencing (scRNA-seq) datasets from juvenile Arabidopsis seedlings. We then provided an in-depth analysis of cell-type-related lncRNA signatures that show expression patterns consistent with canonical protein-coding gene markers. We further demonstrated that the cell-type-specific expression of lncRNAs largely explains their tissue specificity. In addition, we predicted gene regulatory networks on the basis of motif enrichment and co-expression analysis of lncRNAs and mRNAs, and we identified putative transcription factors orchestrating cell-type-specific expression of lncRNAs. The analysis results are available at the single-cell-based plant lncRNA atlas database (scPLAD; https://biobigdata.nju.edu.cn/scPLAD/). Overall, this work demonstrates the power of integrative single-cell data analysis applied to plant lncRNA biology and provides fundamental insights into lncRNA expression specificity and associated gene regulation.
Asunto(s)
Arabidopsis , ARN Largo no Codificante , Redes Reguladoras de Genes , ARN Largo no Codificante/genética , Arabidopsis/genética , Análisis de Expresión Génica de una Sola Célula , Regulación de la Expresión GénicaRESUMEN
Phosphorus (P) is an indispensable nutrient for seed germination, but the seeds always store excessive P over demand. High-P seeds of feeding crops lead to environmental and nutrition issues, because phytic acid (PA), the major form of P in seeds, cannot be digested by mono-gastric animals. Therefore, reduction of P level in seeds has become an imperative task in agriculture. Our study here suggested that both VPT1 and VPT3, two vacuolar phosphate transporters responsible for vacuolar Pi sequestration, were downregulated in leaves during the flowering stage, which led to less Pi accumulated in leaves and more Pi allocated to reproductive organs, and thus high-P containing seeds. To reduce the total P content in seeds, we genetically regulated VPT1 during the flowering stage and found that overexpression of VPT1 in leaves could reduce P content in seeds without affecting the production and seed vigor. Therefore, our finding provides a potential strategy to reduce the P level of the seeds to prevent the nutrition over-accumulation pollution.
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Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Semillas/genética , Semillas/metabolismo , Fósforo , Proteínas de Transporte de Fosfato/genética , Plantas Modificadas Genéticamente/metabolismoRESUMEN
Excess phosphate (Pi) is stored into the vacuole through Pi transporters so that cytoplasmic Pi levels remain stable in plant cells. We hypothesized that the vacuolar Pi transporters may harbor a Pi-sensing mechanism so that they are activated to deliver Pi into the vacuole only when cytosolic Pi reaches a threshold high level. We tested this hypothesis using Vacuolar Phosphate Transporter 1 (VPT1), a SPX domain-containing vacuolar Pi transporter, as a model. Recent studies have defined SPX as a Pi-sensing module that binds inositol polyphosphate signaling molecules (InsPs) produced at high cellular Pi status. We showed here that Pi-deficient conditions or mutation of the SPX domain severely impaired the transport activity of VPT1. We further identified an auto-inhibitory domain in VPT1 that suppresses its transport activity. Taking together the results from detailed structure-function analyses, our study suggests that VPT1 is in the auto-inhibitory state when Pi status is low, whereas at high cellular Pi status InsPs are produced and bind SPX domain to switch on VPT1 activity to deliver Pi into the vacuole. This thus provides an auto-regulatory mechanism for VPT1-mediated Pi sensing and homeostasis in plant cells.
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Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Homeostasis , Inositol , Proteínas de Transporte de Membrana/metabolismo , Proteínas de Transporte de Fosfato/genética , Fosfatos/metabolismo , Polifosfatos/metabolismo , Vacuolas/metabolismoRESUMEN
Magnesium (Mg2+ ) serves as a cofactor for a number of photosynthetic enzymes in the chloroplast, and is the central atom of the Chl molecule. However, little is known about the molecular mechanism of Mg2+ transport across the chloroplast envelope. Here, we report the functional characterization of two transport proteins in Arabidopsis: Magnesium Release 8 (MGR8) and MGR9, of the ACDP/CNNM family, which is evolutionarily conserved across all lineages of living organisms. Both MGR8 and MGR9 genes were expressed ubiquitously, and their encoded proteins were localized in the inner envelope of chloroplasts. Mutations of MGR8 and MGR9 together, but neither of them alone, resulted in albino ovules and chlorotic seedlings. Further analysis revealed severe defects in thylakoid biogenesis and assembly of photosynthetic complexes in the double mutant. Both MGR8 and MGR9 functionally complemented the growth of the Salmonella typhimurium mutant strain MM281, which lacks Mg2+ uptake capacity. The embryonic and early seedling defects of the mgr8/mgr9 double mutant were rescued by the expression of MGR9 under the embryo-specific ABI3 promoter. The partially rescued mutant plants were hypersensitive to Mg2+ deficient conditions and contained less Mg2+ in their chloroplasts than wild-type plants. Taken together, we conclude that MGR8 and MGR9 serve as Mg2+ transporters and are responsible for chloroplast Mg2+ uptake.
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Proteínas de Arabidopsis , Arabidopsis , Proteínas del Complejo del Centro de Reacción Fotosintética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Cloroplastos/metabolismo , Magnesio/metabolismo , Proteínas de Transporte de Membrana/metabolismo , Mutación/genética , Proteínas del Complejo del Centro de Reacción Fotosintética/genética , Plantones/metabolismo , Tilacoides/metabolismoRESUMEN
Nutrient antagonism typically refers to the fact that too high a concentration of one nutrient inhibits the absorption of another nutrient. In plants, Ca2+ (Calcium) and Mg2+ (Magnesium) are the two most abundant divalent cations, which are known to have antagonistic interactions. Hence, maintaining their homeostasis is crucial for plant growth and development. In this study, we showed that MTP10 (Metal Tolerance Protein 10) is an important regulator for maintaining homeostasis of Mg and Ca in Arabidopsis. The mtp10 mutant displayed severe growth retardation in the presence of excess Mg2+, to which the addition of Ca2+ was able to rescue the phenotype of mtp10 mutant. Additionally, the deficiency of Ca2+ in the culture medium accelerated the high-Mg sensitivity of the mtp10 mutant. The yeast complementation assay suggested that AtMTP10 had no Ca2+ transport activity. And the ICP-MS data further confirmed the antagonistic relationship between Ca2+ and Mg2+, with the addition of Ca2+ reducing the excessive accumulation of Mg2+ and high-Mg inhibiting the uptake of Ca2+. We conclude that the Arabidopsis MTP10 is essential for the regulation of Mg and Ca homeostasis.
Asunto(s)
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Calcio/metabolismo , Regulación de la Expresión Génica de las Plantas , Homeostasis , Magnesio/metabolismoRESUMEN
Magnesium (Mg2+), an essential structural component of chlorophyll, is absorbed from the soil by roots and transported to shoots to support photosynthesis in plants. However, the molecular mechanisms underlying root-to-shoot Mg2+ translocation remain largely unknown. We describe here the identification of four plasma membrane (PM)-localized transporters, named Mg2+ release transporters (MGRs), that are critical for root-to-shoot Mg transport in Arabidopsis. Functional complementation assays in a Mg2+-uptake-deficient bacterial strain confirmed that these MGRs conduct Mg2+ transport. PM-localized MGRs (MGR4, MGR5, MGR6, and MGR7) were expressed primarily in root stellar cells and participated in the xylem loading step of the long-distance Mg2+ transport process. In particular, MGR4 and MGR6 played a major role in shoot Mg homeostasis, as their loss-of-function mutants were hypersensitive to low Mg2+ but tolerant to high Mg2+ conditions. Reciprocal grafting analysis further demonstrated that MGR4 functions in the root to determine shoot Mg2+ accumulation and physiological phenotypes caused by both low- and high-Mg2+ stress. Taken together, our study has identified the long-sought transporters responsible for root-to-shoot Mg2+ translocation in plants.
Asunto(s)
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Transporte Biológico , Membrana Celular/metabolismo , Proteínas de Transporte de Membrana/metabolismo , Raíces de Plantas/metabolismo , Brotes de la Planta/metabolismo , Xilema/metabolismoRESUMEN
Magnesium (Mg2+) is an essential nutrient for all life forms. In fungal and plant cells, the majority of Mg2+ is stored in the vacuole but mechanisms for Mg2+ transport into the vacuolar store are not fully understood. Here we demonstrate that members of ancient conserved domain proteins (ACDPs) from Saccharomyces cerevisiae and Arabidopsis thaliana function in vacuolar Mg2+ sequestration that enables plant and yeast cells to cope with high levels of external Mg2+. We show that the yeast genome (as well as other fungal genomes) harbour a single ACDP homologue, referred to as MAM3, that functions specifically in vacuolar Mg2+ accumulation and is essential for tolerance to high Mg. In parallel, vacuolar ACDP homologues were identified from Arabidopsis and shown to complement the yeast mutant mam3Δ. An Arabidopsis mutant lacking one of the vacuolar ACDP homologues displayed hypersensitivity to high-Mg conditions and accumulated less Mg in the vacuole compared with the wild type. Taken together, our results suggest that conserved transporters mediate vacuolar Mg2+ sequestration in fungal and plant cells to maintain cellular Mg2+ homeostasis in response to fluctuating Mg2+ levels in the environment.
Asunto(s)
Proteínas de Arabidopsis , Saccharomyces cerevisiae , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Magnesio/metabolismo , Mutación , Células Vegetales/metabolismo , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismoRESUMEN
Magnesium (Mg2+ ) is an essential metal for plant growth; however, its over-accumulation in cells can be cytotoxic. The metal tolerance protein family (MTP) belongs to an ubiquitous family of cation diffusion facilitator (CDF) proteins that export divalent metal cations for metal homeostasis and tolerance in all organisms. We describe here the identification of MTP10 to be critical for xylem Mg homeostasis in Arabidopsis under high Mg2+ conditions. The Arabidopsis plant contains 12 MTP genes, and only knockout of MTP10 decreased the tolerance of high-Mg stress. The functional complementation assays in a Mg2+ -uptake-deficient bacterial strain MM281 confirmed that MTP10 conducted Mg2+ transport. MTP10 is localized to the plasma membrane of parenchyma cells around the xylem. Reciprocal grafting analysis further demonstrated that MTP10 functions in the shoot to determine the shoot growth phenotypes under high Mg2+ conditions. Moreover, compared to the wild type, the mtp10 mutant accumulated more Mg2+ in xylem sap under high-Mg stress. This study reveals that MTP10 facilitates Mg2+ diffusion from the xylem to shoots and thus determines Mg homeostasis in shoot vascular tissues during high-Mg stress.
Asunto(s)
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Regulación de la Expresión Génica de las Plantas , Magnesio/metabolismo , Raíces de Plantas/metabolismo , Brotes de la Planta/metabolismo , Xilema/metabolismoRESUMEN
The remodeling of root architecture is regarded as a major development to improve the plant's adaptivity to phosphate (Pi)-deficient conditions. The WRKY transcription factors family has been reported to regulate the Pi-deficiency-induced systemic responses by affecting Pi absorption or transportation. Whether these transcription factors act as a regulator to mediate the Pi-deficiency-induced remodeling of root architecture, a typical local response, is still unclear. Here, we identified an Arabidopsis transcription factor, WRKY33, that acted as a negative regulator to mediate the Pi-deficiency-induced remodeling of root architecture. The disruption of WRKY33 in wrky33-2 mutant increased the plant's low Pi sensitivity by further inhibiting the primary root growth and promoting the formation of root hair. Furthermore, we revealed that WRKY33 negatively regulated the remodeling of root architecture by controlling the transcriptional expression of ALMT1 under Pi-deficient conditions, which further mediated the Fe3+ accumulation in root tips to inhibit the root growth. In conclusion, this study demonstrates a previously unrecognized signaling crosstalk between WRKY33 and the ALMT1-mediated malate transport system to regulate the Pi deficiency responses.
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Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Hierro/metabolismo , Meristema/metabolismo , Transportadores de Anión Orgánico/metabolismo , Fosfatos/deficiencia , Factores de Transcripción/metabolismo , Arabidopsis/genética , Arabidopsis/fisiología , Proteínas de Arabidopsis/genética , Regulación de la Expresión Génica de las Plantas , Homeostasis , Meristema/genética , Meristema/fisiología , Transportadores de Anión Orgánico/genética , Raíces de Plantas/genética , Raíces de Plantas/metabolismo , Raíces de Plantas/fisiologíaRESUMEN
Nitrate (NO3- ) is a source of plant nutrients and osmolytes, but its delivery machineries under osmotic and low-nutrient stress remain largely unknown. Here, we report that AtICln, an Arabidopsis homolog of the nucleotide-sensitive chloride-conductance regulatory protein family (ICln), is involved in response to osmotic and low-NO3- stress. The gene AtICln, encoding plasma membrane-anchored proteins, was upregulated by various osmotic stresses, and its disruption impaired plant tolerance to osmotic stress. Compared with the wild type, the aticln mutant retained lower anions, particularly NO3- , and its growth retardation was not rescued by NO3- supply under osmotic stress. Interestingly, this mutant also displayed growth defects under low-NO3 stress, which were accompanied by decreases in NO3- accumulation, suggesting that AtICln may facilitate the NO3- accumulation under NO3- deficiency. Moreover, the low-NO3- hypersensitive phenotype of aticln mutant was overridden by the overexpression of NRT1.1, an important NO3- transporter in Arabidopsis low-NO3- responses. Further genetic analysis in the plants with altered activity of AtICln and NRT1.1 indicated that AtICln and NRT1.1 play a compensatory role in maintaining NO3- homeostasis under low-NO3- environments. These results suggest that AtICln is involved in cellular NO3- accumulation and thus determines osmotic adjustment and low-NO3- tolerance in plants.
Asunto(s)
Adaptación Fisiológica , Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Nitratos/metabolismo , Ósmosis , Homología de Secuencia de Aminoácido , Proteínas de Transporte de Anión/metabolismo , Membrana Celular/metabolismo , Cloruros/metabolismo , Prueba de Complementación Genética , Mutación/genética , Concentración Osmolar , Presión Osmótica , Fenotipo , Proteínas de Plantas/metabolismo , Fracciones Subcelulares/metabolismoRESUMEN
Type 2C protein phosphatases (PP2Cs) are the largest protein phosphatase family. PP2Cs dephosphorylate substrates for signaling in Arabidopsis, but the functions of most PP2Cs remain unknown. Here, we characterized PP2C49 (AT3G62260, a Group G PP2C), which regulates Na+ distribution under salt stress and is localized to the cytoplasm and nucleus. PP2C49 was highly expressed in root vascular tissues and its disruption enhanced plant tolerance to salt stress. Compared with wild type, the pp2c49 mutant contained more Na+ in roots but less Na+ in shoots and xylem sap, suggesting that PP2C49 regulates shoot Na+ extrusion. Reciprocal grafting revealed a root-based mechanism underlying the salt tolerance of pp2c49. Systemic Na+ distribution largely depends on AtHKT1;1 and loss of function of AtHKT1;1 in the pp2c49 background overrode the salt tolerance of pp2c49, resulting in salt sensitivity. Furthermore, compared with plants overexpressing PP2C49 in the wild-type background, plants overexpressing PP2C49 in the athtk1;1 mutant background were sensitive to salt, like the athtk1;1 mutants. Moreover, protein-protein interaction and two-voltage clamping assays demonstrated that PP2C49 physically interacts with AtHKT1;1 and inhibits the Na+ permeability of AtHKT1;1. This study reveals that PP2C49 negatively regulates AtHKT1;1 activity and thus determines systemic Na+ allocation during salt stress.
Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/enzimología , Arabidopsis/fisiología , Proteínas de Transporte de Catión/antagonistas & inhibidores , Proteína Fosfatasa 2C/metabolismo , Tolerancia a la Sal/fisiología , Simportadores/antagonistas & inhibidores , Ácido Abscísico/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/antagonistas & inhibidores , Proteínas de Arabidopsis/genética , Proteínas de Transporte de Catión/metabolismo , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Mutación/genética , Fenotipo , Raíces de Plantas/efectos de los fármacos , Raíces de Plantas/metabolismo , Brotes de la Planta/efectos de los fármacos , Brotes de la Planta/metabolismo , Unión Proteica/efectos de los fármacos , Proteína Fosfatasa 2C/genética , Transducción de Señal/efectos de los fármacos , Sodio/metabolismo , Cloruro de Sodio/farmacología , Fracciones Subcelulares/efectos de los fármacos , Fracciones Subcelulares/metabolismo , Simportadores/metabolismo , Xilema/metabolismoRESUMEN
Chlorophyll (Chl) is essential for photosynthetic reactions and chloroplast development. While the enzymatic pathway for Chl biosynthesis is well established, the regulatory mechanism underlying the homeostasis of Chl levels remains largely unknown. In this study, we identified CBD1 (Chlorophyll Biosynthetic Defect1), which functions in the regulation of chlorophyll biosynthesis. The CBD1 gene was expressed specifically in green tissues and its protein product was embedded in the thylakoid membrane. Furthermore, CBD1 was precisely co-expressed and functionally correlated with GUN5 (Genome Uncoupled 5). Analysis of chlorophyll metabolic intermediates indicated that cbd1 and cbd1gun5 mutants over-accumulated magnesium protoporphyrin IX (Mg-Proto IX). In addition, the cbd1 mutant thylakoid contained less Mg than the wild type not only as a result of lower Chl content, but also implicating CBD1 in Mg transport. This was supported by the finding that CBD1 complemented a Mg2+ uptake-deficient Salmonella strain under low Mg conditions. Taken together, these results indicate that CBD1 functions synergistically with CHLH/GUN5 in Mg-Proto IX processing, and may serve as a Mg-transport protein to maintain Mg homeostasis in the chloroplast.
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Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Clorofila/biosíntesis , Liasas/metabolismo , Proteínas de las Membranas de los Tilacoides/metabolismo , Proteína 9 Asociada a CRISPR , Sistemas CRISPR-Cas , Cloroplastos/metabolismo , Cloroplastos/ultraestructura , Edición Génica , Técnicas de Silenciamiento del Gen , Homeostasis , Magnesio/metabolismo , Microscopía Electrónica de Transmisión , Tilacoides/metabolismoRESUMEN
Plant elicitor peptides (Peps) are damage/danger-associated molecular patterns (DAMPs) that are derived from precursor proteins PROPEPs and perceived by a pair of leucine-rich repeat receptor-like kinases (LRR-RLKs), PEPR1 and PEPR2, to enhance innate immunity and to inhibit root growth in Arabidopsis thaliana. In this study, we show that Arabidopsis Pep1 inhibits the root growth by interfering with pH signaling, as acidic condition increased, but neutral and alkaline conditions decreased the Pep1 effect on inhibiting the root growth. The perception of Pep1 to PEPRs activated the plasma membrane-localized H+-ATPases (PM H+-ATPases) -the pump proton in plant cell-to extrude the protons into apoplast, and induced an overly acidic environment in apoplastic space, which further promoted the cell swelling in root apex and inhibited root growth. Furthermore, we revealed that pump proton AUTOINHIBITED H+-ATPase 2 (AHA2) physically interacted with PEPR2 and served downstream of the Pep1-PEPRs signaling pathway to regulate Pep1-induced protons extrusion and root growth inhibition. In conclusion, this study demonstrates a previously unrecognized signaling crosstalk between Pep1 and pH signaling to regulate root growth.
Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/crecimiento & desarrollo , Proteínas Serina-Treonina Quinasas/metabolismo , ATPasas de Translocación de Protón/metabolismo , Transactivadores/metabolismo , Alarminas/metabolismo , Arabidopsis/metabolismo , Membrana Celular/metabolismo , Regulación del Desarrollo de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Concentración de Iones de Hidrógeno , Raíces de Plantas/crecimiento & desarrollo , Raíces de Plantas/metabolismo , Transducción de SeñalRESUMEN
Plant elicitor peptides (Peps) are damage/danger-associated molecular patterns (DAMPs) that are perceived by a pair of receptor-like kinases, PEPR1 and PEPR2, to enhance innate immunity and induce the growth inhibition of root in Arabidopsis thaliana. In this study, we show that PEPR1 and PEPR2 function vitally in roots to regulate the root immune responses when treating the roots with bacterial pathogen Pst DC3000. PEPR2, rather than PEPR1, played a predominant role in the perception of Pep1 in the roots and further triggered a strong ROS accumulation-the substance acts as an antimicrobial agent or as a secondary messenger in plant cells. Consistently, seedlings mutating two major ROS-generating enzyme genes, respiratory burst oxidase homologs D and F (RBOHD and RBOHF), abolished the root ROS accumulation and impaired the growth inhibition of the roots induced by Pep1. Furthermore, we revealed that botrytis-induced kinase 1 (BIK1) physically interacted with PEPRs and RBOHD/F, respectively, and served downstream of the Pep1-PEPRs signaling pathway to regulate Pep1-induced ROS production and root growth inhibition. In conclusion, this study demonstrates a previously unrecognized signaling crosstalk between Pep1 and ROS signaling to regulate root immune response and root growth.
Asunto(s)
Alarminas/metabolismo , Proteínas de Arabidopsis/metabolismo , Arabidopsis/crecimiento & desarrollo , Fragmentos de Péptidos/farmacología , Inmunidad de la Planta/efectos de los fármacos , Raíces de Plantas/crecimiento & desarrollo , Especies Reactivas de Oxígeno/metabolismo , Alarminas/genética , Arabidopsis/efectos de los fármacos , Arabidopsis/inmunología , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Raíces de Plantas/efectos de los fármacos , Raíces de Plantas/inmunología , Raíces de Plantas/metabolismo , Proteínas Serina-Treonina Quinasas/genética , Proteínas Serina-Treonina Quinasas/metabolismo , Receptores de Superficie Celular/genética , Receptores de Superficie Celular/metabolismoRESUMEN
The Arabidopsis genome comprises eighty genes encoding BTB (broad-complex, tramtrack, and bric-a-brac) family proteins that are characterized with the BTB domain and that potentially serve as substrate adaptors for cullin-based E3-ligases. In addition to the BTB domain, most BTB proteins also contain various other interaction motifs that probably act as target recognition elements. Here, we report three members of the BTB-A2 subfamily that distinctly only contain the BTB domain, BTB-A2.1, BTB-A2.2, and BTB-A2.3, that negatively regulates abscisic acid (ABA) signaling in Arabidopsis. BTB-A2.1, BTB-A2.2, and BTB-A2.3 encoded cytoplasm- and nucleus-localized proteins and displayed highly overlapping expression patterns in Arabidopsis tissues. Disruption of these three genes, but not single or double mutants, resulted in a decrease in ABA-induced inhibition of seed germination. Further analyses demonstrated the expression levels of these three genes were up-regulated by ABA, and their mutation increased ABA signalling. Importantly, protein-protein interaction assays showed that these three BTB-A2 proteins physically interacted with SnRK2.3. Moreover, biochemical and genetic assays indicated that BTB-A2.1, BTB-A2.2, and BTB-A2.3 decreased the stability of SnRK2.3 and attenuated the SnRK2.3 responsible for the ABA hypersensitive phenotype of seed germination. This report thus reveals that BTB-A2s serve as negative regulators for balancing the intensity of ABA signaling during seed germination.
Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/fisiología , Germinación , Proteínas Serina-Treonina Quinasas/metabolismo , Semillas/fisiología , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Proteínas de Arabidopsis/genética , Expresión Génica , Germinación/genética , Fenotipo , Unión Proteica , Estabilidad Proteica , Transporte de Proteínas , Transducción de SeñalRESUMEN
Ethylene is a gaseous phytohormone that is perceived by two-component histidine kinase-type receptors. Recent studies identified choline transporter-like 1 (CTL1) essential for Arabidopsis growth and development, including apical hook development in the etiolated seedlings. Here, we report that CTL1 contributes to apical hook development by enhancing ethylene response. The expression of CTL1 was highly correlated with the intensity of ethylene response and was enriched in the apical hook, cotyledon tip and hypocotyl. Genetic analysis showed that the dark-grown ctl1 mutant displayed a defect in ethylene-induced apical hook development as compared with the wild type. Accordingly, the expression of ethylene signaling reporter EBS::GUS in ctl1 mutant was greatly reduced in leaves, apical hook, hypocotyl and root, suggesting that the disruption of CTL1 impairs the ethylene signaling. Furthermore, protein-protein interaction assays demonstrated that CTL1 may interact with ethylene receptors, including ETR1, ETR2, ERS1, ERS2. Importantly, the abundance of CTL1 was diminished when ETR1 was disrupted upon ethylene response. Taken together, our results suggest that CTL1 functions as a positive regulator in ethylene signaling which in turn contributes to apical hook development of etiolated plant seedlings.
Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/crecimiento & desarrollo , Proteínas de Transporte de Membrana/metabolismo , Plantones/crecimiento & desarrollo , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Etilenos/metabolismo , Regulación de la Expresión Génica de las Plantas , Proteínas de Transporte de Membrana/genética , Reguladores del Crecimiento de las Plantas/metabolismo , Plantones/genética , Plantones/metabolismo , Regulación hacia ArribaRESUMEN
Membrane transport processes are indispensable for many aspects of plant physiology including mineral nutrition, solute storage, cell metabolism, cell signaling, osmoregulation, cell growth, and stress responses. Completion of genome sequencing in diverse plant species and the development of multiple genomic tools have marked a new era in understanding plant membrane transport at the mechanistic level. Genes coding for a galaxy of pumps, channels, and carriers that facilitate various membrane transport processes have been identified while multiple approaches are developed to dissect the physiological roles as well as to define the transport capacities of these transport systems. Furthermore, signaling networks dictating the membrane transport processes are established to fully understand the regulatory mechanisms. Here, we review recent research progress in the discovery and characterization of the components in plant membrane transport that take advantage of plant genomic resources and other experimental tools. We also provide our perspectives for future studies in the field.
Asunto(s)
Membrana Celular/metabolismo , Proteínas de Plantas/metabolismo , Plantas/metabolismo , Genética Inversa/métodos , Transporte Biológico , Membrana Celular/genética , Genoma de Planta , Genómica/métodos , Familia de Multigenes , Proteínas de Plantas/genética , Plantas/genética , Transducción de SeñalRESUMEN
Plants cope with aluminum (Al) toxicity by secreting organic acids (OAs) into the apoplastic space, which is driven by proton (H+) pumps. Here, we show that mutation of vacuolar H+-translocating adenosine triphosphatase (H+-ATPase) subunit a2 (VHA-a2) and VHA-a3 of the vacuolar H+-ATPase enhances Al resistance in Arabidopsis (Arabidopsis thaliana). vha-a2 vha-a3 mutant plants displayed less Al sensitivity with less Al accumulation in roots compared to wild-type plants when grown under excessive Al3+ Interestingly, in response to Al3+ exposure, plants showed decreased vacuolar H+ pump activity and reduced expression of VHA-a2 and VHA-a3, which were accompanied by increased plasma membrane H+ pump (PM H+-ATPase) activity. Genetic analysis of plants with altered PM H+-ATPase activity established a correlation between Al-induced increase in PM H+-ATPase activity and enhanced Al resistance in vha-a2 vha-a3 plants. We determined that external OAs, such as malate and citrate whose secretion is driven by PM H+-ATPase, increased with PM H+-ATPase activity upon Al stress. On the other hand, elevated secretion of malate and citrate in vha-a2 vha-a3 root exudates appeared to be independent of OAs metabolism and tolerance of phosphate starvation but was likely related to impaired vacuolar sequestration. These results suggest that coordination of vacuolar H+-ATPase and PM H+-ATPase dictates the distribution of OAs into either the vacuolar lumen or the apoplastic space that, in turn, determines Al tolerance capacity in plants.