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1.
BMC Med Educ ; 24(1): 352, 2024 Mar 29.
Artículo en Inglés | MEDLINE | ID: mdl-38553736

RESUMEN

Blended Intensive Programmes (BIP's) represent a valuable tool for gathering knowledge and summarising the latest trends in medicine and dentistry. Blended education has been found, even before the COVID-19 pandemic, to increase the level of education and stimulate effective learning for postgraduate healthcare professionals. Interprofessional education is critical for preparing students to enter the health workforce, where teamwork and collaboration are important competencies. This article outlines the key points of the Blended Intensive Programme's implementation in dental education organised by Wroclaw Medical University in Poland. BIP involved professors from 12 universities or research institutions from Europe and South America and 28 participants from 8 countries. The course was taught remotely and in person. In addition, it included a visit to the university and practical classes with artificial simulation and practice in dentistry. A structured questionnaire enabled measuring the evaluation of students' perception of the COVID-19 education before and after the pandemic. The European Region Action Scheme for the Mobility of University Students (ERASMUS) was fundamental to carrying out the BIP with the participation of several countries, allowing the exchange of knowledge, assessing the impact of the pandemic on dental universities, and strengthening international collaborations and the future project of research, education and clinical assistance. We conclude that hybrid teaching programmes broaden the learning spectrum in dental studies by allowing transnational and interdisciplinary approaches that make students aware of the importance of their work within the framework of the general health approach, as this differs from country to country.


Asunto(s)
Aprendizaje , Pandemias , Humanos , Curriculum , Estudiantes , Educación en Odontología
2.
Plant J ; 106(3): 601-615, 2021 05.
Artículo en Inglés | MEDLINE | ID: mdl-33544927

RESUMEN

Pattern-triggered immunity (PTI) is activated in plants upon recognition by pattern recognition receptors (PRRs) of damage- and microbe-associated molecular patterns (DAMPs and MAMPs) derived from plants or microorganisms, respectively. To understand better the plant mechanisms involved in the perception of carbohydrate-based structures recognized as DAMPs/MAMPs, we have studied the ability of mixed-linked ß-1,3/1,4-glucans (MLGs), present in some plant and microbial cell walls, to trigger immune responses and disease resistance in plants. A range of MLG structures were tested for their capacity to induce PTI hallmarks, such as cytoplasmic Ca2+ elevations, reactive oxygen species production, phosphorylation of mitogen-activated protein kinases and gene transcriptional reprogramming. These analyses revealed that MLG oligosaccharides are perceived by Arabidopsis thaliana and identified a trisaccharide, ß-d-cellobiosyl-(1,3)-ß-d-glucose (MLG43), as the smallest MLG structure triggering strong PTI responses. These MLG43-mediated PTI responses are partially dependent on LysM PRRs CERK1, LYK4 and LYK5, as they were weaker in cerk1 and lyk4 lyk5 mutants than in wild-type plants. Cross-elicitation experiments between MLG43 and the carbohydrate MAMP chitohexaose [ß-1,4-d-(GlcNAc)6 ], which is also perceived by these LysM PRRs, indicated that the mechanism of MLG43 recognition could differ from that of chitohexaose, which is fully impaired in cerk1 and lyk4 lyk5 plants. MLG43 treatment confers enhanced disease resistance in A. thaliana to the oomycete Hyaloperonospora arabidopsidis and in tomato and pepper to different bacterial and fungal pathogens. Our data support the classification of MLGs as a group of carbohydrate-based molecular patterns that are perceived by plants and trigger immune responses and disease resistance.


Asunto(s)
Pared Celular/metabolismo , Resistencia a la Enfermedad , Inmunidad de la Planta , beta-Glucanos/metabolismo , Arabidopsis/inmunología , Arabidopsis/metabolismo , Calcio/metabolismo , Capsicum/inmunología , Capsicum/metabolismo , Solanum lycopersicum/inmunología , Solanum lycopersicum/metabolismo , Oomicetos/inmunología , Enfermedades de las Plantas/inmunología , Enfermedades de las Plantas/microbiología , Trisacáridos
3.
J Exp Bot ; 73(1): 339-350, 2022 01 05.
Artículo en Inglés | MEDLINE | ID: mdl-34463334

RESUMEN

Zinc is an essential nutrient at low concentrations, but toxic at slightly higher ones. It has been proposed that hyperaccumulator plants may use the excess zinc to fend off pathogens and herbivores. However, there is little evidence of a similar response in other plants. Here we show that Arabidopsis thaliana leaves inoculated with the necrotrophic fungus Plectosphaerella cucumerina BMM (PcBMM) accumulate zinc and manganese at the infection site. Zinc accumulation did not occur in a double mutant in the zinc transporters HEAVY METAL ATPASE2 and HEAVY METAL ATPASE4 (HMA2 and HMA4), which has reduced zinc translocation from roots to shoots. Consistent with a role in plant immunity, expression of HMA2 and HMA4 was up-regulated upon PcBMM inoculation, and hma2hma4 mutants were more susceptible to PcBMM infection. This phenotype was rescued upon zinc supplementation. The increased susceptibility to PcBMM infection was not due to the diminished expression of genes involved in the salicylic acid, ethylene, or jasmonate pathways since they were constitutively up-regulated in hma2hma4 plants. Our data indicate a role of zinc in resistance to PcBMM in plants containing ordinary levels of zinc. This layer of immunity runs in parallel to the already characterized defence pathways, and its removal has a direct effect on resistance to pathogens.


Asunto(s)
Proteínas de Arabidopsis , Arabidopsis , Ascomicetos , Adenosina Trifosfatasas/genética , Adenosina Trifosfatasas/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Regulación de la Expresión Génica de las Plantas , Zinc/metabolismo
4.
Plant J ; 93(1): 34-49, 2018 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-29083116

RESUMEN

Fungal cell walls, which are essential for environmental adaptation and host colonization by the fungus, have been evolutionarily selected by plants and animals as a source of microbe-associated molecular patterns (MAMPs) that, upon recognition by host pattern recognition receptors (PRRs), trigger immune responses conferring disease resistance. Chito-oligosaccharides [ß-1,4-N-acetylglucosamine oligomers, (GlcNAc)n ] are the only glycosidic structures from fungal walls that have been well-demonstrated to function as MAMPs in plants. Perception of (GlcNAc)4-8 by Arabidopsis involves CERK1, LYK4 and LYK5, three of the eight members of the LysM PRR family. We found that a glucan-enriched wall fraction from the pathogenic fungus Plectosphaerella cucumerina which was devoid of GlcNAc activated immune responses in Arabidopsis wild-type plants but not in the cerk1 mutant. Using this differential response, we identified the non-branched 1,3-ß-d-(Glc) hexasaccharide as a major fungal MAMP. Recognition of 1,3-ß-d-(Glc)6 was impaired in cerk1 but not in mutants defective in either each of the LysM PRR family members or in the PRR-co-receptor BAK1. Transcriptomic analyses of Arabidopsis plants treated with 1,3-ß-d-(Glc)6 further demonstrated that this fungal MAMP triggers the expression of immunity-associated genes. In silico docking analyses with molecular mechanics and solvation energy calculations corroborated that CERK1 can bind 1,3-ß-d-(Glc)6 at effective concentrations similar to those of (GlcNAc)4 . These data support that plants, like animals, have selected as MAMPs the linear 1,3-ß-d-glucans present in the walls of fungi and oomycetes. Our data also suggest that CERK1 functions as an immune co-receptor for linear 1,3-ß-d-glucans in a similar way to its proposed function in the recognition of fungal chito-oligosaccharides and bacterial peptidoglycan MAMPs.


Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/inmunología , Ascomicetos/fisiología , Enfermedades de las Plantas/inmunología , Inmunidad de la Planta/efectos de los fármacos , Proteínas Serina-Treonina Quinasas/metabolismo , beta-Glucanos/farmacología , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Pared Celular/metabolismo , Oligosacáridos/farmacología , Enfermedades de las Plantas/microbiología , Proteínas Serina-Treonina Quinasas/genética , Receptores de Reconocimiento de Patrones/metabolismo
5.
Mol Plant Microbe Interact ; 32(4): 464-478, 2019 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-30387369

RESUMEN

Genetic ablation of the ß subunit of the heterotrimeric G protein complex in agb1-2 confers defective activation of microbe-associated molecular pattern (MAMP)-triggered immunity, resulting in agb1-2 enhanced susceptibility to pathogens like the fungus Plectosphaerella cucumerina BMM. A mutant screen for suppressors of agb1-2 susceptibility (sgb) to P. cucumerina BMM identified sgb10, a new null allele (mkp1-2) of the mitogen-activated protein kinase phosphatase 1 (MKP1). The enhanced susceptibility of agb1-2 to the bacterium Pseudomonas syringae pv. tomato DC3000 and the oomycete Hyaloperonospora arabidopsidis is also abrogated by mkp1-2. MKP1 negatively balances production of reactive oxygen species (ROS) triggered by MAMPs, since ROS levels are enhanced in mkp1. The expression of RBOHD, encoding a NADPH oxidase-producing ROS, is upregulated in mkp1 upon MAMP treatment or pathogen infection. Moreover, MKP1 negatively regulates RBOHD activity, because ROS levels upon MAMP treatment are increased in mkp1 plants constitutively overexpressing RBOHD (35S::RBOHD mkp1). A significant reprograming of mkp1 metabolic profile occurs with more than 170 metabolites, including antimicrobial compounds, showing differential accumulation in comparison with wild-type plants. These results suggest that MKP1 functions downstream of the heterotrimeric G protein during MAMP-triggered immunity, directly regulating the activity of RBOHD and ROS production as well as other immune responses.


Asunto(s)
Proteínas de Arabidopsis , Arabidopsis , Resistencia a la Enfermedad , Regulación de la Expresión Génica de las Plantas , Proteínas Tirosina Fosfatasas , Arabidopsis/inmunología , Arabidopsis/microbiología , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Ascomicetos/fisiología , Resistencia a la Enfermedad/genética , Regulación de la Expresión Génica de las Plantas/genética , Proteínas Tirosina Fosfatasas/genética , Proteínas Tirosina Fosfatasas/metabolismo , Pseudomonas syringae/fisiología , Especies Reactivas de Oxígeno/metabolismo
6.
Plant J ; 92(3): 386-399, 2017 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-28792629

RESUMEN

Arabidopsis heterotrimeric G-protein complex modulates pathogen-associated molecular pattern-triggered immunity (PTI) and disease resistance responses to different types of pathogens. It also plays a role in plant cell wall integrity as mutants impaired in the Gß- (agb1-2) or Gγ-subunits have an altered wall composition compared with wild-type plants. Here we performed a mutant screen to identify suppressors of agb1-2 (sgb) that restore susceptibility to pathogens to wild-type levels. Out of the four sgb mutants (sgb10-sgb13) identified, sgb11 is a new mutant allele of ESKIMO1 (ESK1), which encodes a plant-specific polysaccharide O-acetyltransferase involved in xylan acetylation. Null alleles (sgb11/esk1-7) of ESK1 restore to wild-type levels the enhanced susceptibility of agb1-2 to the necrotrophic fungus Plectosphaerella cucumerina BMM (PcBMM), but not to the bacterium Pseudomonas syringae pv. tomato DC3000 or to the oomycete Hyaloperonospora arabidopsidis. The enhanced resistance to PcBMM of the agb1-2 esk1-7 double mutant was not the result of the re-activation of deficient PTI responses in agb1-2. Alteration of cell wall xylan acetylation caused by ESK1 impairment was accompanied by an enhanced accumulation of abscisic acid, the constitutive expression of genes encoding antibiotic peptides and enzymes involved in the biosynthesis of tryptophan-derived metabolites, and the accumulation of disease resistance-related secondary metabolites and different osmolites. These esk1-mediated responses counterbalance the defective PTI and PcBMM susceptibility of agb1-2 plants, and explain the enhanced drought resistance of esk1 plants. These results suggest that a deficient PTI-mediated resistance is partially compensated by the activation of specific cell-wall-triggered immune responses.


Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/genética , Proteínas de Unión al GTP Heterotriméricas/metabolismo , Enfermedades de las Plantas/inmunología , Inmunidad de la Planta/genética , Xilanos/metabolismo , Ácido Abscísico/metabolismo , Acetilación , Acetiltransferasas , Arabidopsis/inmunología , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Ascomicetos/fisiología , Pared Celular/metabolismo , Subunidades beta de la Proteína de Unión al GTP/genética , Subunidades beta de la Proteína de Unión al GTP/metabolismo , Regulación de la Expresión Génica de las Plantas , Proteínas de Unión al GTP Heterotriméricas/genética , Proteínas de la Membrana , Modelos Biológicos , Mutación , Enfermedades de las Plantas/microbiología , Reguladores del Crecimiento de las Plantas/metabolismo , Pseudomonas syringae/fisiología , Plantones/genética , Plantones/inmunología , Plantones/metabolismo
7.
New Phytol ; 218(2): 661-680, 2018 04.
Artículo en Inglés | MEDLINE | ID: mdl-29451312

RESUMEN

Mitogen-activated protein kinases (MAPKs) cascades play essential roles in plants by transducing developmental cues and environmental signals into cellular responses. Among the latter are microbe-associated molecular patterns perceived by pattern recognition receptors (PRRs), which trigger immunity. We found that YODA (YDA) - a MAPK kinase kinase regulating several Arabidopsis developmental processes, like stomatal patterning - also modulates immune responses. Resistance to pathogens is compromised in yda alleles, whereas plants expressing the constitutively active YDA (CA-YDA) protein show broad-spectrum resistance to fungi, bacteria, and oomycetes with different colonization modes. YDA functions in the same pathway as ERECTA (ER) Receptor-Like Kinase, regulating both immunity and stomatal patterning. ER-YDA-mediated immune responses act in parallel to canonical disease resistance pathways regulated by phytohormones and PRRs. CA-YDA plants exhibit altered cell-wall integrity and constitutively express defense-associated genes, including some encoding putative small secreted peptides and PRRs whose impairment resulted in enhanced susceptibility phenotypes. CA-YDA plants show strong reprogramming of their phosphoproteome, which contains protein targets distinct from described MAPKs substrates. Our results suggest that, in addition to stomata development, the ER-YDA pathway regulates an immune surveillance system conferring broad-spectrum disease resistance that is distinct from the canonical pathways mediated by described PRRs and defense hormones.


Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/enzimología , Arabidopsis/inmunología , Resistencia a la Enfermedad , Quinasas Quinasa Quinasa PAM/metabolismo , Enfermedades de las Plantas/inmunología , Enfermedades de las Plantas/microbiología , Inmunidad de la Planta , Tipificación del Cuerpo , Pared Celular/metabolismo , Flagelina/farmacología , Hongos/fisiología , Regulación de la Expresión Génica de las Plantas , Modelos Biológicos , Mutación/genética , Moléculas de Patrón Molecular Asociado a Patógenos/metabolismo , Estomas de Plantas/crecimiento & desarrollo , Transducción de Señal , Regulación hacia Arriba/genética
8.
Plant Physiol ; 165(2): 791-809, 2014 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-24755512

RESUMEN

Systemic acquired resistance (SAR) is an inducible immune response that depends on ENHANCED DISEASE SUSCEPTIBILITY1 (EDS1). Here, we show that Arabidopsis (Arabidopsis thaliana) EDS1 is required for both SAR signal generation in primary infected leaves and SAR signal perception in systemic uninfected tissues. In contrast to SAR signal generation, local resistance remains intact in eds1 mutant plants in response to Pseudomonas syringae delivering the effector protein AvrRpm1. We utilized the SAR-specific phenotype of the eds1 mutant to identify new SAR regulatory proteins in plants conditionally expressing AvrRpm1. Comparative proteomic analysis of apoplast-enriched extracts from AvrRpm1-expressing wild-type and eds1 mutant plants led to the identification of 12 APOPLASTIC, EDS1-DEPENDENT (AED) proteins. The genes encoding AED1, a predicted aspartyl protease, and another AED, LEGUME LECTIN-LIKE PROTEIN1 (LLP1), were induced locally and systemically during SAR signaling and locally by salicylic acid (SA) or its functional analog, benzo 1,2,3-thiadiazole-7-carbothioic acid S-methyl ester. Because conditional overaccumulation of AED1-hemagglutinin inhibited SA-induced resistance and SAR but not local resistance, the data suggest that AED1 is part of a homeostatic feedback mechanism regulating systemic immunity. In llp1 mutant plants, SAR was compromised, whereas the local resistance that is normally associated with EDS1 and SA as well as responses to exogenous SA appeared largely unaffected. Together, these data indicate that LLP1 promotes systemic rather than local immunity, possibly in parallel with SA. Our analysis reveals new positive and negative components of SAR and reinforces the notion that SAR represents a distinct phase of plant immunity beyond local resistance.

9.
Hortic Res ; 11(1): uhad248, 2024 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-38239809

RESUMEN

Biotic and abiotic stresses can severely limit crop productivity. In response to drought, plants close stomata to prevent water loss. Furthermore, stomata are the main entry point for several pathogens. Therefore, the development of natural products to control stomata closure can be considered a sustainable strategy to cope with stresses in agriculture. Plants respond to different stresses by releasing volatile organic compounds. Green leaf volatiles, which are commonly produced across different plant species after tissue damage, comprise an important group within volatile organic compounds. Among them, (Z)-3-hexenyl butyrate (HB) was described as a natural inducer of stomatal closure, playing an important role in stomatal immunity, although its mechanism of action is still unknown. Through different genetic, pharmacological, and biochemical approaches, we here uncover that HB perception initiates various defence signalling events, such as activation of Ca2+ permeable channels, mitogen-activated protein kinases, and production of Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase-mediated reactive oxygen species. Furthermore, HB-mediated stomata closure was found to be independent of abscisic acid biosynthesis and signalling. Additionally, exogenous treatments with HB alleviate water stress and improve fruit productivity in tomato plants. The efficacy of HB was also tested under open field conditions, leading to enhanced resistance against Phytophthora spp. and Pseudomonas syringae infection in potato and tomato plants, respectively. Taken together, our results provide insights into the HB signalling transduction pathway, confirming its role in stomatal closure and plant immune system activation, and propose HB as a new phytoprotectant for the sustainable control of biotic and abiotic stresses in agriculture.

10.
Mol Plant ; 17(5): 699-724, 2024 May 06.
Artículo en Inglés | MEDLINE | ID: mdl-38594902

RESUMEN

Beyond their function as structural barriers, plant cell walls are essential elements for the adaptation of plants to environmental conditions. Cell walls are dynamic structures whose composition and integrity can be altered in response to environmental challenges and developmental cues. These wall changes are perceived by plant sensors/receptors to trigger adaptative responses during development and upon stress perception. Plant cell wall damage caused by pathogen infection, wounding, or other stresses leads to the release of wall molecules, such as carbohydrates (glycans), that function as damage-associated molecular patterns (DAMPs). DAMPs are perceived by the extracellular ectodomains (ECDs) of pattern recognition receptors (PRRs) to activate pattern-triggered immunity (PTI) and disease resistance. Similarly, glycans released from the walls and extracellular layers of microorganisms interacting with plants are recognized as microbe-associated molecular patterns (MAMPs) by specific ECD-PRRs triggering PTI responses. The number of oligosaccharides DAMPs/MAMPs identified that are perceived by plants has increased in recent years. However, the structural mechanisms underlying glycan recognition by plant PRRs remain limited. Currently, this knowledge is mainly focused on receptors of the LysM-PRR family, which are involved in the perception of various molecules, such as chitooligosaccharides from fungi and lipo-chitooligosaccharides (i.e., Nod/MYC factors from bacteria and mycorrhiza, respectively) that trigger differential physiological responses. Nevertheless, additional families of plant PRRs have recently been implicated in oligosaccharide/polysaccharide recognition. These include receptor kinases (RKs) with leucine-rich repeat and Malectin domains in their ECDs (LRR-MAL RKs), Catharanthus roseus RECEPTOR-LIKE KINASE 1-LIKE group (CrRLK1L) with Malectin-like domains in their ECDs, as well as wall-associated kinases, lectin-RKs, and LRR-extensins. The characterization of structural basis of glycans recognition by these new plant receptors will shed light on their similarities with those of mammalians involved in glycan perception. The gained knowledge holds the potential to facilitate the development of sustainable, glycan-based crop protection solutions.


Asunto(s)
Pared Celular , Resistencia a la Enfermedad , Pared Celular/metabolismo , Enfermedades de las Plantas/microbiología , Enfermedades de las Plantas/inmunología , Receptores de Reconocimiento de Patrones/metabolismo , Plantas/metabolismo , Plantas/microbiología , Plantas/inmunología , Inmunidad de la Planta/fisiología
11.
Children (Basel) ; 11(3)2024 Mar 18.
Artículo en Inglés | MEDLINE | ID: mdl-38539393

RESUMEN

External apical root resorption in permanent teeth is a multifactorial process influenced by a variety of local and systemic factors. This report describes a case of multiple and severe apical root resorptions in a patient with Turner syndrome. The condition was discovered in a young female with Turner syndrome after 30 months of orthodontic treatment with fixed appliance. The purpose of this report is to present reports by other authors on the potential causes of the increased risk of tooth resorption in patients with Turner syndrome and to share insights derived from its course, highlighting the implications and lessons learned. Patients with Turner syndrome are not ideal candidates for orthodontic treatment. Prior to commencing orthodontic treatment, it is essential to carefully consider the potential benefits of the therapy compared to the risk associated with exacerbating root resorption. In the case of Turner syndrome patients, where there is an elevated risk of such complications, a thorough analysis should be conducted to determine whether the expected benefits of the treatment outweigh the potential hazards to the patient's dental health.

12.
EMBO J ; 28(21): 3428-38, 2009 Nov 04.
Artículo en Inglés | MEDLINE | ID: mdl-19763086

RESUMEN

In plant innate immunity, the surface-exposed leucine-rich repeat receptor kinases EFR and FLS2 mediate recognition of the bacterial pathogen-associated molecular patterns EF-Tu and flagellin, respectively. We identified the Arabidopsis stromal-derived factor-2 (SDF2) as being required for EFR function, and to a lesser extent FLS2 function. SDF2 resides in an endoplasmic reticulum (ER) protein complex with the Hsp40 ERdj3B and the Hsp70 BiP, which are components of the ER-quality control (ER-QC). Loss of SDF2 results in ER retention and degradation of EFR. The differential requirement for ER-QC components by EFR and FLS2 could be linked to N-glycosylation mediated by STT3a, a catalytic subunit of the oligosaccharyltransferase complex involved in co-translational N-glycosylation. Our results show that the plasma membrane EFR requires the ER complex SDF2-ERdj3B-BiP for its proper accumulation, and provide a demonstration of a physiological requirement for ER-QC in transmembrane receptor function in plants. They also provide an unexpected differential requirement for ER-QC and N-glycosylation components by two closely related receptors.


Asunto(s)
Proteínas de Arabidopsis/inmunología , Arabidopsis/inmunología , Retículo Endoplásmico/metabolismo , Enfermedades de las Plantas/inmunología , Receptores de Reconocimiento de Patrones/inmunología , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Proteínas Portadoras/metabolismo , Proteínas del Choque Térmico HSP40/metabolismo , Inmunidad Innata , Receptores de Reconocimiento de Patrones/genética , Receptores de Reconocimiento de Patrones/metabolismo
13.
Mol Syst Biol ; 7: 532, 2011 Sep 27.
Artículo en Inglés | MEDLINE | ID: mdl-21952135

RESUMEN

The heterotrimeric G-protein complex is minimally composed of Gα, Gß, and Gγ subunits. In the classic scenario, the G-protein complex is the nexus in signaling from the plasma membrane, where the heterotrimeric G-protein associates with heptahelical G-protein-coupled receptors (GPCRs), to cytoplasmic target proteins called effectors. Although a number of effectors are known in metazoans and fungi, none of these are predicted to exist in their canonical forms in plants. To identify ab initio plant G-protein effectors and scaffold proteins, we screened a set of proteins from the G-protein complex using two-hybrid complementation in yeast. After deep and exhaustive interrogation, we detected 544 interactions between 434 proteins, of which 68 highly interconnected proteins form the core G-protein interactome. Within this core, over half of the interactions comprising two-thirds of the nodes were retested and validated as genuine in planta. Co-expression analysis in combination with phenotyping of loss-of-function mutations in a set of core interactome genes revealed a novel role for G-proteins in regulating cell wall modification.


Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis , Pared Celular , Proteínas de Unión al GTP/metabolismo , Glicómica , Proteómica , Receptores Acoplados a Proteínas G/metabolismo , Transducción de Señal/genética , Arabidopsis/genética , Arabidopsis/crecimiento & desarrollo , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Membrana Celular/genética , Membrana Celular/metabolismo , Pared Celular/genética , Pared Celular/metabolismo , Bases de Datos Genéticas , Proteínas de Unión al GTP/genética , Regulación de la Expresión Génica de las Plantas , Redes Reguladoras de Genes , Prueba de Complementación Genética , Genotipo , Inmunoprecipitación , Morfogénesis/genética , Fenotipo , Mapeo de Interacción de Proteínas , Receptores Acoplados a Proteínas G/genética , Técnicas del Sistema de Dos Híbridos
14.
Front Plant Sci ; 11: 584471, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-33154763

RESUMEN

Mitogen-activated protein kinases (MAPK) play pivotal roles in transducing developmental cues and environmental signals into cellular responses through pathways initiated by MAPK kinase kinases (MAP3K). AtYODA is a MAP3K of Arabidopsis thaliana that controls stomatal development and non-canonical immune responses. Arabidopsis plants overexpressing a constitutively active YODA protein (AtCA-YDA) show broad-spectrum disease resistance and constitutive expression of defensive genes. We tested YDA function in crops immunity by heterologously overexpressing AtCA-YDA in Solanum lycopersicum. We found that these tomato AtCA-YDA plants do not show developmental phenotypes and fitness alterations, except a reduction in stomatal index, as reported in Arabidopsis AtCA-YDA plants. Notably, AtCA-YDA tomato plants show enhanced resistance to the bacterial pathogen Pseudomonas syringae pv. tomato DC3000 and constitutive upregulation of defense-associated genes, corroborating the functionality of YDA in tomato immunity. This function was further supported by generating CRISPR/Cas9-edited tomato mutants impaired in the closest orthologs of AtYDA [Solyc08g081210 (SlYDA1) and Solyc03g025360 (SlYDA2)]. Slyda1 and Slyda2 mutants are highly susceptible to P. syringae pv. tomato DC3000 in comparison to wild-type plants but only Slyda2 shows altered stomatal index. These results indicate that tomato orthologs have specialized functions and support that YDA also regulates immune responses in tomato and may be a trait for breeding disease resistance.

15.
Mol Plant Microbe Interact ; 22(8): 953-63, 2009 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-19589071

RESUMEN

Some receptor-like kinases (RLK) control plant development while others regulate immunity. The Arabidopsis ERECTA (ER) RLK regulates both biological processes. To discover specific components of ER-mediated immunity, a genetic screen was conducted to identify suppressors of erecta (ser) susceptibility to Plectosphaerella cucumerina fungus. The ser1 and ser2 mutations restored disease resistance to this pathogen to wild-type levels in the er-1 background but failed to suppress er-associated developmental phenotypes. The deposition of callose upon P. cucumerina inoculation, which was impaired in the er-1 plants, was also restored to near wild-type levels in the ser er-1 mutants. Analyses of er cell walls revealed that total neutral sugars were reduced and uronic acids increased relative to those of wild-type walls. Interestingly, in the ser er-1 walls, neutral sugars were elevated and uronic acids were reduced relative to both er-1 and wild-type plants. The cell-wall changes found in er-1 and the ser er-1 mutants are unlikely to contribute to their developmental alterations. However, they may influence disease resistance, as a positive correlation was found between uronic acids content and resistance to P. cucumerina. We propose a specific function for ER in regulating cell wall-mediated disease resistance that is distinct from its role in development.


Asunto(s)
Proteínas de Arabidopsis/fisiología , Arabidopsis/microbiología , Pared Celular/fisiología , Phyllachorales/fisiología , Proteínas Serina-Treonina Quinasas/fisiología , Receptores de Superficie Celular/fisiología , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Pared Celular/metabolismo , Glucanos/metabolismo , Inmunidad Innata , Mutación , Fenotipo , Hojas de la Planta/genética , Hojas de la Planta/metabolismo , Hojas de la Planta/microbiología , Proteínas Serina-Treonina Quinasas/genética , Proteínas Serina-Treonina Quinasas/metabolismo , Receptores de Superficie Celular/genética , Receptores de Superficie Celular/metabolismo , Ácidos Urónicos/metabolismo
16.
Front Plant Sci ; 7: 897, 2016.
Artículo en Inglés | MEDLINE | ID: mdl-27446127

RESUMEN

ERECTA (ER) receptor-like kinase (RLK) regulates Arabidopsis thaliana organ growth, and inflorescence and stomatal development by interacting with the ERECTA-family genes (ERf) paralogs, ER-like 1 (ERL1) and ERL2, and the receptor-like protein (RLP) TOO MANY MOUTHS (TMM). ER also controls immune responses and resistance to pathogens such as the bacterium Pseudomonas syringae pv. tomato DC3000 (Pto) and the necrotrophic fungus Plectosphaerella cucumerina BMM (PcBMM). We found that er null-mutant plants overexpressing an ER dominant-negative version lacking the cytoplasmic kinase domain (ERΔK) showed an enhanced susceptibility to PcBMM, suggesting that ERΔK associates and forms inactive complexes with additional RLKs/RLPs required for PcBMM resistance. Genetic analyses demonstrated that ER acts in a combinatorial specific manner with ERL1, ERL2, and TMM to control PcBMM resistance. Moreover, BAK1 (BRASSINOSTEROID INSENSITIVE 1-associated kinase 1) RLK, which together with ERf/TMM regulates stomatal patterning and resistance to Pto, was also found to have an unequal contribution with ER in regulating immune responses and resistance to PcBMM. Co-immunoprecipitation experiments in Nicotiana benthamiana further demonstrated BAK1-ER protein interaction. The secreted epidermal pattern factor peptides (EPF1 and EPF2), which are perceived by ERf members to specify stomatal patterning, do not seem to regulate ER-mediated immunity to PcBMM, since their inducible overexpression in A. thaliana did not impact on PcBMM resistance. Our results indicate that the multiproteic receptorsome formed by ERf, TMM and BAK1 modulates A. thaliana resistance to PcBMM, and suggest that the cues underlying ERf/TMM/BAK1-mediated immune responses are distinct from those regulating stomatal pattering.

17.
Mol Plant ; 5(1): 98-114, 2012 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-21980142

RESUMEN

The Arabidopsis heterotrimeric G-protein controls defense responses to necrotrophic and vascular fungi. The agb1 mutant impaired in the Gß subunit displays enhanced susceptibility to these pathogens. Gß/AGB1 forms an obligate dimer with either one of the Arabidopsis Gγ subunits (γ1/AGG1 and γ2/AGG2). Accordingly, we now demonstrate that the agg1 agg2 double mutant is as susceptible as agb1 plants to the necrotrophic fungus Plectosphaerella cucumerina. To elucidate the molecular basis of heterotrimeric G-protein-mediated resistance, we performed a comparative transcriptomic analysis of agb1-1 mutant and wild-type plants upon inoculation with P. cucumerina. This analysis, together with metabolomic studies, demonstrated that G-protein-mediated resistance was independent of defensive pathways required for resistance to necrotrophic fungi, such as the salicylic acid, jasmonic acid, ethylene, abscisic acid, and tryptophan-derived metabolites signaling, as these pathways were not impaired in agb1 and agg1 agg2 mutants. Notably, many mis-regulated genes in agb1 plants were related with cell wall functions, which was also the case in agg1 agg2 mutant. Biochemical analyses and Fourier Transform InfraRed (FTIR) spectroscopy of cell walls from G-protein mutants revealed that the xylose content was lower in agb1 and agg1 agg2 mutants than in wild-type plants, and that mutant walls had similar FTIR spectratypes, which differed from that of wild-type plants. The data presented here suggest a canonical functionality of the Gß and Gγ1/γ2 subunits in the control of Arabidopsis immune responses and the regulation of cell wall composition.


Asunto(s)
Proteínas de Arabidopsis/inmunología , Arabidopsis/inmunología , Ascomicetos/fisiología , Pared Celular/inmunología , Subunidades beta de la Proteína de Unión al GTP/inmunología , Subunidades gamma de la Proteína de Unión al GTP/inmunología , Enfermedades de las Plantas/microbiología , Arabidopsis/química , Arabidopsis/genética , Arabidopsis/microbiología , Proteínas de Arabidopsis/genética , Ascomicetos/inmunología , Pared Celular/química , Pared Celular/genética , Pared Celular/microbiología , Dimerización , Resistencia a la Enfermedad , Subunidades beta de la Proteína de Unión al GTP/genética , Subunidades gamma de la Proteína de Unión al GTP/genética , Regulación de la Expresión Génica de las Plantas , Enfermedades de las Plantas/inmunología
18.
Plant Physiol ; 150(2): 670-83, 2009 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-19363092

RESUMEN

Glutathione peroxidases (GPXs; EC 1.11.1.9) are key enzymes of the antioxidant network in plants and animals. In order to investigate the role of antioxidant systems in plant chloroplasts, we generated Arabidopsis (Arabidopsis thaliana) transgenic lines that are depleted specifically in chloroplastic (cp) forms of GPX1 and GPX7. We show that reduced cpGPX expression, either in transgenic lines with lower total cpGPX expression (GPX1 and GPX7) or in a gpx7 insertion mutant, leads to compromised photooxidative stress tolerance but increased basal resistance to virulent bacteria. Depletion of both GPX1 and GPX7 expression also caused alterations in leaf cell and chloroplast morphology. Leaf tissues were characterized by shorter and more rounded palisade cells, irregular spongy mesophyll cells, and larger intercellular air spaces compared with the wild type. Chloroplasts had larger and more abundant starch grains than in wild-type and gpx7 mutant plants. Constitutively reduced cpGPX expression also led to higher foliar ascorbic acid, glutathione, and salicylic acid levels in plants exposed to higher light intensities. Our results suggest partially overlapping functions of GPX1 and GPX7. The data further point to specific changes in the chloroplast ascorbate-glutathione cycle due to reduced cpGPX expression, initiating reactive oxygen species and salicylic acid pathways that affect leaf development, light acclimation, basal defense, and cell death programs. Thus, cpGPXs regulate cellular photooxidative tolerance and immune responses.


Asunto(s)
Arabidopsis/enzimología , Arabidopsis/inmunología , Cloroplastos/enzimología , Glutatión Peroxidasa/metabolismo , Luz , Estrés Oxidativo/efectos de la radiación , Adaptación Fisiológica/efectos de la radiación , Antioxidantes/metabolismo , Arabidopsis/citología , Arabidopsis/genética , Muerte Celular/efectos de la radiación , Cloroplastos/efectos de la radiación , Cloroplastos/ultraestructura , Perfilación de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Genes de Plantas , Glutatión Peroxidasa/genética , Peróxido de Hidrógeno/metabolismo , Mutación/genética , Hojas de la Planta/citología , Hojas de la Planta/enzimología , Hojas de la Planta/efectos de la radiación , Hojas de la Planta/ultraestructura , Plantas Modificadas Genéticamente , Ácido Salicílico/metabolismo , Estrés Fisiológico/efectos de la radiación
19.
Mol Plant ; 1(3): 496-509, 2008 May.
Artículo en Inglés | MEDLINE | ID: mdl-19825556

RESUMEN

In plants, resistance to necrotrophic pathogens depends on the interplay between different hormone systems, such as those regulated by salicylic acid (SA), jasmonic acid (JA), ethylene, and abscisic acid. Repression of auxin signaling by the SA pathway was recently shown to contribute to antibacterial resistance. Here, we demonstrate that Arabidopsis auxin signaling mutants axr1, axr2, and axr6 that have defects in the auxin-stimulated SCF (Skp1-Cullin-F-box) ubiquitination pathway exhibit increased susceptibility to the necrotrophic fungi Plectosphaerella cucumerina and Botrytis cinerea. Also, stabilization of the auxin transcriptional repressor AXR3 that is normally targeted for removal by the SCF-ubiquitin/proteasome machinery occurs upon P. cucumerina infection. Pharmacological inhibition of auxin transport or proteasome function each compromise necrotroph resistance of wild-type plants to a similar extent as in non-treated auxin response mutants. These results suggest that auxin signaling is important for resistance to the necrotrophic fungi P. cucumerina and B. cinerea. SGT1b (one of two Arabidopsis SGT1 genes encoding HSP90/HSC70 co-chaperones) promotes the functions of SCF E3-ubiquitin ligase complexes in auxin and JA responses and resistance conditioned by certain Resistance (R) genes to biotrophic pathogens. We find that sgt1b mutants are as resistant to P. cucumerina as wild-type plants. Conversely, auxin/SCF signaling mutants are uncompromised in RPP4-triggered resistance to the obligate biotrophic oomycete, Hyaloperonospora parasitica. Thus, the predominant action of SGT1b in R gene-conditioned resistance to oomycetes appears to be at a site other than assisting SCF E3-ubiquitin ligases. However, genetic additivity of sgt1b axr1 double mutants in susceptibility to H. parasitica suggests that SCF-mediated ubiquitination contributes to limiting biotrophic pathogen colonization once plant-pathogen compatibility is established.


Asunto(s)
Arabidopsis/microbiología , Botrytis/patogenicidad , Hongos/patogenicidad , Predisposición Genética a la Enfermedad/epidemiología , Inmunidad Innata/fisiología , Ácidos Indolacéticos/farmacología , Enfermedades de las Plantas/microbiología , Reguladores del Crecimiento de las Plantas/farmacología , Arabidopsis/efectos de los fármacos , Arabidopsis/genética , Proteínas de Arabidopsis/fisiología , Botrytis/efectos de los fármacos , Botrytis/genética , Hongos/efectos de los fármacos , Mutación , Transducción de Señal/efectos de los fármacos , Transducción de Señal/fisiología
20.
Plant J ; 44(1): 155-66, 2005 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-16167903

RESUMEN

We report on constitutive subtilisin3 (csb3), an Arabidopsis mutant showing strikingly enhanced resistance to biotrophic pathogens. Epistasis analyses with pad4, sid2, eds5, NahG, npr1, dth9 and cpr1 mutants revealed that the enhanced resistance of csb3 plants requires intact salicylic acid (SA) synthesis and perception. CSB3 encodes a 1-hydroxy-2-methyl-2-butenyl 4-diphosphate synthase, the enzyme controlling the penultimate step of the biosynthesis of isopentenyl diphosphate via the 2-C-methyl-d-erythritol-4-phosphate (MEP) pathway in the chloroplast. CSB3 is expressed constitutively in healthy plants, and shows repression in response to bacterial infection. We also show the pharmacological complementation of the enhanced-resistance phenotype of csb3 plants with fosmidomycin, an inhibitor of the MEP pathway, and propose that CSB3 represents a point of metabolic convergence modulating the magnitude of SA-mediated disease resistance to biotrophic pathogens.


Asunto(s)
Arabidopsis/genética , Arabidopsis/metabolismo , Enzimas/genética , Enzimas/metabolismo , Eritritol/análogos & derivados , Enfermedades de las Plantas/genética , Ácido Salicílico/metabolismo , Fosfatos de Azúcar/metabolismo , Secuencia de Aminoácidos , Eritritol/metabolismo , Regulación de la Expresión Génica de las Plantas , Datos de Secuencia Molecular , Mutación/genética , Fenotipo , Enfermedades de las Plantas/microbiología , Pseudomonas syringae/fisiología , Homología de Secuencia de Aminoácido , Transducción de Señal
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