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1.
Clin Exp Immunol ; 203(3): 375-384, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-33205391

RESUMO

Despite undeniable improvement in the management of rheumatoid arthritis (RA), the discovery of more effective, less toxic and, ideally, less immune suppressive drugs are much needed. In the current study, we set to explore the potential anti-rheumatic activity of the non-toxic, tellurium-based immunomodulator, AS101 in an experimental animal model of RA. The effect of AS101 was assessed on adjuvant-induced arthritis (AIA) rats. Clinical signs of arthritis were assessed. Histopathological examination was used to assess inflammation, synovial changes and tissue lesions. Very late antigen-4 (VLA-4)+ cellular infiltration was detected using immunohistochemical staining. Enzyme-linked immunosorbent assay (ELISA) was used to measure circulating anti-cyclic citrullinated-peptide autoantibody (ACPA) and real-time polymerase chain reaction (PCR) was used to measure the in-vitro effect of AS101 on interleukin (IL)-6 and IL-1ß expression in activated primary human fibroblasts. Prophylactic treatment with intraperitoneal AS101 reduced clinical arthritis scores in AIA rats (P < 0·01). AS101 abrogated the migration of active chronic inflammatory immune cells, particularly VLA-4+ cells, into joint cartilage and synovium, reduced the extent of joint damage and preserved joint architecture. Compared to phosphate-buffered saline (PBS)-treated AIA rats, histopathological inflammatory scores were significantly reduced (P < 0·05). Furthermore, AS101 resulted in a marked reduction of circulating ACPA in comparison to PBS-treated rats (P < 0·05). Importantly, AS101 significantly reduced mRNA levels of proinflammatory mediators such as IL-6 (P < 0·05) and IL-1ß (P < 0·01) in activated primary human fibroblasts. Taken together, we report the first demonstration of the anti-rheumatic/inflammatory activity of AS101 in experimental RA model, thereby supporting an alternative early therapeutic intervention and identifying a promising agent for therapeutic intervention.


Assuntos
Artrite Experimental/imunologia , Artrite Reumatoide/imunologia , Etilenos/imunologia , Telúrio/imunologia , Adjuvantes Imunológicos/farmacologia , Animais , Artrite Experimental/metabolismo , Artrite Experimental/prevenção & controle , Artrite Reumatoide/metabolismo , Artrite Reumatoide/prevenção & controle , Células Cultivadas , Etilenos/farmacologia , Feminino , Fibroblastos/citologia , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Expressão Gênica/efeitos dos fármacos , Humanos , Fatores Imunológicos/imunologia , Fatores Imunológicos/farmacologia , Integrina alfa4beta1/imunologia , Integrina alfa4beta1/metabolismo , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Interleucina-6/genética , Interleucina-6/metabolismo , Ratos Endogâmicos Lew , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Telúrio/farmacologia
2.
PLoS One ; 14(8): e0221358, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31437216

RESUMO

Plant defense responses to biotic stresses are complex biological processes, all governed by sophisticated molecular regulations. Induced systemic resistance (ISR) is one of these defense mechanisms where beneficial bacteria or fungi prime plants to resist pathogens or pest attacks. In ISR, the defense arsenal in plants remains dormant and it is only triggered by an infection, allowing a better allocation of plant resources. Our group recently described that the well-known beneficial bacterium Paraburkholderia phytofirmans PsJN is able to induce Arabidopsis thaliana resistance to Pseudomonas syringae pv. tomato (Pst) DC3000 through ISR, and that ethylene, jasmonate and salicylic acid are involved in this protection. Nevertheless, the molecular networks governing this beneficial interaction remain unknown. To tackle this issue, we analyzed the temporal changes in the transcriptome of PsJN-inoculated plants before and after being infected with Pst DC3000. These data were used to perform a gene network analysis to identify highly connected transcription factors. Before the pathogen challenge, the strain PsJN regulated 405 genes (corresponding to 1.8% of the analyzed genome). PsJN-inoculated plants presented a faster and stronger transcriptional response at 1-hour post infection (hpi) compared with the non-inoculated plants, which presented the highest transcriptional changes at 24 hpi. A principal component analysis showed that PsJN-induced plant responses to the pathogen could be differentiated from those induced by the pathogen itself. Forty-eight transcription factors were regulated by PsJN at 1 hpi, and a system biology analysis revealed a network with four clusters. Within these clusters LHY, WRKY28, MYB31 and RRTF1 are highly connected transcription factors, which could act as hub regulators in this interaction. Concordantly with our previous results, these clusters are related to jasmonate, ethylene, salicylic, acid and ROS pathways. These results indicate that a rapid and specific response of PsJN-inoculated plants to the virulent DC3000 strain could be the pivotal element in the protection mechanism.


Assuntos
Arabidopsis/genética , Burkholderiaceae/fisiologia , Regulação da Expressão Gênica de Plantas/imunologia , Doenças das Plantas/genética , Pseudomonas syringae/patogenicidade , Fatores de Transcrição/genética , Arabidopsis/imunologia , Arabidopsis/microbiologia , Ciclopentanos/imunologia , Ciclopentanos/metabolismo , Resistência à Doença/genética , Etilenos/imunologia , Etilenos/metabolismo , Perfilação da Expressão Gênica , Redes Reguladoras de Genes/imunologia , Oxilipinas/imunologia , Oxilipinas/metabolismo , Doenças das Plantas/imunologia , Doenças das Plantas/microbiologia , Reguladores de Crescimento de Plantas/imunologia , Reguladores de Crescimento de Plantas/metabolismo , Imunidade Vegetal/genética , Análise de Componente Principal , Pseudomonas syringae/crescimento & desenvolvimento , Ácido Salicílico/imunologia , Ácido Salicílico/metabolismo , Fatores de Transcrição/imunologia , Transcriptoma/imunologia
3.
J Agric Food Chem ; 67(24): 6725-6735, 2019 Jun 19.
Artigo em Inglês | MEDLINE | ID: mdl-31117506

RESUMO

Jasmonic acid (JA)- and ethylene-mediated signaling pathways are reported to have synergistic effects on inhibiting gray mold. The present study aimed to explain the role of ethylene perception in methyl jasmonate (MeJA)-mediated immune responses. Results showed that exogenous MeJA enhanced disease resistance, accompanied by the induction of endogenous JA biosynthesis and ethylene production, which led to the activation of the phenolic metabolism pathway. Blocking ethylene perception using 1-methylcyclopropene (1-MCP) either before or after MeJA treatment could differently weaken the disease responses induced by MeJA, including suppressing the induction of ethylene production and JA contents and reducing activities of lipoxygenase and allene oxide synthase compared to MeJA treatment alone. Consequently, MeJA-induced elevations in the total phenolic content and the activities of phenylalanine ammonia-lyase, cinnamate 4-hydroxylase, 4-coumarate:coenzyme A ligase, and peroxidase were impaired by 1-MCP. These results suggested that ethylene perception participated in MeJA-mediated immune responses in tomato fruit.


Assuntos
Acetatos/imunologia , Botrytis/fisiologia , Ciclopentanos/imunologia , Etilenos/imunologia , Oxilipinas/imunologia , Doenças das Plantas/imunologia , Reguladores de Crescimento de Plantas/imunologia , Solanum lycopersicum/imunologia , Resistência à Doença , Frutas/imunologia , Frutas/microbiologia , Regulação da Expressão Gênica de Plantas , Solanum lycopersicum/genética , Solanum lycopersicum/microbiologia , Fenilalanina Amônia-Liase/genética , Fenilalanina Amônia-Liase/imunologia , Doenças das Plantas/microbiologia , Proteínas de Plantas/genética , Proteínas de Plantas/imunologia , Transcinamato 4-Mono-Oxigenase/genética , Transcinamato 4-Mono-Oxigenase/imunologia
4.
PLoS One ; 13(10): e0205705, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30332458

RESUMO

Phytophthora cinnamomi Rands (Pc) is a hemibiotrophic oomycete and the causal agent of Phytophthora root rot (PRR) of the commercially important fruit crop avocado (Persea americana Mill.). Plant defense against pathogens is modulated by phytohormone signaling pathways such as salicylic acid (SA), jasmonic acid (JA), ethylene (ET), auxin and abscisic acid. The role of specific signaling pathways induced and regulated during hemibiotroph-plant interactions has been widely debated. Some studies report SA mediated defense while others hypothesize that JA responses restrict the spread of pathogens. This study aimed to identify the role of SA- and JA- associated genes in the defense strategy of a resistant avocado rootstock, Dusa in response to Pc infection. Transcripts associated with SA-mediated defense pathways and lignin biosynthesis were upregulated at 6 hours post-inoculation (hpi). Results suggest that auxin, reactive oxygen species (ROS) and Ca2+ signaling was also important during this early time point, while JA signaling was absent. Both SA and JA defense responses were shown to play a role during defense at 18 hpi. Induction of genes associated with ROS detoxification and cell wall digestion (ß-1-3-glucanase) was also observed. Most genes induced at 24 hpi were linked to JA responses. Other processes at play in avocado at 24 hpi include cell wall strengthening, the formation of phenolics and induction of arabinogalactan, a gene linked to Pc zoospore immobility. This study represents the first transcriptome wide analysis of a resistant avocado rootstock treated with SA and JA compared to Pc infection. The results provide evidence of a biphasic defense response against the hemibiotroph, which initially involves SA-mediated gene expression followed by the enrichment of JA-mediated defense from 18 to 24 hpi. Genes and molecular pathways linked to Pc resistance are highlighted and may serve as future targets for manipulation in the development of PRR resistant avocado rootstocks.


Assuntos
Regulação da Expressão Gênica de Plantas/imunologia , Interações Hospedeiro-Patógeno/imunologia , Persea/imunologia , Phytophthora/patogenicidade , Doenças das Plantas/imunologia , Ácido Abscísico/imunologia , Ácido Abscísico/metabolismo , Ciclopentanos/imunologia , Ciclopentanos/metabolismo , Etilenos/imunologia , Etilenos/metabolismo , Perfilação da Expressão Gênica , Interações Hospedeiro-Patógeno/genética , Ácidos Indolacéticos/imunologia , Ácidos Indolacéticos/metabolismo , Oxilipinas/imunologia , Oxilipinas/metabolismo , Persea/genética , Persea/microbiologia , Doenças das Plantas/microbiologia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Raízes de Plantas/genética , Raízes de Plantas/imunologia , Raízes de Plantas/microbiologia , Ácido Salicílico/imunologia , Ácido Salicílico/metabolismo , Transdução de Sinais/genética , Transdução de Sinais/imunologia
5.
Curr Issues Mol Biol ; 26: 55-64, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-28879856

RESUMO

Global crop production is highly threatened due to pathogen invasion. The huge quantity of pesticides application, although harmful to the environment and human health, is carried out to prevent the crop losses worldwide, every year. Therefore, understanding the molecular mechanisms of pathogenicity and plant resistance against pathogen is important. The resistance against pathogens is regulated by three important phytohormones viz. salicylic acid (SA), jasmonic acid (JA) and ethylene (ET). Here we review possible role of CRISPR technology to understand the plant pathogenicity by mutating genes responsible for pathogen invasion or up-regulating the phytohormones genes or resistant genes. Thus hormone biosynthesis genes, receptor and feeding genes of pathogens could be important targets for modifications using CRISPR/Cas9 following multiplexing tool box strategy in order to edit multiple genes simultaneously to produce super plants. Here we put forward our idea thatthe genes would be either mutated in case of plant receptor protein targets of pathogens or up-regulation of resistant genes or hormone biosynthesis genes will be better choice for resistance against pathogens.


Assuntos
Proteínas de Bactérias/genética , Sistemas CRISPR-Cas , Produtos Agrícolas/genética , Resistência à Doença/genética , Endonucleases/genética , Edição de Genes/métodos , Genoma de Planta , Animais , Bactérias/genética , Bactérias/metabolismo , Bactérias/patogenicidade , Proteínas de Bactérias/metabolismo , Proteína 9 Associada à CRISPR , Produtos Agrícolas/imunologia , Produtos Agrícolas/microbiologia , Produtos Agrícolas/parasitologia , Ciclopentanos/imunologia , Ciclopentanos/metabolismo , Endonucleases/metabolismo , Etilenos/biossíntese , Etilenos/imunologia , Fungos/genética , Fungos/metabolismo , Fungos/patogenicidade , Mutação , Nematoides/genética , Nematoides/metabolismo , Nematoides/patogenicidade , Oxilipinas/imunologia , Oxilipinas/metabolismo , Doenças das Plantas/genética , Doenças das Plantas/imunologia , Doenças das Plantas/microbiologia , Doenças das Plantas/parasitologia , Reguladores de Crescimento de Plantas/biossíntese , Reguladores de Crescimento de Plantas/genética , Reguladores de Crescimento de Plantas/imunologia , Proteínas de Plantas/genética , Proteínas de Plantas/imunologia , Ácido Salicílico/imunologia , Ácido Salicílico/metabolismo
6.
PLoS Genet ; 13(5): e1006639, 2017 May.
Artigo em Inglês | MEDLINE | ID: mdl-28472137

RESUMO

Plant immunity protects plants from numerous potentially pathogenic microbes. The biological network that controls plant inducible immunity must function effectively even when network components are targeted and disabled by pathogen effectors. Network buffering could confer this resilience by allowing different parts of the network to compensate for loss of one another's functions. Networks rich in buffering rely on interactions within the network, but these mechanisms are difficult to study by simple genetic means. Through a network reconstitution strategy, in which we disassemble and stepwise reassemble the plant immune network that mediates Pattern-Triggered-Immunity, we have resolved systems-level regulatory mechanisms underlying the Arabidopsis transcriptome response to the immune stimulant flagellin-22 (flg22). These mechanisms show widespread evidence of interactions among major sub-networks-we call these sectors-in the flg22-responsive transcriptome. Many of these interactions result in network buffering. Resolved regulatory mechanisms show unexpected patterns for how the jasmonate (JA), ethylene (ET), phytoalexin-deficient 4 (PAD4), and salicylate (SA) signaling sectors control the transcriptional response to flg22. We demonstrate that many of the regulatory mechanisms we resolved are not detectable by the traditional genetic approach of single-gene null-mutant analysis. Similar to potential pathogenic perturbations, null-mutant effects on immune signaling can be buffered by the network.


Assuntos
Proteínas de Arabidopsis/genética , Hidrolases de Éster Carboxílico/genética , Flagelina/genética , Interações Hospedeiro-Patógeno/genética , Imunidade Vegetal/genética , Transcriptoma/genética , Arabidopsis/genética , Arabidopsis/imunologia , Proteínas de Arabidopsis/imunologia , Hidrolases de Éster Carboxílico/imunologia , Ciclopentanos/imunologia , Ciclopentanos/metabolismo , Etilenos/imunologia , Etilenos/metabolismo , Flagelina/imunologia , Regulação da Expressão Gênica de Plantas , Redes Reguladoras de Genes/imunologia , Interações Hospedeiro-Patógeno/imunologia , Oxilipinas/imunologia , Oxilipinas/metabolismo , Doenças das Plantas/genética , Doenças das Plantas/imunologia , Ácido Salicílico/imunologia , Ácido Salicílico/metabolismo , Transdução de Sinais , Transcriptoma/imunologia
7.
Curr Protein Pept Sci ; 18(4): 294-310, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-27455974

RESUMO

Plants exhibit sensitive mechanisms to respond to environmental stresses, presenting some specific and non-specific reactions when attacked by pathogens, including organisms from different classes and complexity, as viroids, viruses, bacteria, fungi and nematodes. A crucial step to define the fate of the plant facing an invading pathogen is the activation of a compatible Resistance (R) gene, the focus of the present review. Different aspects regarding R-genes and their products are discussed, including pathogen recognition mechanisms, signaling and effects on induced and constitutive defense processes, splicing and post transcriptional mechanisms involved. There are still countless challenges to the complete understanding of the mechanisms involving R-genes in plants, in particular those related to the interactions with other genes of the pathogen and of the host itself, their regulation, acting mechanisms at transcriptional and post-transcriptional levels, as well as the influence of other types of stress over their regulation. A magnification of knowledge is expected when considering the novel information from the omics and systems biology.


Assuntos
Proteínas de Arabidopsis/imunologia , Resistência à Doença/genética , Regulação da Expressão Gênica de Plantas/imunologia , Genoma de Planta , Doenças das Plantas/imunologia , Plantas/genética , Proteínas de Arabidopsis/genética , Mapeamento Cromossômico , Etilenos/biossíntese , Etilenos/imunologia , Dosagem de Genes , Interações Hospedeiro-Patógeno/genética , Interações Hospedeiro-Patógeno/imunologia , Doenças das Plantas/genética , Imunidade Vegetal/genética , Proteínas de Plantas/genética , Proteínas de Plantas/imunologia , Plantas/microbiologia , Plantas/parasitologia , Plantas/virologia , Isoformas de Proteínas/genética , Isoformas de Proteínas/imunologia , Espécies Reativas de Oxigênio/imunologia , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais
8.
Microb Biotechnol ; 7(6): 545-55, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-24888884

RESUMO

Fresh fruits and vegetables are increasingly recognized as important reservoirs of human pathogens, and therefore, significant attention has been directed recently to understanding mechanisms of the interactions between plants and enterics, like Salmonella. A screen of tomato cultivars for their susceptibility to Salmonella revealed significant differences in the ability of this human pathogen to multiply within fruits; expression of the Salmonella genes (cysB, agfB, fadH) involved in the interactions with tomatoes depended on the tomato genotype and maturity stage. Proliferation of Salmonella was strongly reduced in the tomato mutants with defects in ethylene synthesis, perception and signal transduction. While mutation in the ripening-related ethylene receptor Nr resulted only in a modest reduction in Salmonella numbers within tomatoes, strong inhibition of the Salmonella proliferation was observed in rin and nor tomato mutants. RIN and NOR are regulators of ethylene synthesis and ripening. A commercial tomato variety heterozygous for rin was less susceptible to Salmonella under the greenhouse conditions but not when tested in the field over three production seasons.


Assuntos
Etilenos/imunologia , Doenças das Plantas/microbiologia , Salmonella/fisiologia , Solanum lycopersicum/microbiologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Solanum lycopersicum/genética , Solanum lycopersicum/imunologia , Doenças das Plantas/genética , Doenças das Plantas/imunologia , Proteínas de Plantas/genética , Proteínas de Plantas/imunologia , Salmonella/genética
9.
Cell Host Microbe ; 15(1): 84-94, 2014 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-24439900

RESUMO

The plant immune signaling network needs to be robust against attack from fast-evolving pathogens and tunable to optimize immune responses. We investigated the basis of robustness and tunability in the signaling network controlling pattern-triggered immunity (PTI) in Arabidopsis. A dynamic network model containing four major signaling sectors, the jasmonate, ethylene, phytoalexin-deficient 4, and salicylate sectors, which together govern up to 80% of the PTI levels, was built using data for dynamic sector activities and PTI levels under exhaustive combinatorial sector perturbations. Our regularized multiple regression model had a high level of predictive power and captured known and unexpected signal flows in the network. The sole inhibitory sector in the model, the ethylene sector, contributed centrally to network robustness via its inhibition of the jasmonate sector. The model's multiple input sites linked specific signal input patterns varying in strength and timing to different network response patterns, indicating a mechanism enabling tunability.


Assuntos
Proteínas de Arabidopsis/imunologia , Arabidopsis/imunologia , Hidrolases de Éster Carboxílico/imunologia , Ciclopentanos/imunologia , Etilenos/imunologia , Regulação da Expressão Gênica de Plantas/imunologia , Oxilipinas/imunologia , Doenças das Plantas/imunologia , Ácido Salicílico/imunologia , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Hidrolases de Éster Carboxílico/genética , Hidrolases de Éster Carboxílico/metabolismo , Quitosana/imunologia , Quitosana/metabolismo , Ciclopentanos/metabolismo , Etilenos/metabolismo , Modelos Biológicos , Oxilipinas/metabolismo , Doenças das Plantas/genética , Imunidade Vegetal , Proteínas Quinases/genética , Proteínas Quinases/imunologia , Pseudomonas syringae/crescimento & desenvolvimento , Análise de Regressão , Ácido Salicílico/metabolismo , Transdução de Sinais
10.
Plant Sci ; 210: 128-40, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23849120

RESUMO

Rising atmospheric CO2 concentrations can affect the induced defense of plants against herbivory by chewing insects, but little is known about whether elevated CO2 can change the inducible defense of plants against herbivory by aphids, which are phloem-sucking rather than tissue-chewing insects. Interactions between the green peach aphid Myzus persicae and four isogenic Arabidopsis thaliana genotypes including wild type and three induced defense pathway deficient mutants were examined under ambient and elevated CO2. Our data showed that elevated CO2 increased the population abundance of peach aphid when reared on wild type and SA-deficient mutant plants. Regardless of aphid infestation, elevated CO2 decreased the jasmonic acid (JA) but increased the salicylic acid (SA) level in wild-type plants. In addition, elevated CO2 increased SA level in SA-deficient mutant while did not change the JA level in JA-deficient mutant. Pathway enrichment analysis based on high-throughput transcriptome sequencing suggested that CO2 level, aphid infestation, and their interactions (respectively) altered plant defense pathways. Furthermore, qPCR results showed that elevated CO2 up-regulated the expression of SA-dependent defense genes but down-regulated the expression of JA/ethylene-dependent defense genes in wild-type plants infested by aphids. The current study indicated that elevated CO2 tended to enhance the ineffective defense-SA signaling pathway and to reduce the effective defense-JA signaling pathway against aphids, which resulted in increased aphid numbers.


Assuntos
Afídeos/fisiologia , Arabidopsis/efeitos dos fármacos , Dióxido de Carbono/farmacologia , Doenças das Plantas/imunologia , Reguladores de Crescimento de Plantas/metabolismo , Imunidade Vegetal/efeitos dos fármacos , Animais , Afídeos/crescimento & desenvolvimento , Arabidopsis/genética , Arabidopsis/imunologia , Arabidopsis/parasitologia , Proteínas de Arabidopsis/genética , Ciclopentanos/imunologia , Ciclopentanos/metabolismo , Regulação para Baixo , Etilenos/imunologia , Etilenos/metabolismo , Feminino , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Interações Hospedeiro-Parasita , Mutação , Oxilipinas/imunologia , Oxilipinas/metabolismo , Doenças das Plantas/parasitologia , Reguladores de Crescimento de Plantas/imunologia , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/genética , Folhas de Planta/imunologia , Folhas de Planta/parasitologia , Ácido Salicílico/imunologia , Ácido Salicílico/metabolismo , Transdução de Sinais , Transcriptoma , Regulação para Cima
11.
Cell Host Microbe ; 13(2): 143-54, 2013 Feb 13.
Artigo em Inglês | MEDLINE | ID: mdl-23414755

RESUMO

XopD, a type III secretion effector from Xanthomonas euvesicatoria (Xcv), the causal agent of bacterial spot of tomato, is required for pathogen growth and delay of host symptom development. XopD carries a C-terminal SUMO protease domain, a host range determining nonspecific DNA-binding domain and two EAR motifs typically found in repressors of stress-induced transcription. The precise target(s) and mechanism(s) of XopD are obscure. We report that XopD directly targets the tomato ethylene responsive transcription factor SlERF4 to suppress ethylene production, which is required for anti-Xcv immunity and symptom development. SlERF4 expression was required for Xcv ΔxopD-induced ethylene production and ethylene-stimulated immunity. XopD colocalized with SlERF4 in subnuclear foci and catalyzed SUMO1 hydrolysis from lysine 53 of SlERF4, causing SlERF4 destabilization. Mutation of lysine 53 prevented SlERF4 sumoylation, decreased SlERF4 levels, and reduced SlERF4 transcription. These data suggest that XopD desumoylates SlERF4 to repress ethylene-induced transcription required for anti-Xcv immunity.


Assuntos
Etilenos/biossíntese , Proteínas de Plantas/metabolismo , Solanum lycopersicum/microbiologia , Fatores de Transcrição/metabolismo , Xanthomonas/patogenicidade , Proteínas de Bactérias/genética , Proteínas de Bactérias/imunologia , Proteínas de Bactérias/metabolismo , Sistemas de Secreção Bacterianos , Suscetibilidade a Doenças/imunologia , Suscetibilidade a Doenças/microbiologia , Etilenos/imunologia , Genes de Plantas , Interações Hospedeiro-Patógeno , Liases/genética , Liases/metabolismo , Solanum lycopersicum/enzimologia , Solanum lycopersicum/genética , Solanum lycopersicum/imunologia , Doenças das Plantas/imunologia , Doenças das Plantas/microbiologia , Folhas de Planta/microbiologia , Proteínas de Plantas/genética , Complexo de Endopeptidases do Proteassoma/metabolismo , Mapeamento de Interação de Proteínas , Estabilidade Proteica , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Sumoilação , Fatores de Transcrição/genética , Transcrição Gênica , Xanthomonas/crescimento & desenvolvimento , Xanthomonas/imunologia
13.
Microb Ecol ; 65(3): 661-70, 2013 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-23242136

RESUMO

Plant growth-promoting bacteria (PGPB) affect plant cellular processes in various ways. The endophytic bacterial strain Enterobacter radicincitans DSM 16656 has been shown to improve plant growth and yield in various agricultural and vegetable crops. Besides its ability to fix atmospheric nitrogen, produce phytohormones, and solubilize phosphate compounds, the strain is highly competitive against native endophytic organisms and colonizes the endorhizosphere in high numbers. Here, we show that E. radicincitans inoculation of the noncrop plant Arabidopsis thaliana promotes plant growth. Furthermore, high performance liquid chromatography (HPLC) analysis revealed that bacterial inoculation slightly decreased amounts of aliphatic glucosinolates in plant leaves in a fast-growing stage but increased these compounds in an older phase where growth is mostly completed. This effect seems to correlate with developmental stage and depends on the nitrogen requirement. Additionally, nitrogen deficiency studies with seedlings grown on medium containing different nitrogen concentrations suggest that plant nitrogen demand can influence the intensity of plant growth enhancement by E. radicincitans. This endophyte seems not to activate stress-inducible mitogen-activated protein kinases (MAPKs). Analyzing transcription of the defense-related genes PR1, PR2, PR5, and PDF1.2 by quantitative real time polymerase chain reaction (qPCR) revealed that E. radicincitans DSM 16656 is able to induce priming via salicylic acid (SA) or jasmonate (JA)/ethylene (ET) signaling pathways to protect plants against potential pathogen attack.


Assuntos
Arabidopsis/crescimento & desenvolvimento , Arabidopsis/microbiologia , Endófitos/fisiologia , Enterobacter/fisiologia , Glucosinolatos/imunologia , Arabidopsis/genética , Arabidopsis/imunologia , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/imunologia , Ciclopentanos/imunologia , Etilenos/imunologia , Regulação da Expressão Gênica de Plantas , Oxilipinas/imunologia , Reguladores de Crescimento de Plantas/imunologia , Ácido Salicílico/imunologia
14.
J Exp Bot ; 64(2): 637-50, 2013 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-23264518

RESUMO

Three phytohormone molecules - ethylene (ET), jasmonic acid (JA) and salicylic acid (SA) - play key roles in mediating disease response to necrotrophic fungal pathogens. This study investigated the roles of the ET, JA, and SA pathways as well as their crosstalk during the interaction between tomato (Solanum lycopersicum) plants and a necrotrophic fungal pathogen Alternaria alternata f. sp. lycopersici (AAL). Both the ET and JASMONIC ACID INSENSITIVE1 (JAI1) receptor-dependent JA signalling pathways are necessary for susceptibility, while SA response promotes resistance to AAL infection. In addition, the role of JA in susceptibility to AAL is partly dependent on ET biosynthesis and perception, while the SA pathway enhances resistance to AAL and antagonizes the ET response. Based on these results, it is proposed that ET, JA, and SA each on their own can influence the susceptibility of tomato to AAL. Furthermore, the functions of JA and SA in susceptibility to the pathogen are correlated with the enhanced or decreased action of ET, respectively. This study has revealed the functional relationship among the three key hormone pathways in tomato defence against AAL.


Assuntos
Alternaria/fisiologia , Ciclopentanos/imunologia , Etilenos/imunologia , Oxilipinas/imunologia , Doenças das Plantas/microbiologia , Reguladores de Crescimento de Plantas/imunologia , Ácido Salicílico/imunologia , Transdução de Sinais , Solanum lycopersicum/imunologia , Solanum lycopersicum/microbiologia , Doenças das Plantas/imunologia
15.
Anal Biochem ; 314(1): 108-15, 2003 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-12633608

RESUMO

NAD-dependent ADP-ribosylation is one of the posttranslational protein modifications. On mammalian cells, glycosylphosphatidylinositol-anchored cell surface ADP-ribosyltransferases (ARTs) ADP-ribosylate other cell surface proteins and thereby affect important cellular functions. Here we describe convenient flow-cytometric and immunoblot assays for monitoring ADP-ribosylation of cell surface proteins on living cells by exploiting the capacity of ARTs to utilize etheno-NAD as substrate. Etheno-ADP-ribosylation of cell surface proteins can be detected by flow cytometry with 1G4, a monoclonal antibody specific for ethenoadenosine. Labeling of cells with 1G4 is dependent on the expression of cell surface ARTs and occurs only after incubation of ART-expressing cells with etheno-NAD and not with etheno-ADP-ribose. Dose-response analyses show efficient 1G4 staining of ART-expressing cells at micromolar etheno-NAD concentrations. Half-maximal staining is obtained with 1-2 micro M etheno-NAD, saturation is reached at 5-20 micro M etheno-NAD. Immunoblot analyses confirm that ART-expressing cells incorporate ethenoadenosine covalently (i.e., SDS resistant) into several cell surface proteins. The flow-cytometric 1G4 staining assay can be used to identify subpopulations of cells expressing cell surface ART activity and to select ART(hi) cell variants. The immunoblot 1G4 staining assay can also be used to identify etheno-ADP-ribosylated target proteins. These new assays hold promise for many interesting applications in biochemistry and cell biology.


Assuntos
ADP Ribose Transferases/metabolismo , Adenosina/análise , Anticorpos Monoclonais/imunologia , Etilenos/análise , Citometria de Fluxo/métodos , Immunoblotting/métodos , ADP Ribose Transferases/genética , Adenosina/imunologia , Animais , Etilenos/imunologia , Humanos , Proteínas de Membrana/análise , Proteínas de Membrana/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Células Tumorais Cultivadas
16.
Pept Res ; 9(5): 248-53, 1996.
Artigo em Inglês | MEDLINE | ID: mdl-9000251

RESUMO

To probe the interactions between major histocompatibility class-II molecules and the amide bonds of the antigenic peptide main chain, we synthesized ethylenic and reduced analogues of HEL(52-61), an immunogenic peptide for murine major histocompatibility class-II IA k restricted T-cell clones. The synthesis of the corresponding ethylenic analogue of HEL(52-61) in position 53-54 was performed by coupling the Fmoc-protected tripeptide Asp-Tyr-psi [E, CH = CH]Gly with HEL(55-61). Biological tests showed that the ethylenic peptide was presented by major histocompatibility class-II IA kappa molecule and recognized by HEL(52-61)-specific T-cell clones. The corresponding reduced peptide of HEL(52-61) at position 53-54 neither stimulated T-cell clones nor competed with the natural peptide. These results show that, while reduced pseudopeptides might not be appropriate, ethylenic pseudopeptides may be used as probes to dissect the role of hydrogen bonding between the peptide main chain and MHC residues and also help in the design of more stable immunogenic peptides.


Assuntos
Etilenos/imunologia , Antígenos de Histocompatibilidade Classe II/imunologia , Muramidase/química , Muramidase/imunologia , Fragmentos de Peptídeos/química , Fragmentos de Peptídeos/imunologia , Linfócitos T/imunologia , Animais , Células Clonais , Ativação Linfocitária/imunologia , Camundongos , Linfócitos T/citologia
17.
Nat Immun ; 12(1): 50-5, 1993.
Artigo em Inglês | MEDLINE | ID: mdl-8094304

RESUMO

Peripheral blood lymphocytes were obtained from eight untreated patients suffering from long-standing plaque-type psoriasis 1 h following UVB treatment and at the end of 15 UVB treatments. Before treatment and following in vivo treatment with 0.01-0.3 J/cm2, the patients' lymphocytes were tested for interleukin-2 (IL-2) production, and helper activity with and without addition of the immunomodulator AS101. AS101 is a synthetic organotellurium compound, ammonium trichloro-(O,O'-dioxyethylene) tellurate. UVB at a flow of 0.01 J/cm2 did not affect any of these functions. However, after 15 UVB treatments, when the dose given had reached 0.3 J/cm2, a complete inhibition of IL-2 production and helper activity was observed. In vitro addition of AS101 at 0.1 micrograms/ml to the in vivo UVB-treated lymphocytes raised IL-2 production to approximately double the initial level, and improved the helper activity of CD4 cells.


Assuntos
Adjuvantes Imunológicos , Etilenos/imunologia , Interleucina-2/biossíntese , Psoríase/imunologia , Psoríase/radioterapia , Linfócitos T Auxiliares-Indutores/imunologia , Terapia Ultravioleta , Idoso , Animais , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/efeitos da radiação , Feminino , Reação Enxerto-Hospedeiro/imunologia , Humanos , Masculino , Pessoa de Meia-Idade , Ratos , Ratos Endogâmicos Lew , Linfócitos T Auxiliares-Indutores/efeitos da radiação , Telúrio
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