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1.
Front Microbiol ; 13: 1083270, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36583055

RESUMO

Phosphorus (as phosphate, Pi) and iron (Fe) are critical nutrients in plants that are often poorly available in the soil and can be microbially affected. This work aimed to evaluate how plant-rhizobacteria interaction changes due to different Pi or Fe nutritional scenarios and to study the underlying molecular mechanisms of the microbial modulation of these nutrients in plants. Thus, three proteobacteria (Paraburkholderia phytofirmans PsJN, Azospirillum brasilense Sp7, and Pseudomonas putida KT2440) were used to inoculate Arabidopsis seeds. Additionally, the seeds were exposed to a nutritional factor with the following levels for each nutrient: sufficient (control) or low concentrations of a highly soluble source or sufficient concentrations of a low solubility source. Then, the effects of the combinatorial factors were assessed in plant growth, nutrition, and genetic regulation. Interestingly, some bacterial effects in plants depended on the nutrient source (e.g., increased aerial zones induced by the strains), and others (e.g., decreased primary roots induced by Sp7 or KT2440) occurred regardless of the nutritional treatment. In the short-term, PsJN had detrimental effects on plant growth in the presence of the low-solubility Fe compound, but this was not observed in later stages of plant development. A thorough regulation of the phosphorus content was detected in plants independent of the nutritional treatment. Nevertheless, inoculation with KT2440 increased P content by 29% Pi-deficiency exposed plants. Conversely, the inoculation tended to decrease the Fe content in plants, suggesting a competition for this nutrient in the rhizosphere. The P-source also affected the effects of the PsJN strain in a double mutant of the phosphate starvation response (PSR). Furthermore, depending on the nutrient source, PsJN and Sp7 strains differentially regulated PSR and IAA- associated genes, indicating a role of these pathways in the observed differential phenotypical responses. In the case of iron, PsJN and SP7 regulated iron uptake-related genes regardless of the iron source, which may explain the lower Fe content in inoculated plants. Overall, the plant responses to these proteobacteria were not only influenced by the nutrient concentrations but also by their availabilities, the elapsed time of the interaction, and the specific identities of the beneficial bacteria. Graphical AbstractThe effects of the different nutritional and inoculation treatments are indicated for plant growth parameters (A), gene regulation (B) and phosphorus and iron content (C). Figures created with BioRender.com with an academic license.

2.
Int J Mol Sci ; 23(7)2022 Apr 02.
Artigo em Inglês | MEDLINE | ID: mdl-35409333

RESUMO

Class III peroxidases constitute a plant-specific multigene family, where 73 genes have been identified in Arabidopsis thaliana. These genes are members of the reactive oxygen species (ROS) regulatory network in the whole plant, but more importantly, at the root level. In response to abiotic stresses such as cold, heat, and salinity, their expression is significantly modified. To learn more about their transcriptional regulation, an integrative phenotypic, genomic, and transcriptomic study was executed on the roots of A. thaliana Pyrenean populations. Initially, the root phenotyping highlighted 3 Pyrenean populations to be tolerant to cold (Eaux), heat (Herr), and salt (Grip) stresses. Then, the RNA-seq analyses on these three populations, in addition to Col-0, displayed variations in CIII Prxs expression under stressful treatments and between different genotypes. Consequently, several CIII Prxs were particularly upregulated in the tolerant populations, suggesting novel and specific roles of these genes in plant tolerance against abiotic stresses.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Regulação da Expressão Gênica de Plantas , Peroxidases/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Estresse Fisiológico/genética
3.
iScience ; 24(1): 101978, 2021 Jan 22.
Artigo em Inglês | MEDLINE | ID: mdl-33490891

RESUMO

Reactive oxygen species (ROS) are naturally produced by several redox reactions during plant regular metabolism such as photosynthesis and respiration. Due to their chemical properties and high reactivity, ROS were initially described as detrimental for cells during oxidative stress. However, they have been further recognized as key players in numerous developmental and physiological processes throughout the plant life cycle. Recent studies report the important role of ROS as growth regulators during plant root developmental processes such as in meristem maintenance, in root elongation, and in lateral root, root hair, endodermis, and vascular tissue differentiation. All involve multifaceted interplays between steady-state levels of ROS with transcriptional regulators, phytohormones, and nutrients. In this review, we attempt to summarize recent findings about how ROS are involved in multiple stages of plant root development during cell proliferation, elongation, and differentiation.

4.
Acta otorrinolaringol. esp ; 71(2): 99-118, mar.-abr. 2020. tab, ilus
Artigo em Espanhol | IBECS | ID: ibc-192447

RESUMO

La parálisis de Bell es la forma más común de paresia o parálisis facial. Sin embargo, no todos los pacientes con parálisis facial tienen una parálisis de Bell. Otras causas frecuentes incluyen las secuelas del tratamiento del neurinoma del VIII par, el cáncer de cabeza y cuello, la iatrogenia, el zóster ótico y los traumatismos. El abordaje de cada una de estas situaciones es totalmente diferente. El objetivo de esta guía es servir de consejo para el tratamiento y el seguimiento de los pacientes con parálisis facial. Nuestra idea es que la guía sea práctica, haciendo hincapié en recomendaciones efectivas y útiles en el manejo diario de los pacientes. Esta guía ha sido promovida por la Sociedad Española de ORL y escrita por médicos con experiencia en la enfermedad del nervio facial, incluyendo al menos un especialista de cada comunidad autónoma. Redactada en un formato de preguntas y respuestas, incluye 56 cuestiones relevantes relacionadas con el nervio facial


Bell's palsy is the most common diagnosis associated with facial nerve weakness or paralysis. However, not all patients with facial paresis/paralysis have Bell's palsy. Other common causes include treatment of vestibular schwannoma, head and neck tumours, iatrogenic injuries, Herpes zoster, or trauma. The approach to each of these conditions varies widely. The purpose of this guideline is to provide clinicians with guidance on the treatment and monitoring of patients with different causes of facial paralysis. We intend to draft a practical guideline, focusing on operationalised recommendations deemed to be useful in the daily management of patients. This guideline was promoted by the Spanish Society of Otolaryngology and developed by a group of physicians with an interest in facial nerve disorders, including at least one physician from each Autonomous Community. In a question and answer format, it includes 56 relevant topics related to the facial nerve


Assuntos
Humanos , Paralisia Facial/diagnóstico , Paralisia Facial/terapia , Sociedades Médicas/normas , Otolaringologia/métodos , Paralisia de Bell/etiologia , Espanha , Nervo Facial/fisiopatologia , Paralisia Facial/etiologia
5.
Mol Plant ; 13(5): 698-716, 2020 05 04.
Artigo em Inglês | MEDLINE | ID: mdl-31904511

RESUMO

The molecular links between extracellular signals and the regulation of localized protein synthesis in plant cells are poorly understood. Here, we show that in Arabidopsis thaliana, the extracellular peptide RALF1 and its receptor, the FERONIA receptor kinase, promote root hair (RH) tip growth by modulating protein synthesis. We found that RALF1 promotes FERONIA-mediated phosphorylation of eIF4E1, a eukaryotic translation initiation factor that plays a crucial role in the control of mRNA translation rate. Phosphorylated eIF4E1 increases mRNA affinity and modulates mRNA translation and, thus, protein synthesis. The mRNAs targeted by the RALF1-FERONIA-eIF4E1 module include ROP2 and RSL4, which are important regulators of RH cell polarity and growth. RALF1 and FERONIA are expressed in a polar manner in RHs, which facilitate eIF4E1 polar localization and thus may control local ROP2 translation. Moreover, we demonstrated that high-level accumulation of RSL4 exerts negative-feedback regulation of RALF1 expression by directly binding the RALF1 gene promoter, determining the final RH size. Our study reveals that the link between RALF1-FERONIA signaling and protein synthesis constitutes a novel component regulating cell expansion in these polar growing cells.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/metabolismo , Fator de Iniciação 4E em Eucariotos/metabolismo , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/metabolismo , Biossíntese de Proteínas , Arabidopsis/genética , Tamanho Celular , Retroalimentação Fisiológica , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Tamanho do Órgão , Hormônios Peptídicos/metabolismo , Fosforilação , Fosfotransferases/metabolismo , Regiões Promotoras Genéticas/genética , Ligação Proteica , RNA Mensageiro/genética , RNA Mensageiro/metabolismo
6.
Artigo em Inglês, Espanhol | MEDLINE | ID: mdl-31097197

RESUMO

Bell's palsy is the most common diagnosis associated with facial nerve weakness or paralysis. However, not all patients with facial paresis/paralysis have Bell's palsy. Other common causes include treatment of vestibular schwannoma, head and neck tumours, iatrogenic injuries, Herpes zoster, or trauma. The approach to each of these conditions varies widely. The purpose of this guideline is to provide clinicians with guidance on the treatment and monitoring of patients with different causes of facial paralysis. We intend to draft a practical guideline, focusing on operationalised recommendations deemed to be useful in the daily management of patients. This guideline was promoted by the Spanish Society of Otolaryngology and developed by a group of physicians with an interest in facial nerve disorders, including at least one physician from each Autonomous Community. In a question and answer format, it includes 56 relevant topics related to the facial nerve.


Assuntos
Paralisia de Bell/terapia , Paralisia Facial/terapia , Otolaringologia , Sociedades Médicas , Fatores Etários , Paralisia de Bell/diagnóstico , Diagnóstico Diferencial , Diagnóstico por Imagem , Emergências , Paralisia Facial/diagnóstico , Paralisia Facial/etiologia , Feminino , Humanos , Oftalmologia , Gravidez , Complicações na Gravidez/tratamento farmacológico , Encaminhamento e Consulta , Espanha
7.
Plant Cell Environ ; 41(3): 661-674, 2018 03.
Artigo em Inglês | MEDLINE | ID: mdl-29314044

RESUMO

Abortion of fertilized ovaries at the tip of the ear can generate significant yield losses in maize crops. To investigate the mechanisms involved in this process, 2 maize hybrids were grown in field crops at 2 sowing densities and under 3 irrigation regimes (well-watered control, drought before pollination, and drought during pollination), in all possible combinations. Samples of ear tips were taken 2-6 days after synchronous hand pollination and used for the analysis of gene expression and sugars. Glucose and fructose levels increased in kernels with high abortion risk. Several FASCICLIN-LIKE ARABINOGALACTAN PROTEIN (FLA) genes showed negative correlation with abortion. The expression of ZmFLA7 responded to drought only at the tip of the ear. The abundance of arabinogalactan protein (AGP) glycan epitopes decreased with drought and pharmacological treatments that reduce AGP activity enhanced the abortion of fertilized ovaries. Drought also reduced the expression of AthFLA9 in the siliques of Arabidopsis thaliana. Gain- and loss-of-function mutants of Arabidopsis showed a negative correlation between AthFLA9 and seed abortion. On the basis of gene expression patterns, pharmacological, and genetic evidence, we propose that stress-induced reductions in the expression of selected FLA genes enhance abortion of fertilized ovaries in maize and Arabidopsis.


Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Regulação da Expressão Gênica de Plantas , Mucoproteínas/genética , Proteínas de Plantas/genética , Sementes/fisiologia , Zea mays/genética , Arabidopsis/fisiologia , Proteínas de Arabidopsis/metabolismo , Quimera , Secas , Glucosídeos/farmacologia , Mucoproteínas/metabolismo , Óvulo Vegetal/genética , Óvulo Vegetal/fisiologia , Floroglucinol/análogos & derivados , Floroglucinol/farmacologia , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas , Polinização , Sementes/genética , Zea mays/efeitos dos fármacos , Zea mays/fisiologia
8.
Plant J ; 91(4): 613-630, 2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-28482115

RESUMO

Fasciclin-like arabinogalactan proteins (FLAs) are involved in numerous important functions in plants but the relevance of their complex structure to physiological function and cellular fate is unresolved. Using a fully functional fluorescent version of Arabidopsis thaliana FLA4 we show that this protein is localized at the plasma membrane as well as in endosomes and soluble in the apoplast. FLA4 is likely to be GPI-anchored, is highly N-glycosylated and carries two O-glycan epitopes previously associated with arabinogalactan proteins. The activity of FLA4 was resistant against deletion of the amino-proximal fasciclin 1 domain and was unaffected by removal of the GPI-modification signal, a highly conserved N-glycan or the deletion of predicted O-glycosylation sites. Nonetheless these structural changes dramatically decreased endoplasmic reticulum (ER)-exit and plasma membrane localization of FLA4, with N-glycosylation acting at the level of ER-exit and O-glycosylation influencing post-secretory fate. We show that FLA4 acts predominantly by molecular interactions involving its carboxy-proximal fasciclin 1 domain and that its amino-proximal fasciclin 1 domain is required for stabilization of plasma membrane localization. FLA4 functions as a soluble glycoprotein via its carboxy-proximal Fas1 domain and its normal cellular trafficking depends on N- and O-glycosylation.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Moléculas de Adesão Celular/metabolismo , Arabidopsis/citologia , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Moléculas de Adesão Celular/genética , Retículo Endoplasmático/metabolismo , Glicoproteínas/genética , Glicoproteínas/metabolismo , Glicosilação , Proteínas Luminescentes , Mucoproteínas/genética , Mucoproteínas/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Raízes de Plantas/citologia , Raízes de Plantas/genética , Raízes de Plantas/metabolismo , Polissacarídeos/metabolismo , Domínios Proteicos , Transporte Proteico , Proteínas Recombinantes de Fusão
9.
Proc Natl Acad Sci U S A ; 114(20): 5289-5294, 2017 05 16.
Artigo em Inglês | MEDLINE | ID: mdl-28461488

RESUMO

Root hair polar growth is endogenously controlled by auxin and sustained by oscillating levels of reactive oxygen species (ROS). These cells extend several hundred-fold their original size toward signals important for plant survival. Although their final cell size is of fundamental importance, the molecular mechanisms that control it remain largely unknown. Here we show that ROS production is controlled by the transcription factor RSL4, which in turn is transcriptionally regulated by auxin through several auxin response factors (ARFs). In this manner, auxin controls ROS-mediated polar growth by activating RSL4, which then up-regulates the expression of genes encoding NADPH oxidases (also known as RESPIRATORY BURST OXIDASE HOMOLOG proteins) and class III peroxidases, which catalyze ROS production. Chemical or genetic interference with ROS balance or peroxidase activity affects root hair final cell size. Overall, our findings establish a molecular link between auxin and ROS-mediated polar root hair growth.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Ácidos Indolacéticos/metabolismo , Proteínas de Arabidopsis/genética , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Regulação da Expressão Gênica de Plantas , NADPH Oxidases/metabolismo , Peroxidases/metabolismo , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Fatores de Transcrição/metabolismo
10.
Plant Physiol ; 160(4): 2109-24, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23037505

RESUMO

Plant resistance to necrotrophic fungi is regulated by a complex set of signaling pathways that includes those mediated by the hormones salicylic acid (SA), ethylene (ET), jasmonic acid (JA), and abscisic acid (ABA). The role of ABA in plant resistance remains controversial, as positive and negative regulatory functions have been described depending on the plant-pathogen interaction analyzed. Here, we show that ABA signaling negatively regulates Arabidopsis (Arabidopsis thaliana) resistance to the necrotrophic fungus Plectosphaerella cucumerina. Arabidopsis plants impaired in ABA biosynthesis, such as the aba1-6 mutant, or in ABA signaling, like the quadruple pyr/pyl mutant (pyr1pyl1pyl2pyl4), were more resistant to P. cucumerina than wild-type plants. In contrast, the hab1-1abi1-2abi2-2 mutant impaired in three phosphatases that negatively regulate ABA signaling displayed an enhanced susceptibility phenotype to this fungus. Comparative transcriptomic analyses of aba1-6 and wild-type plants revealed that the ABA pathway negatively regulates defense genes, many of which are controlled by the SA, JA, or ET pathway. In line with these data, we found that aba1-6 resistance to P. cucumerina was partially compromised when the SA, JA, or ET pathway was disrupted in this mutant. Additionally, in the aba1-6 plants, some genes encoding cell wall-related proteins were misregulated. Fourier transform infrared spectroscopy and biochemical analyses of cell walls from aba1-6 and wild-type plants revealed significant differences in their Fourier transform infrared spectratypes and uronic acid and cellulose contents. All these data suggest that ABA signaling has a complex function in Arabidopsis basal resistance, negatively regulating SA/JA/ET-mediated resistance to necrotrophic fungi.


Assuntos
Ácido Abscísico/metabolismo , Arabidopsis/imunologia , Arabidopsis/microbiologia , Ascomicetos/fisiologia , Resistência à Doença/imunologia , Doenças das Plantas/microbiologia , Transdução de Sinais , Arabidopsis/efeitos dos fármacos , Arabidopsis/genética , Ascomicetos/efeitos dos fármacos , Parede Celular/efeitos dos fármacos , Parede Celular/metabolismo , Análise por Conglomerados , Ciclopentanos/metabolismo , Resistência à Doença/efeitos dos fármacos , Resistência à Doença/genética , Etilenos/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Genes de Plantas/genética , Modelos Biológicos , Mutação/genética , Oxilipinas/metabolismo , Doenças das Plantas/genética , Doenças das Plantas/imunologia , Reguladores de Crescimento de Plantas/farmacologia , Ácido Salicílico/metabolismo , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética , Espectroscopia de Infravermelho com Transformada de Fourier , Estresse Fisiológico/efeitos dos fármacos , Estresse Fisiológico/genética
11.
Proc Natl Acad Sci U S A ; 109(11): 4098-103, 2012 Mar 13.
Artigo em Inglês | MEDLINE | ID: mdl-22375033

RESUMO

The mechanisms underlying the biosynthesis of cellulose in plants are complex and still poorly understood. A central question concerns the mechanism of microfibril structure and how this is linked to the catalytic polymerization action of cellulose synthase (CESA). Furthermore, it remains unclear whether modification of cellulose microfibril structure can be achieved genetically, which could be transformative in a bio-based economy. To explore these processes in planta, we developed a chemical genetic toolbox of pharmacological inhibitors and corresponding resistance-conferring point mutations in the C-terminal transmembrane domain region of CESA1(A903V) and CESA3(T942I) in Arabidopsis thaliana. Using (13)C solid-state nuclear magnetic resonance spectroscopy and X-ray diffraction, we show that the cellulose microfibrils displayed reduced width and an additional cellulose C4 peak indicative of a degree of crystallinity that is intermediate between the surface and interior glucans of wild type, suggesting a difference in glucan chain association during microfibril formation. Consistent with measurements of lower microfibril crystallinity, cellulose extracts from mutated CESA1(A903V) and CESA3(T942I) displayed greater saccharification efficiency than wild type. Using live-cell imaging to track fluorescently labeled CESA, we found that these mutants show increased CESA velocities in the plasma membrane, an indication of increased polymerization rate. Collectively, these data suggest that CESA1(A903V) and CESA3(T942I) have modified microfibril structure in terms of crystallinity and suggest that in plants, as in bacteria, crystallization biophysically limits polymerization.


Assuntos
Proteínas de Arabidopsis/química , Proteínas de Arabidopsis/genética , Celulose/química , Glucosiltransferases/química , Glucosiltransferases/genética , Microfibrilas/química , Mutação/genética , Alelos , Sequência de Aminoácidos , Substituição de Aminoácidos/genética , Arabidopsis/efeitos dos fármacos , Arabidopsis/enzimologia , Proteínas de Arabidopsis/metabolismo , Membrana Celular/efeitos dos fármacos , Membrana Celular/enzimologia , Celulose/biossíntese , Cristalização , Resistência a Medicamentos/efeitos dos fármacos , Genes Dominantes/genética , Glucosiltransferases/metabolismo , Espectroscopia de Ressonância Magnética , Microfibrilas/efeitos dos fármacos , Dados de Sequência Molecular , Proteínas Mutantes/química , Proteínas Mutantes/metabolismo , Transporte Proteico/efeitos dos fármacos , Quinolinas/química , Quinolinas/farmacologia , Relação Estrutura-Atividade
12.
Carbohydr Polym ; 87(1): 916-919, 2012 Jan 04.
Artigo em Inglês | MEDLINE | ID: mdl-34663054

RESUMO

ß-(1→4)-d-Mannans constitute the major component of the cell wall of seaweeds of the genus Codium and replace cellulose as the major fibrillar component. They were found as major constituents of the hot water extracts of green seaweed Codium vermilara. By anion exchange chromatography of the first hot water extract, a pure sulfated mannan with a molar ratio carbohydrates:sulfate of 2.7:1 was isolated. The sulfate groups are linked to C-2 of 23% of the mannose units, while most of these units are not substituted. This degree of sulfation would explain the higher solubility of the polymer, compared to that of the non-sulfated fibrillar mannan. Taking into account that the fibrillar polysaccharides form two external layers in the cell wall, while the sulfated polymers are forming an amorphous central layer, it is postulated that these sulfated mannans could act as an interphase region between the neutral and acidic layers.

13.
Mol Plant ; 5(1): 98-114, 2012 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-21980142

RESUMO

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.


Assuntos
Proteínas de Arabidopsis/imunologia , Arabidopsis/imunologia , Ascomicetos/fisiologia , Parede Celular/imunologia , Subunidades beta da Proteína de Ligação ao GTP/imunologia , Subunidades gama da Proteína de Ligação ao GTP/imunologia , Doenças das Plantas/microbiologia , Arabidopsis/química , Arabidopsis/genética , Arabidopsis/microbiologia , Proteínas de Arabidopsis/genética , Ascomicetos/imunologia , Parede Celular/química , Parede Celular/genética , Parede Celular/microbiologia , Dimerização , Resistência à Doença , Subunidades beta da Proteína de Ligação ao GTP/genética , Subunidades gama da Proteína de Ligação ao GTP/genética , Regulação da Expressão Gênica de Plantas , Doenças das Plantas/imunologia
14.
Plant Physiol ; 158(1): 252-63, 2012 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-22080603

RESUMO

Snakin-1 (SN1) is an antimicrobial cysteine-rich peptide isolated from potato (Solanum tuberosum) that was classified as a member of the Snakin/Gibberellic Acid Stimulated in Arabidopsis protein family. In this work, a transgenic approach was used to study the role of SN1 in planta. Even when overexpressing SN1, potato lines did not show remarkable morphological differences from the wild type; SN1 silencing resulted in reduced height, which was accompanied by an overall reduction in leaf size and severe alterations of leaf shape. Analysis of the adaxial epidermis of mature leaves revealed that silenced lines had 70% to 90% increases in mean cell size with respect to wild-type leaves. Consequently, the number of epidermal cells was significantly reduced in these lines. Confocal microscopy analysis after agroinfiltration of Nicotiana benthamiana leaves showed that SN1-green fluorescent protein fusion protein was localized in plasma membrane, and bimolecular fluorescence complementation assays revealed that SN1 self-interacted in vivo. We further focused our study on leaf metabolism by applying a combination of gas chromatography coupled to mass spectrometry, Fourier transform infrared spectroscopy, and spectrophotometric techniques. These targeted analyses allowed a detailed examination of the changes occurring in 46 intermediate compounds from primary metabolic pathways and in seven cell wall constituents. We demonstrated that SN1 silencing affects cell division, leaf primary metabolism, and cell wall composition in potato plants, suggesting that SN1 has additional roles in growth and development beyond its previously assigned role in plant defense.


Assuntos
Parede Celular/química , Proteínas de Plantas/genética , Solanum tuberosum/genética , Solanum tuberosum/metabolismo , Divisão Celular , Membrana Celular/metabolismo , Parede Celular/genética , Parede Celular/metabolismo , Cromatografia Gasosa-Espectrometria de Massas , Regulação da Expressão Gênica de Plantas , Inativação Gênica , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Dados de Sequência Molecular , Epiderme Vegetal/citologia , Epiderme Vegetal/genética , Folhas de Planta/genética , Folhas de Planta/fisiologia , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas , Solanaceae/genética , Solanum tuberosum/citologia , Espectroscopia de Infravermelho com Transformada de Fourier
15.
J Phycol ; 48(2): 326-35, 2012 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-27009722

RESUMO

Bryopsis sp. from a restricted area of the rocky shore of Mar del Plata (Argentina) on the Atlantic coast was identified as Bryopsis plumosa (Hudson) C. Agardh (Bryopsidales, Chlorophyta) based on morphological characters and rbcL and tufA DNA barcodes. To analyze the cell wall polysaccharides of this seaweed, the major room temperature (B1) and 90°C (X1) water extracts were studied. By linkage analysis and NMR spectroscopy, the structure of a sulfated galactan was determined, and putative sulfated rhamnan structures and furanosidic nonsulfated arabinan structures were also found. By anion exchange chromatography of X1, a fraction (F4), comprising a sulfated galactan as major structure was isolated. Structural analysis showed a linear backbone constituted of 3-linked ß-d-galactose units, partially sulfated on C-6 and partially substituted with pyruvic acid forming an acetal linked to O-4 and O-6. This galactan has common structural features with those of green seaweeds of the genus Codium (Bryopsidales, Chlorophyta), but some important differences were also found. This is the first report about the structure of the water-soluble polysaccharides biosynthesized by seaweeds of the genus Bryopsis. These sulfated galactans and rhamnans were in situ localized mostly in two layers, one close to the plasma membrane and the other close to the apoplast, leaving a middle amorphous, unstained cell wall zone. In addition, fibrillar polysaccharides, comprising (1→3)-ß-d-xylans and cellulose, were obtained by treatment of the residue from the water extractions with an LiCl/DMSO solution at high temperature. These polymers were also localized in a bilayer arrangement.

16.
J Phycol ; 47(4): 802-10, 2011 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-27020016

RESUMO

Cell wall chemistry in the coencocytic green seaweed Codium vermilara (Olivi) Delle Chiaje (Bryopsidales, Chlorophyta) is well understood. These cell walls are composed of major amounts of neutral ß-(1→4)-D-mannans (Mn), sulfated polysaccharides (SPs), which include pyranosic arabinan sulfates (ArpS), pyruvylated galactan sulfates (pGaS), and mannan sulfates (MnS); also minor amounts of O-glycoproteins are present. In this study, cell wall samples of C. vermilara were investigated with regard to their monosaccharide composition and infrared spectra (using Fourier transform infrared spectroscopy coupled to principal component [FTIR-PC] analysis). Samples from three different populations of C. vermilara from the Argentine coast showed: (i) an important variation in the relative arabinan content, which increases from north to south, and (ii) a measurable degree of cell wall variability in the sulfate distribution between the different sulfated polysaccharides, independent of the amount of each polysaccharide present and of total sulfate content. When cell wall composition was analyzed over three consecutive years in a single geographic location, the quantity of Mn and overall sulfate content on SPs remained constant, whereas the pGaS:ArpS molar ratio changed over the time. Besides, similar cell wall composition was found between actively growing and resting zones of the thallus, suggesting that cell wall composition is independent of growth stage and development. Overall, these results suggest that C. vermilara has developed a mechanism to adjust the total level of cell wall sulfation by modulating the ArpS:pGaS:MnS molar ratio and also by adjusting the sulfation level in each type of polymer, whereas nonsulfated Mn, as the main structural polysaccharide, did not change over the time or growing stage.

17.
Mol Plant Microbe Interact ; 22(8): 953-63, 2009 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-19589071

RESUMO

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.


Assuntos
Proteínas de Arabidopsis/fisiologia , Arabidopsis/microbiologia , Parede Celular/fisiologia , Phyllachorales/fisiologia , Proteínas Serina-Treonina Quinases/fisiologia , Receptores de Superfície Celular/fisiologia , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Parede Celular/metabolismo , Glucanos/metabolismo , Imunidade Inata , Mutação , Fenótipo , Folhas de Planta/genética , Folhas de Planta/metabolismo , Folhas de Planta/microbiologia , Proteínas Serina-Treonina Quinases/genética , Proteínas Serina-Treonina Quinases/metabolismo , Receptores de Superfície Celular/genética , Receptores de Superfície Celular/metabolismo , Ácidos Urônicos/metabolismo
18.
Glycobiology ; 19(3): 212-28, 2009 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-18832454

RESUMO

A comprehensive analysis of the carbohydrate-containing macromolecules from the coencocytic green seaweed Codium fragile and their arrangement in the cell wall was carried out. Cell walls in this seaweed are highly complex structures composed of 31% (w/w) of linear (1-->4)-beta-D-mannans, 9% (w/w) of pyruvylated arabinogalactan sulfates (pAGS), and low amounts of hydroxyproline rich-glycoprotein epitopes (HRGP). In situ chemical imaging by synchrotron radiation Fourier transform infrared (SR-FTIR) microspectroscopy and by immunolabeling using antibodies against specific cell wall carbohydrate epitopes revealed that beta-d-mannans and pAGS are placed in the middle part of the cell wall, whereas HRGP epitopes (arabinogalactan proteins (AGPs) and extensins) are located on the wall boundaries, especially in the utricle apical zone. pAGS are sulfated at C-2 and/or C-4 of the 3-linked beta-L-arabinopyranose units and at C-4 and/or C-6 of the 3-linked beta-D-galactopyranose residues. In addition, high levels of ketals of pyruvic acid were found mainly at 3,4- of some terminal beta-D-Galp units forming a five-membered ring. Ramification was found at some C-6 of the 3-linked beta-D-Galp units. In agreement with the immunolabeled AGP epitopes, a nonsulfated branched furanosidic arabinan with 5-linked alpha-L-Araf, 3,5-linked alpha-L-Araf, and terminal alpha-L-Araf units and a nonsulfated galactan structure composed of 3-(3,6)-linked beta-D-Galp residues, both typical of type-II AG glycans were found, suggesting that AGP structures are present at low levels in the cell walls of this seaweed. Based on this study, it is starting to emerge that Codium has developed unique cell wall architecture, when compared, not only with that of vascular plants, but also with other related green seaweeds and algae.


Assuntos
Parede Celular/química , Epitopos/análise , Galactanos/análise , Mananas/análise , Alga Marinha/química , Sulfatos/análise , Epitopos/química , Galactanos/química , Galactanos/isolamento & purificação , Hidroxiprolina/análise , Hidroxiprolina/química , Mananas/química , Ressonância Magnética Nuclear Biomolecular , Espectroscopia de Infravermelho com Transformada de Fourier , Sulfatos/química , Síncrotrons
19.
Int J Biol Macromol ; 41(5): 641-9, 2007 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-17920674

RESUMO

Codium fragile and Codium vermilara biosynthesize water-soluble sulfated arabinans and galactans (and/or sulfated arabinogalactans), alpha(1-->4)-D-glucans and beta(1-->4)-D-mannans. The former polysaccharides are composed by 3-linked beta-D-galactopyranose and beta-L-arabinopyranose residues, they are highly sulfated and substituted with pyruvic acid ketals. For both seaweeds, they have the same main structural units, but in different percentages. All the room-temperature water extracts from both seaweeds showed a dual haemostatic effect: they prevented coagulation, but they induced platelet aggregation. Anticoagulant activity and platelet aggregation were higher in the samples with polysaccharides richer in sulfate, mainly in those from C. vermilara, which have a higher degree of sulfation and arabinose content.


Assuntos
Anticoagulantes/farmacologia , Hemostasia/efeitos dos fármacos , Agregação Plaquetária/efeitos dos fármacos , Polissacarídeos/isolamento & purificação , Polissacarídeos/farmacologia , Alga Marinha/química , Arabinose , Galactose , Humanos , Espectroscopia de Ressonância Magnética , Pronase
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