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1.
mBio ; 11(1)2020 02 18.
Artigo em Inglês | MEDLINE | ID: mdl-32071264

RESUMO

The presence of CpG and UpA dinucleotides is restricted in the genomes of animal RNA viruses to avoid specific host defenses. We wondered whether a similar phenomenon exists in nonanimal RNA viruses. Here, we show that these two dinucleotides, especially UpA, are underrepresented in the family Potyviridae, the most important group of plant RNA viruses. Using plum pox virus (PPV; Potyviridae family) as a model, we show that an increase in UpA frequency strongly diminishes virus accumulation. Remarkably, unlike previous observations in animal viruses, PPV variants harboring CpG-rich fragments display just faint (or no) attenuation. The anticorrelation between UpA frequency and viral fitness additionally demonstrates the relevance of this particular dinucleotide: UpA-high mutants are attenuated in a dose-dependent manner, whereas a UpA-low variant displays better fitness than its parental control. Using high-throughput sequencing, we also show that UpA-rich PPV variants are genetically stable, without apparent changes in sequence that revert and/or compensate for the dinucleotide modification despite its attenuation. In addition, we also demonstrate here that the PPV restriction of UpA-rich variants works independently of the classical RNA silencing pathway. Finally, we show that the anticorrelation between UpA frequency and RNA accumulation applies to mRNA-like fragments produced by the host RNA polymerase II. Together, our results inform us about a dinucleotide-based system in plant cells that controls diverse RNAs, including RNA viruses.IMPORTANCE Dinucleotides (combinations of two consecutive nucleotides) are not randomly present in RNA viruses; in fact, the presence of CpG and UpA is significantly repressed in their genomes. Although the meaning of this phenomenon remains obscure, recent studies with animal-infecting viruses have revealed that their low CpG/UpA frequency prevents virus restriction via a host antiviral system that recognizes, and promotes the degradation of, CpG/UpA-rich RNAs. Whether similar systems act in organisms from other life kingdoms has been unknown. To fill this gap in our knowledge, we built several synthetic variants of a plant RNA virus with deoptimized dinucleotide frequencies and analyzed their viral fitness and genome adaptation. In brief, our results inform us for the first time about an effective dinucleotide-based system that acts in plants against viruses. Remarkably, this viral restriction in plants is reminiscent of, but not identical to, the equivalent antiviral response in animals.


Assuntos
Genoma Viral , Doenças das Plantas/virologia , Vírus de Plantas/genética , RNA Viral/genética , Viroses/virologia , Antivirais/farmacologia , Ilhas de CpG/genética , Vírus de DNA/genética , Fosfatos de Dinucleosídeos/genética , Instabilidade Genômica , Doenças das Plantas/imunologia , Vírus de Plantas/efeitos dos fármacos , Vírus Eruptivo da Ameixa , Potyviridae/genética , Potyvirus/genética , Interferência de RNA , RNA Polimerase II , RNA Mensageiro/metabolismo , RNA Viral/imunologia , Viroses/imunologia
2.
J Agric Food Chem ; 68(9): 2631-2638, 2020 Mar 04.
Artigo em Inglês | MEDLINE | ID: mdl-32023057

RESUMO

Previously, we reported for the first time that harmala alkaloids harmine and tetrahydroharmine exhibit activity against plant viruses, and we developed an analogue, designated NK0209, that efficiently prevents and controls plant virus diseases. Here, to investigate the influence of the spatial configuration of NK0209 on its antiviral activities, we synthesized its four optical isomers, determined their configurations, and evaluated their activities against tobacco mosaic virus. All four isomers were significantly more active than ningnanmycin, which is one of the most successful commercial antiviral agents, with in vivo inactivation, cure, and protection rates of 57.3 ± 1.9, 54.2 ± 3.3, and 55.0 ± 4.1% at 500 µg/mL. Furthermore, analysis of structure-activity relationships demonstrated for the first time that the spatial conformation of NK0209 is an important determinant of its antiviral activity, and our results provide information about the possible optimum configuration for interaction of this molecule with its target protein.


Assuntos
Antivirais/química , Antivirais/farmacologia , Harmina/química , Harmina/farmacologia , Vírus de Plantas/efeitos dos fármacos , Vírus do Mosaico do Tabaco/efeitos dos fármacos , Antivirais/síntese química , Desenho de Fármacos , Harmina/síntese química , Isomerismo , Conformação Molecular , Doenças das Plantas/virologia , Vírus de Plantas/crescimento & desenvolvimento , Relação Estrutura-Atividade , Vírus do Mosaico do Tabaco/crescimento & desenvolvimento
3.
J Agric Food Chem ; 67(48): 13344-13352, 2019 Dec 04.
Artigo em Inglês | MEDLINE | ID: mdl-31721573

RESUMO

A series of novel anthranilic diamide derivatives (5a-5ab) containing moieties of trifluoromethylpyridine and hydrazone was designed and synthesized. The synthesized compounds were evaluated in vivo for their activities against tobacco mosaic virus (TMV) and cucumber mosaic virus (CMV). Most of the synthesized compounds displayed good to excellent antiviral activities. The compounds 5i, 5k, 5s, 5w, 5x, and 5z had the curative activity over 65% against TMV at the concentration of 500 µg/mL, which were significantly higher than those of ningnanmycin (55.0%) and ribavirin (37.9%). Notably, the curative activity of compound 5i was up to 79.5%, with the EC50 value of 75.9 µg/mL, whereas the EC50 value of ningnanmycin was 362.4 µg/mL. The pot experiments also further demonstrated the significantly curative effect of 5i. Meanwhile, compounds 5h, 5p and 5x displayed more protective activities on TMV than that of ningnanmycin. Moreover, compounds 5a, 5e, 5f, and 5i showed inactivation activity similar to ningnanmycin at 500 µg/mL, and the EC50 value of 5e (41.5 µg/mL) was lower than ningnanmycin (50.0 µg/mL). The findings of transmission electron microscopic (TEM) indicated that the synthesized compounds exhibited strong and significant binding affinity to TMV coat protein (CP) and could obstruct the self-assembly and increment of TMV particles. Microscale thermophoresis (MST) studies on TMV-CP and CMV CP revealed that some of the active compounds, particularly 5i, exhibited a strong binding capability to TMV-CP or CMV-CP. This study revealed that anthranilic diamide derivatives containing moieties of trifluoromethylpyridine and hydrazone could be used as novel antiviral agents for controlling the plant viruses.


Assuntos
Antivirais/síntese química , Antivirais/farmacologia , Diamida/química , Hidrazonas/química , Vírus de Plantas/efeitos dos fármacos , Piridinas/química , Antivirais/química , Cucumovirus/efeitos dos fármacos , Cucumovirus/crescimento & desenvolvimento , Diamida/farmacologia , Desenho de Fármacos , Hidrazonas/farmacologia , Testes de Sensibilidade Microbiana , Vírus de Plantas/crescimento & desenvolvimento , Piridinas/síntese química , Piridinas/farmacologia , Relação Estrutura-Atividade , Vírus do Mosaico do Tabaco/efeitos dos fármacos , Vírus do Mosaico do Tabaco/crescimento & desenvolvimento
4.
J Agric Food Chem ; 67(8): 2148-2156, 2019 Feb 27.
Artigo em Inglês | MEDLINE | ID: mdl-30730738

RESUMO

Plant viral diseases seriously affect crop yield and quality. The natural product gramine (1) and its simple structural analogues 2-35 were synthesized from indoles, amines, and aldehydes in one step. The antiviral effects of these alkaloids were evaluated systematically. Most of these compounds were found to have higher antiviral effects than commercial ribavirin for the first time. Especially compounds 22, 30, and 31 exhibited significantly higher effects than ningnanmycin, thereby emerging as novel antiviral leads for further optimization. The preliminary implementation indicated that these compounds likely inhibit the assembly of tobacco mosaic virus (TMV) by cross-linking TMV capsid protein. Gramine analogues were also found to have broad-spectrum fungicidal effects. Although gramine has been reported to have influence on germination and development of Erysiphe graminis, these compounds displayed no fungicidal effects against Blumeria graminis f. sp. tritici on wheat in our test. Some of these compounds also exhibited certain insecticidal activities.


Assuntos
Antivirais/farmacologia , Produtos Biológicos/farmacologia , Alcaloides Indólicos/farmacologia , Antivirais/química , Produtos Biológicos/química , Descoberta de Drogas , Alcaloides Indólicos/química , Vírus de Plantas/efeitos dos fármacos , Vírus de Plantas/fisiologia , Relação Estrutura-Atividade , Vírus do Mosaico do Tabaco/efeitos dos fármacos , Vírus do Mosaico do Tabaco/fisiologia
5.
J Agric Food Chem ; 67(7): 1795-1806, 2019 Feb 20.
Artigo em Inglês | MEDLINE | ID: mdl-30681853

RESUMO

Plant viral diseases cause tremendous decreases in crop yield and quality. Natural products have always been a valuable source for lead discovery in medicinal and agricultural chemistry. A series of pimprinine alkaloids and their derivatives were prepared and identified by nuclear magnetic resonance (NMR) spectroscopy and high-resolution mass spectrometry (HR-MS). The antiviral activities of these alkaloids against tobacco mosaic virus (TMV) were systematically investigated for the first time. Most of the compounds exhibited higher antiviral activities than ribavirin. Compounds 5l, 9h, and 10h, which had similar or higher antiviral activities than ningnanmycin (perhaps the most widely used antiviral agent at present), emerged as new antiviral pilot compounds. This systematic structure-activity-relationship research lays the foundation for simplifying the structure of these alkaloids. The ring-open products, acylhydrazones 9a-9u, were also found to possess good antiviral activities. Moreover, all the synthesized compounds displayed broad-spectrum fungicidal activities. This study provides important information for the research and development of pimprinine alkaloids as novel antiviral agents.


Assuntos
Alcaloides/farmacologia , Antivirais , Oxazóis/farmacologia , Alcaloides/química , Citidina/análogos & derivados , Citidina/farmacologia , Estrutura Molecular , Oxazóis/química , Doenças das Plantas/virologia , Vírus de Plantas/efeitos dos fármacos , Ribavirina/farmacologia , Relação Estrutura-Atividade , Vírus do Mosaico do Tabaco/efeitos dos fármacos
6.
J Agric Food Chem ; 66(21): 5335-5345, 2018 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-29741370

RESUMO

A series of dithioacetal derivatives bearing a strobilurin moiety were designed and synthesized on the basis of our previous work. The antiviral activities of these compounds against Potato virus Y (PVY), Cucumber mosaic virus (CMV), and Tobacco mosaic virus (TMV) were systematically evaluated. Bioassay results indicated that C14 elicited excellent curative and protective activities against PVY, CMV, and TMV. The former had 50% effective concentrations (EC50) of 125.3, 108.9, and 181.7 µg/mL, respectively, and the latter had 148.4, 113.2, and 214.6 µg/mL, respectively, which were significantly superior to those of lead compound 6f (297.6, 259.6, and 582.4 µg/mL and 281.5, 244.3, and 546.3 µg/mL, respectively), Ningnanmycin (440.5, 549.1, and 373.8 µg/mL and 425.3, 513.3, and 242.7 µg/mL, respectively), Chitosan oligosaccharide (553.4, 582.8, and 513.8 µg/mL and 547.3, 570.6, and 507.9 µg/mL, respectively), and Ribavirin (677.4, 690.3, and 686.5 µg/mL and 652.7, 665.4, and 653.4 µg/mL, respectively). Moreover, defensive enzyme activities and RT-qPCR analysis demonstrated that the antiviral activity was associated with the changes of SOD, CAT, and POD activities in tobacco, which was proved by the related proteins of abscisic acid signaling pathway. This work provided a basis for further design, structural modification, and development of dithioacetal derivatives as new antiviral agents.


Assuntos
Acetais/química , Antivirais/farmacologia , Doenças das Plantas/virologia , Vírus de Plantas/efeitos dos fármacos , Estrobilurinas/farmacologia , Acetais/farmacologia , Antivirais/química , Clorofila/análise , Cucumovirus/efeitos dos fármacos , Doenças das Plantas/prevenção & controle , Folhas de Planta/virologia , Potyvirus/efeitos dos fármacos , Estrobilurinas/química , Nicotiana/virologia , Vírus do Mosaico do Tabaco/efeitos dos fármacos
7.
Plant Signal Behav ; 12(6): e1338223, 2017 06 03.
Artigo em Inglês | MEDLINE | ID: mdl-28594275

RESUMO

Reactive oxygen species (ROS), including superoxide anion (O2-), hydrogen peroxide (H2O2), and hydroxyl radical, act as signaling molecules to transduce biotic and abiotic stimuli into stress adaptations in plants. A respiratory burst oxidase homolog B of Nicotiana benthamiana (NbRBOHB) is responsible for O2- production to inhibit pathogen infection during plant innate immunity. RBOH-derived O2- can be immediately converted into H2O2 by the action of superoxide dismutase. Interestingly, we recently showed that red clover necrotic mosaic virus (RCNMV), a plant positive-strand RNA [(+)RNA] virus, hijacks the host's ROS-generating machinery during infection. An RCNMV replication protein associates with NbRBOHB and triggers intracellular ROS bursts. These bursts are required for robust viral RNA replication. However, what types of ROS are required for viral replication is currently unknown. Here, we found that RCNMV replication was sensitive to an O2- scavenger but insensitive to an H2O2 scavenger. Interestingly, replication of another plant (+)RNA virus, brome mosaic virus, was sensitive to both types of scavengers. These results indicate a virus-specific pattern requirement of O2- and H2O2 for (+)RNA virus replication and suggest a conserved nature of the roles of ROS in (+)RNA virus replication.


Assuntos
Peróxido de Hidrogênio/metabolismo , Nicotiana/virologia , Vírus de Plantas/fisiologia , Vírus de RNA/fisiologia , Superóxidos/metabolismo , Replicação Viral/fisiologia , Sequestradores de Radicais Livres/farmacologia , Vírus de Plantas/efeitos dos fármacos , Vírus de RNA/efeitos dos fármacos , Nicotiana/efeitos dos fármacos , Replicação Viral/efeitos dos fármacos
8.
Pestic Biochem Physiol ; 135: 15-26, 2017 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-28043326

RESUMO

Plant virus diseases, known as 'plant cancer', are the second largest plant diseases after plant fungal diseases, which have caused great damage to agricultural industry. Since now, the most direct and effective method for controlling viruses is chemotherapeutics, except for screening of anti-disease species. As the occurrence and harm of plant diseases intensify, production and consumption of pesticides have increased year by year, and greatly contributed to the fertility of agriculture, but also brought a series of problems, such as the increase of drug resistance of plant pathogens and the excessive pesticide residues. In recent years, biopesticide, as characterized by environmentally safe due to low residual, safe to non-target organism due to better specificity and not as susceptible to produce drug resistance due to diverse work ways, has gained more attention than ever before and exhibited great development potential. Now much progress has been made about researches on new biogenic anti-plant-virus substances. The types of active components include proteins, polysaccharides and small molecules (alkaloids, flavonoids, phenols, essential oils) from plants, proteins and polysaccharides from microorganisms, polysaccharides from algae and oligochitosan from animals. This study summarized the research advance of biogenic anti-plant-virus substances in recent years and put forward their further development in the future.


Assuntos
Antivirais/farmacologia , Vírus de Plantas/efeitos dos fármacos , Animais , Bactérias/química , Fungos/química , Compostos Fitoquímicos/farmacologia , Preparações de Plantas/química , Proteínas de Plantas , Plantas/química
9.
Nat Plants ; 3: 16207, 2017 01 09.
Artigo em Inglês | MEDLINE | ID: mdl-28067898

RESUMO

Topical application of pathogen-specific double-stranded RNA (dsRNA) for virus resistance in plants represents an attractive alternative to transgenic RNA interference (RNAi). However, the instability of naked dsRNA sprayed on plants has been a major challenge towards its practical application. We demonstrate that dsRNA can be loaded on designer, non-toxic, degradable, layered double hydroxide (LDH) clay nanosheets. Once loaded on LDH, the dsRNA does not wash off, shows sustained release and can be detected on sprayed leaves even 30 days after application. We provide evidence for the degradation of LDH, dsRNA uptake in plant cells and silencing of homologous RNA on topical application. Significantly, a single spray of dsRNA loaded on LDH (BioClay) afforded virus protection for at least 20 days when challenged on sprayed and newly emerged unsprayed leaves. This innovation translates nanotechnology developed for delivery of RNAi for human therapeutics to use in crop protection as an environmentally sustainable and easy to adopt topical spray.


Assuntos
Silicatos de Alumínio/farmacologia , Nanoestruturas/química , Doenças das Plantas/prevenção & controle , Vírus de Plantas/efeitos dos fármacos , Interferência de RNA , RNA de Cadeia Dupla/farmacologia , RNA Viral/farmacologia , Arabidopsis/fisiologia , Argila , Doenças das Plantas/virologia , Vírus de Plantas/genética , Nicotiana/fisiologia , Vigna/fisiologia
10.
PLoS One ; 10(2): e0117496, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25705894

RESUMO

A novel acetone extract from cottonseed oil sludge was firstly discovered against plant viruses including Tobacco mosaic virus (TMV), Rice stripe virus (RSV) and Southern rice black streaked dwarf virus (SRBSDV). Gossypol and ß-sitosterol separated from the acetone extract were tested for their effects on anti-TMV and analysed by nuclear magnetic resonance (NMR) assay. In vivo and field trials in different geographic distributions and different host varieties declared that this extract mixture was more efficient than the commercial agent Ningnanmycin with a broad spectrum of anti-plant-viruses activity. No phytotoxic activity was observed in the treated plants and environmental toxicology showed that this new acetone extract was environmentally friendly, indicating that this acetone extract has potential application in the control of plant virus in the future.


Assuntos
Acetona/química , Antivirais/farmacologia , Óleo de Sementes de Algodão/química , Extratos Vegetais/farmacologia , Vírus de Plantas/efeitos dos fármacos
11.
Virol J ; 12: 5, 2015 Jan 27.
Artigo em Inglês | MEDLINE | ID: mdl-25623384

RESUMO

BACKGROUND: In recent years, a number of serious disease outbreaks caused by viruses and viroids on greenhouse tomatoes in North America have resulted in significant economic losses to growers. The objectives of this study were to evaluate the effectiveness of commercial disinfectants against mechanical transmission of these pathogens, and to select disinfectants with broad spectrum reactivity to control general virus and viroid diseases in greenhouse tomato production. METHODS: A total of 16 disinfectants were evaluated against Pepino mosaic virus (PepMV), Potato spindle tuber viroid (PSTVd), Tomato mosaic virus (ToMV), and Tobacco mosaic virus (TMV). The efficacy of each disinfectant to deactivate the pathogen's infectivity was evaluated in replicate experiments from at least three independent experiments. Any infectivity that remained in the treated solutions was assessed through bioassays on susceptible tomato plants through mechanical inoculation using inocula that had been exposed with the individual disinfectant for three short time periods (0-10 sec, 30 sec and 60 sec). A positive infection on the inoculated plant was determined through symptom observation and confirmed with enzyme-linked immunosorbent assay (PepMV, ToMV, and TMV) and real-time reverse transcription-PCR (PSTVd). Experimental data were analyzed using Logistic regression and the Bayesian methodology. RESULTS: Statistical analyses using logistic regression and the Bayesian methodology indicated that two disinfectants (2% Virkon S and 10% Clorox regular bleach) were the most effective to prevent transmission of PepMV, PSTVd, ToMV, and TMV from mechanical inoculation. Lysol all-purpose cleaner (50%) and nonfat dry milk (20%) were also effective against ToMV and TMV, but with only partial effects for PepMV and PSTVd. CONCLUSION: With the broad spectrum efficacy against three common viruses and a viroid, several disinfectants, including 2% Virkon S, 10% Clorox regular bleach and 20% nonfat dry milk, are recommend to greenhouse facilities for consideration to prevent general virus and viroid infection on tomato plants.


Assuntos
Desinfetantes/farmacologia , Doenças das Plantas/prevenção & controle , Doenças das Plantas/virologia , Vírus de Plantas/efeitos dos fármacos , Solanum lycopersicum/crescimento & desenvolvimento , Solanum lycopersicum/virologia , Viroides/efeitos dos fármacos , Antígenos Virais/análise , Bioensaio , Ensaio de Imunoadsorção Enzimática , Viabilidade Microbiana/efeitos dos fármacos , Vírus de Plantas/isolamento & purificação , Inativação de Vírus
13.
J Agric Food Chem ; 61(43): 10302-9, 2013 Oct 30.
Artigo em Inglês | MEDLINE | ID: mdl-24134653

RESUMO

An indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) was developed for the quantitative detection of the antiviral agent against tobacco mosaic virus (TMV). Hapten 6-[2-((methyllbenzothiazol-2-yl)-1-(2-ethoxy)-O, O-diethyl-α-aminophosphonate)acetamido)] hexanoic acid (DHS) was prepared from commercial chemicals and incorporated into the spacer arm through a carbon-carbon single bond. The prepared hapten was then coupled to carrier proteins keyhole limpet hemocyanin (KLH) to be used as an immunogen for monoclonal antibody (mAb) production together with ELISA development. This assay was further optimized by the assessment of the dependence of assay parameters on organic solvents, pH, and ionic strength. The IC50 values of the optimized assay for Dufulin and the calculated limit of detection in phosphate-buffered saline (PBS) were 9.6 ± 0.59 and 0.3 ± 0.05 ng/mL, respectively. Using the optimized assays Dufulin residues in soil and tobacco samples were determined with recovery values ranging from 81.5 to 95.3%, intra-assay variation ranging from 2.88 to 6.10%, and interassay variation ranging from 6.11 to 9.42%.


Assuntos
Antivirais/análise , Benzotiazóis/análise , Ensaio de Imunoadsorção Enzimática/métodos , Haptenos/química , Nicotiana/química , Poluentes do Solo/análise , Anticorpos Monoclonais/análise , Resíduos de Drogas/análise , Ensaio de Imunoadsorção Enzimática/instrumentação , Doenças das Plantas/virologia , Vírus de Plantas/efeitos dos fármacos , Vírus de Plantas/fisiologia , Nicotiana/virologia
14.
J Agric Food Chem ; 61(34): 8049-55, 2013 Aug 28.
Artigo em Inglês | MEDLINE | ID: mdl-23915352

RESUMO

Southern rice black-streaked dwarf virus (SRBSDV) is a rice pathogen that had an outbreak in southern China in 2010 and caused significant crop losses. Therefore, screening for effective antiviral drugs against SRBSDV is very important. This study used rice suspension cells infected with SRBSDV by polyethylene glycol-mediated uptake for screening antiviral drugs. SRBSDV P7-1, which is coded by the S7-1 gene, has an intrinsic ability to self-interact to form tubules that play an important role in viral infection. Therefore, relative expression level of the SRBSDV S7-1 gene in infected rice suspension cells was assayed by real-time quantitative polymerase chain reaction to evaluate the antiviral activities of various drugs. Dufulin displayed the highest inhibitory activity against SRBSDV S7-1 expression. In addition, changes in peroxidase (POD), polyphenol oxidase (PPO), and phenylalanine ammonia-lyase (PAL) activities were determined in inoculated and noninoculated cells. The results showed that both POD and PPO activities increased upon dufulin treatment. Furthermore, the validity of this approach was confirmed in an in vivo experiment in which dufulin was found to effectively inhibit SRBSDV.


Assuntos
Antivirais/farmacologia , Oryza/virologia , Doenças das Plantas/virologia , Vírus de Plantas/efeitos dos fármacos , Reoviridae/efeitos dos fármacos , Proteínas Virais/antagonistas & inibidores , Proteínas Virais/genética , Benzotiazóis/farmacologia , Técnicas de Cultura de Células , Avaliação Pré-Clínica de Medicamentos , Regulação Viral da Expressão Gênica/efeitos dos fármacos , Vírus de Plantas/genética , Vírus de Plantas/metabolismo , Reoviridae/genética , Reoviridae/metabolismo , Proteínas Virais/metabolismo
15.
Plant Mol Biol ; 81(4-5): 507-22, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23400832

RESUMO

The phytotoxic effects of copper (Cu) and cadmium (Cd) on plant growth are well documented. However, Cu and Cd toxicity targets and the cellular systems contributing to acquisition of tolerance are not fully understood at the molecular level. We aimed to identify genes and pathways that discriminate the actions of Cu and Cd in rice roots (Oryza sativa L. cv. TN67). The transcripts of 1,450 and 1,172 genes were regulated after Cu and Cd treatments, respectively. We identified 882 genes specifically respond to Cu treatment, and 604 unique genes as Cd-responsive by comparison of expression profiles of these two regulated gene groups. Gene ontology analysis for 538 genes involved in primary metabolism, oxidation reduction and response to stimulus was changed in response to both metals. In the individual aspect, Cu specifically altered levels of genes involved in vesicle trafficking transport, fatty acid metabolism and cellular component biogenesis. Cd-regulated genes related to unfolded protein binding and sulfate assimilation. To further characterize the functions of vesicle trafficking transport under Cu stress, interference of excytosis in root tissues was conducted by inhibitors and silencing of Exo70 genes. It was demonstrated that vesicle-trafficking is required for mediation of Cu-induced reactive oxygen species (ROS) production in root tissues. These results may provide new insights into understanding the molecular basis of the early metal stress response in plants.


Assuntos
Cádmio/toxicidade , Cobre/toxicidade , Oryza/efeitos dos fármacos , Oryza/genética , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/genética , Transcriptoma/genética , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Inativação Gênica/efeitos dos fármacos , Genes de Plantas/genética , Anotação de Sequência Molecular , Análise de Sequência com Séries de Oligonucleotídeos , Oryza/crescimento & desenvolvimento , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Raízes de Plantas/crescimento & desenvolvimento , Vírus de Plantas/efeitos dos fármacos , Vírus de Plantas/genética , Espécies Reativas de Oxigênio/metabolismo , Reprodutibilidade dos Testes , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Plântula/efeitos dos fármacos , Plântula/genética , Plântula/crescimento & desenvolvimento , Estresse Fisiológico/efeitos dos fármacos , Estresse Fisiológico/genética , Nicotiana/efeitos dos fármacos , Nicotiana/genética , Nicotiana/virologia , Transcriptoma/efeitos dos fármacos , Vesículas Transportadoras/efeitos dos fármacos , Vesículas Transportadoras/metabolismo
16.
Planta ; 237(1): 77-88, 2013 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-22983699

RESUMO

Alpha-momorcharin (α-MMC) is type-1 ribosome inactivating proteins (RIPs) with molecular weight of 29 kDa and has lots of biological activity. Our recent study indicated that the α-MMC purified from seeds of Momordica charantia exhibited distinct antiviral and antifungal activity. Tobacco plants pre-treated with 0.5 mg/mL α-MMC 3 days before inoculation with various viruses showed less-severe symptom and less reactive oxygen species (ROS) accumulation compared to that inoculated with viruses only. Quantitative real-time PCR analysis revealed that the replication levels of viruses were lower in the plants treated with the α-MMC than control plants at 15 days post inoculation. Moreover, the coat protein expression of viruses was almost completely inhibited in plants which were treated with the α-MMC compared with control plants. Furthermore, the SA-responsive defense-related genes including non-expressor of pathogenesis-related genes 1 (NPR1), PR1, PR2 were up-regulated and activities of some antioxidant enzymes including superoxide dismutase (SOD), catalase (CAT), peroxidase (POD) were increased after the α-MMC treatment. In addition, the α-MMC (500 µg/mL) revealed remarkable antifungal effect against phytopathogenic fungi, in the growth inhibition range 50.35-67.21 %, along with their MIC values ranging from 100 to 500 µg/mL. The α-MMC had also a strong detrimental effect on spore germination of all the tested plant pathogens along with concentration as well as time-dependent kinetic inhibition of Sclerotinia sclerotiorum. The α-MMC showed a remarkable antiviral and antifungal effect and hence could possibly be exploited in crop protection for controlling certain important plant diseases.


Assuntos
Antifúngicos/farmacologia , Resistência à Doença/efeitos dos fármacos , Nicotiana/metabolismo , Vírus de Plantas/efeitos dos fármacos , Proteínas Inativadoras de Ribossomos/farmacologia , Ascomicetos/efeitos dos fármacos , Ascomicetos/crescimento & desenvolvimento , Aspergillus niger/efeitos dos fármacos , Aspergillus niger/crescimento & desenvolvimento , Aspergillus oryzae/efeitos dos fármacos , Aspergillus oryzae/crescimento & desenvolvimento , Western Blotting , Proteínas do Capsídeo/genética , Catalase/metabolismo , Resistência à Doença/genética , Fusarium/efeitos dos fármacos , Fusarium/crescimento & desenvolvimento , Regulação da Expressão Gênica de Plantas/genética , Regulação Viral da Expressão Gênica/efeitos dos fármacos , Interações Hospedeiro-Patógeno/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Momordica charantia/química , Peroxidase/metabolismo , Doenças das Plantas/genética , Doenças das Plantas/virologia , Folhas de Planta/genética , Folhas de Planta/metabolismo , Folhas de Planta/virologia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Vírus de Plantas/genética , Vírus de Plantas/fisiologia , Espécies Reativas de Oxigênio/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Superóxido Dismutase/metabolismo , Nicotiana/genética , Nicotiana/virologia , Replicação Viral/efeitos dos fármacos , Replicação Viral/genética
17.
Plant Cell Rep ; 30(8): 1427-34, 2011 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-21404008

RESUMO

Effect of various lead (Pb) concentrations on the systemic movement of RNA viruses was examined in tobacco plants. Prior to inoculation, plants were grown hydroponically for 6 days in Hoagland's solution supplemented with five concentrations of lead nitrate [Pb(NO(3))(2)]: 0.0 (control), 10, 15, 50, and 100 µM. Four different RNA viruses with different cell-to-cell movement mechanisms were used. Two weeks after inoculation lower and upper leaves of each treatment were harvested and examined for the presence of viral coat protein. In plants inoculated with Tobacco mosaic virus, Potato virus X, and Tobacco etch virus, TEM images and western blot assays confirmed the presence of viral coat proteins in the upper leaves of all lead treatments. However, in plants inoculated with Turnip vein-clearing virus (TVCV), no signs of viral particles were detected in the upper leaves of plants treated with 10 µM or 15 µM lead nitrate. In contrast, plants treated with high concentrations of lead nitrate (50 µM or 100 µM) showed viral particles in their upper leaves. In plants treated with 10 µM or 15 µM lead nitrate, callose accumulation was the same as in control plants. This suggests that non-toxic concentrations of lead nitrate may trigger the production of putative cellular factors in addition to callose that interfere with the TVCV systemic movement. In contrast, plants treated with 100 µM lead nitrate showed less callose as compared to control plants, facilitating the systemic movement of TVCV.


Assuntos
Chumbo/farmacologia , Nicotiana/virologia , Vírus de Plantas/efeitos dos fármacos , Vírus de RNA/efeitos dos fármacos , Proteínas do Capsídeo/isolamento & purificação , Glucanos/análise , Microscopia Eletrônica de Transmissão , Nitratos/farmacologia , Folhas de Planta/ultraestrutura , Folhas de Planta/virologia , Vírus de Plantas/fisiologia , Vírus de RNA/fisiologia
18.
Mol Plant Microbe Interact ; 24(6): 706-18, 2011 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-21281112

RESUMO

Tomato ringspot virus (ToRSV, a Nepovirus sp.) systemically infects many herbaceous plants. Viral RNA accumulates in symptomatic leaves and in young, asymptomatic leaves that emerge late in infection. Here, we show that systemic infection by ToRSV is restricted in tobacco. After an initial hypersensitive response in inoculated leaves, only a few plants showed limited systemic symptoms. Viral RNA did not usually accumulate to detectable levels in asymptomatic leaves. ToRSV-derived small-interfering RNAs and PR1a transcripts were only detected in tissues that contained viral RNA, indicating local induction of RNA silencing and salicylic acid (SA)-dependent defense responses. Lesion size and viral systemic spread were reduced with SA pretreatment but enhanced in NahG transgenic lines deficient in SA accumulation, suggesting that SA-dependent mechanisms play a key role in limiting ToRSV spread in tobacco. Restriction of virus infection was enhanced in transgenic lines expressing the P1-HC-Pro suppressor of silencing. Knocking down the SA-inducible RNA-dependent RNA polymerase 1 exacerbated the necrotic reaction but did not affect viral systemic spread. ToRSV-infected tobacco plants were susceptible to reinoculation by ToRSV or Tobacco mosaic virus, although a small reduction in lesion size was observed. This moderate systemic resistance suggests inefficient induction or spread of RNA silencing and systemic acquired resistance signal molecules.


Assuntos
Nicotiana/virologia , Doenças das Plantas/virologia , Vírus de Plantas/efeitos dos fármacos , Vírus de Plantas/fisiologia , Interferência de RNA , Ácido Salicílico/metabolismo , Regulação da Expressão Gênica de Plantas/fisiologia , Interações Hospedeiro-Patógeno , Folhas de Planta/virologia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , RNA Interferente Pequeno , Transdução de Sinais , Temperatura
19.
Planta ; 233(2): 299-308, 2011 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-21046144

RESUMO

Plant viruses cause many diseases that lead to significant economic losses. However, most of the approaches to control plant viruses, including transgenic processes or drugs are plant-species-limited or virus-species-limited, and not very effective. We introduce an application of jasmonic acid (JA) and salicylic acid (SA), a broad-spectrum, efficient and nontransgenic method, to improve plant resistance to RNA viruses. Applying 0.06 mM JA and then 0.1 mM SA 24 h later, enhanced resistance to Cucumber mosaic virus (CMV), Tobacco mosaic virus (TMV) and Turnip crinkle virus (TCV) in Arabidopsis, tobacco, tomato and hot pepper. The inhibition efficiency to virus replication usually achieved up to 80-90%. The putative molecular mechanism was investigated. Some possible factors affecting the synergism of JA and SA have been defined, including WRKY53, WRKY70, PDF1.2, MPK4, MPK2, MPK3, MPK5, MPK12, MPK14, MKK1, MKK2, and MKK6. All genes involving in the synergism of JA and SA were investigated. This approach is safe to human beings and environmentally friendly and shows potential as a strong tool for crop protection against plant viruses.


Assuntos
Arabidopsis/virologia , Ciclopentanos/farmacologia , Oxilipinas/farmacologia , Doenças das Plantas/virologia , Vírus de Plantas/efeitos dos fármacos , Ácido Salicílico/farmacologia , Solanaceae/virologia , Antivirais/administração & dosagem , Antivirais/farmacologia , Ciclopentanos/administração & dosagem , Oxilipinas/administração & dosagem , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/virologia , Ácido Salicílico/administração & dosagem , Replicação Viral/efeitos dos fármacos
20.
Amino Acids ; 33(2): 267-72, 2007 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-17429570

RESUMO

Polyamines, oxidized by serum amine oxidase, yield aminoaldehydes and hydrogen peroxide. Acrolein may be formed from the aminoaldehydes by a spontaneous beta-elimination process. These oxidation products "oxidized polyamines" inhibit bacterial growth and exhibit anticancer activity. The antimicrobial activity of oxidized polyamines is not limited to bacteria; and the inactivation of bacterial viruses, plant viruses and animal viruses, was also reported. Bacteriophages of the T-odd series are permeable and were inactivated by oxidized polyamines. The inactive phages absorb to their bacterial host and injected their DNA, which formed a stable inactive complex with the aminoaldehydes. Aminoaldehydes, synthesized chemically, also inactivated viruses. The growth of the plant viruses: Tobacco mosaic virus, Potato virus X and Alfalfa mosaic virus was also inhibited by oxidized polyamines. The animal viruses, which were inactivated by oxidized polyamines included Myxoviruses (influenza and Newcastle disease viruses), West Nile, vaccinia and Sindbis viruses. These findings may have practical implications.


Assuntos
Amina Oxidase (contendo Cobre)/metabolismo , Antivirais/farmacologia , Poliaminas/farmacologia , Acroleína/farmacologia , Aldeídos/farmacologia , Animais , Colífagos/efeitos dos fármacos , DNA Viral/metabolismo , Humanos , Peróxido de Hidrogênio/farmacologia , Vírus da Doença de Newcastle/efeitos dos fármacos , Orthomyxoviridae/efeitos dos fármacos , Oxirredução , Vírus de Plantas/efeitos dos fármacos , Sindbis virus/efeitos dos fármacos , Vírus do Nilo Ocidental/efeitos dos fármacos
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