RESUMO
BACKGROUND: The addictive potential of areca nut (AN) and tobacco is well-documented, but their combined neurobiological effects in AN-containing tobacco products (ANTP) remain obscure. This study employed a three-stage in silico approach to investigate the potential targets and pathways associated with the addictive properties of AN alone and in ANTP. MATERIALS AND METHODS: Bioactive molecules were retrieved for AN and tobacco, followed by target prediction and pathway enrichment analysis. The identified biomolecules were categorized into AN and ANTP groups. RESULTS: A total of 195 bioactive molecules were identifid (38 to AN, 157 to tobacco). Absorption, distribution, metabolism, and excretion (ADME) details were retrieved. Predicted bioactivity (gene/protein interaction probability ≥80%) was analysed, revealing 13 shared targets between AN and tobacco, 12 exclusive to AN, and 33 exclusive to tobacco. AN and ANTP influenced 21 and 27 pathways (FDR ≤ 0.05), respectively, with distinct footprints. Notably, GABAergic and cholinergic synapses, nicotine addiction, calcium signaling, and morphine addiction pathways were differentially enriched between AN and ANTP. DISCUSSION: This study highlights the distinct and synergistic neurobiological effects of AN and tobacco in ANTP. The identified differences in target genes and pathways underscore the need for tailored interventions and cessation strategies for users of AN and ANTP products. Further research is warranted to validate these findings, explore interplay between diverse addiction factors, and develop effective prevention and treatment programs. (Neuropsychopharmacol Hung 2024; 26(3): 133-143)
Assuntos
Areca , Comportamento Aditivo , Simulação por Computador , Nicotiana , Humanos , Comportamento Aditivo/psicologia , Comportamento Aditivo/metabolismo , Nozes , Tabagismo/psicologia , Tabagismo/metabolismo , Produtos do Tabaco/efeitos adversosRESUMO
BACKGROUND: Tobacco smoking is the leading cause of preventable death and disease worldwide, with over 8 million annual deaths attributed to cigarette smoking. This study investigates the impact of cigarette smoke and heated tobacco products (HTPs) on microglial function, focusing on toxicological profiles, inflammatory responses, and oxidative stress using ISO standard and clinically relevant conditions of exposure. METHODS: We assessed cell viability, reactive oxygen species (ROS) production, lipid peroxidation, mitochondrial function, unfolded protein response, and inflammation in human microglial cells (HMC3) exposed to cigarette smoke, HTP aerosol or nicotine. RESULTS: Our findings show that cigarette smoke significantly reduces microglial viability, increases ROS formation, induces lipid peroxidation, and reduces intracellular glutathione levels. Cigarette smoke also alters the expression of genes involved in mitochondrial dynamics and biogenesis, leading to mitochondrial dysfunction. Additionally, cigarette smoke impairs the unfolded protein response, activates the NF-κB pathway, and induces a pro-inflammatory state characterized by increased TNF and IL-18 expression. Furthermore, cigarette smoke causes DNA damage and decreases the expression of the aging marker Klotho ß. In contrast, HTP, exhibited a lesser degree of microglial toxicity, with reduced ROS production, lipid peroxidation, and mitochondrial dysfunction compared to conventional cigarettes. CONCLUSION: These results highlight the differential toxicological profile of cigarette smoke and HTP on microglial cells, suggesting a potential harm reduction strategy for neurodegenerative disease for smokers unwilling or unable to quit.
Assuntos
Sobrevivência Celular , Inflamação , Peroxidação de Lipídeos , Microglia , Mitocôndrias , Estresse Oxidativo , Espécies Reativas de Oxigênio , Fumaça , Produtos do Tabaco , Resposta a Proteínas não Dobradas , Estresse Oxidativo/efeitos dos fármacos , Humanos , Espécies Reativas de Oxigênio/metabolismo , Inflamação/patologia , Microglia/efeitos dos fármacos , Microglia/metabolismo , Microglia/patologia , Produtos do Tabaco/efeitos adversos , Fumaça/efeitos adversos , Mitocôndrias/metabolismo , Mitocôndrias/efeitos dos fármacos , Peroxidação de Lipídeos/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Resposta a Proteínas não Dobradas/efeitos dos fármacos , Linhagem Celular , Temperatura Alta , NF-kappa B/metabolismo , Nicotiana/efeitos adversos , Dano ao DNARESUMO
Autophagy, an intracellular degradation process, has emerged as a crucial innate immune response against various plant pathogens, including viruses. Tomato spotted wilt orthotospovirus (TSWV) is a highly destructive plant pathogen that infects over 1000 plant species and poses a significant threat to global food security. However, the role of autophagy in defence against the TSWV pathogen, and whether the virus counteracts this defence, remains unknown. In this study, we report that autophagy plays an important role in antiviral defence against TSWV infection; however, this autophagy-mediated defence is counteracted by the viral effector NSs. Transcriptome profiling revealed the up-regulation of autophagy-related genes (ATGs) upon TSWV infection. Blocking autophagy induction by chemical treatment or knockout/down of ATG5/ATG7 significantly enhanced TSWV accumulation. Notably, the TSWV nucleocapsid (N) protein, a major component of the viral replication unit, strongly induced autophagy. However, the TSWV nonstructural protein NSs was able to effectively suppress N-induced autophagy in a dose-dependent manner. Further investigation revealed that NSs inhibited ATG6-mediated autophagy induction. These findings provide new insights into the defence role of autophagy against TSWV, a representative segmented negative-strand RNA virus, as well as the tospoviral pathogen counterdefence mechanism.
Assuntos
Autofagia , Doenças das Plantas , Tospovirus , Tospovirus/fisiologia , Tospovirus/patogenicidade , Doenças das Plantas/virologia , Doenças das Plantas/imunologia , Proteínas não Estruturais Virais/metabolismo , Proteínas não Estruturais Virais/genética , Solanum lycopersicum/virologia , Solanum lycopersicum/imunologia , Solanum lycopersicum/genética , Nicotiana/virologia , Nicotiana/imunologia , Nicotiana/genéticaRESUMO
Tobacco mosaic virus (TMV) is extremely pathogenic and resistant to stress There are great needs to develop methods to reduce the virus in the environment and induce plant immunity simultaneously. Here, we report a multifunctional nano-protectant to reduce the virus in the environment and induce plant immunity simultaneously. The star polycation (SPc) nanocarrier can act as an active ingredient to interact with virus coat protein via electrostatic interaction, which reduces the proportion of TMV particles to 2.9% and leads to a reduction of the amount of virus in the environment by half. SPc can act as an adjuvant to spontaneously assemble with an immune inducer lentinan (LNT) through hydrogen bonding into nanoscale (142 nm diameter) LNT/SPc complex, which improves the physicochemical property of LNT for better wetting performance on leaves and cellular uptake, and further activates plant immune responses. Finally, the LNT/SPc complex displays preventive and curative effects on TMV disease, reducing TMV-GFP relative expression by 26% in the laboratory and achieving 82% control efficacy in the field We hope the strategy reported here would be useful for control of crop virus disease.
Assuntos
Nicotiana , Doenças das Plantas , Imunidade Vegetal , Vírus do Mosaico do Tabaco , Vírus do Mosaico do Tabaco/efeitos dos fármacos , Vírus do Mosaico do Tabaco/imunologia , Doenças das Plantas/virologia , Doenças das Plantas/prevenção & controle , Doenças das Plantas/imunologia , Imunidade Vegetal/efeitos dos fármacos , Nicotiana/virologia , Nicotiana/imunologia , Nanoestruturas/química , Lentinano/farmacologia , Folhas de Planta/virologia , Proteínas do Capsídeo/imunologia , Proteínas do Capsídeo/metabolismoRESUMO
Although deterioration of silicone maxillofacial prostheses is severely accentuated in smoking patients, the phenomenon has not been systematically studied. To address a gap in the literature concerning the stability of maxillofacial prostheses during service, in this contribution, the effect of cigarette smoke on the aspect and physical properties of M511 silicone elastomer was evaluated. The aspect, surface, and overall properties of the silicone material, pigmented or not, were followed by AFM, color measurements, FTIR, water contact angle measurements, TGA-DTG and DSC, hardness and compression stress-strain measurements. The types of the contaminants adsorbed were assessed by XRF, ESI-MS, MALDI-MS, and NMR spectral analyses. Important modifications in color, contact angle, surface roughness, local mechanical properties, and thermal properties were found in the silicone material for maxillofacial prostheses after exposure to cigarettes smoke. The presence of lead, nicotine, and several other organic compounds adsorbed into the silicone material was emphasized. Slight decrease in hardness and increase in Young's modulus was found. The combined data show important impact of cigarette smoke on the silicone physical properties and could indicate chemical transformations by secondary cross-linking. To our knowledge, this is the first study making use of complementary physical methods to assess the effect of cigarette smoke on the aspect and integrity of silicone materials for maxillofacial prostheses.
Assuntos
Teste de Materiais , Prótese Maxilofacial , Fumaça , Humanos , Elastômeros de Silicone/química , Nicotiana/química , CorRESUMO
Pathogenesis-related proteins (PR), including osmotins, play a vital role in plant defense, being activated in response to diverse biotic and abiotic stresses. Despite their significance, the mechanistic insights into the role of osmotins in plant defense have not been extensively explored. The present study explores the cloning and characterization of the osmotin gene (WsOsm) from Withania somnifera, aiming to illuminate its role in plant defense mechanisms. Quantitative real-time PCR analysis revealed significant induction of WsOsm in response to various phytohormones e.g. abscisic acid, salicylic acid, methyl jasmonate, brassinosteroids, and ethrel, as well as biotic and abiotic stresses like heat, cold, salt, and drought. To further elucidate WsOsm's functional role, we overexpressed the gene in Nicotiana tabacum, resulting in heightened resistance against the Alternaria solani pathogen. Additionally, we observed enhancements in shoot length, root length, and root biomass in the transgenic tobacco plants compared to wild plants. Notably, the WsOsm- overexpressing seedlings demonstrated improved salt and drought stress tolerance, particularly at the seedling stage. Confocal histological analysis of H2O2 and biochemical studies of antioxidant enzyme activities revealed higher levels in the WsOsm overexpressing lines, indicating enhanced antioxidant defense. Furthermore, a pull-down assay and mass spectrometry analysis revealed a potential interaction between WsOsm and defensin, a known antifungal PR protein (WsDF). This suggests a novel role of WsOsm in mediating plant defense responses by interacting with other PR proteins. Overall, these findings pave the way for potential future applications of WsOsm in developing stress-tolerant crops and improving plant defense strategies against pathogens.
Assuntos
Defensinas , Regulação da Expressão Gênica de Plantas , Nicotiana , Proteínas de Plantas , Plantas Geneticamente Modificadas , Estresse Fisiológico , Withania , Withania/genética , Withania/fisiologia , Withania/metabolismo , Withania/efeitos dos fármacos , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Nicotiana/genética , Nicotiana/fisiologia , Nicotiana/efeitos dos fármacos , Nicotiana/microbiologia , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Estresse Fisiológico/genética , Defensinas/genética , Defensinas/metabolismo , Reguladores de Crescimento de Plantas/metabolismo , Alternaria/fisiologia , Secas , Plântula/genética , Plântula/fisiologia , Plântula/efeitos dos fármacos , Ácido Salicílico/metabolismo , Doenças das Plantas/microbiologia , Doenças das Plantas/genética , Doenças das Plantas/imunologia , Peróxido de Hidrogênio/metabolismo , Ácido Abscísico/metabolismo , Ácido Abscísico/farmacologia , Raízes de Plantas/genética , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/fisiologiaRESUMO
The leaf apoplast contains several compounds that play important roles in the regulation of different physiological processes in plants. However, this compartment has been neglected in several experimental and modelling studies, which is mostly associated to the difficulty to collect apoplast washing fluid (AWF) in sufficient amount for metabolomics analysis and as free as possible from symplastic contamination. Here, we established an approach based in an infiltration-centrifugation technique that use little leaf material but allows sufficient AWF collection for gas chromatography mass spectrometry (GC-MS)-based metabolomics analysis in both tobacco and Arabidopsis. Up to 54 metabolites were annotated in leaf and apoplast samples from both species using either 20% (v/v) methanol (20% MeOH) or distilled deionized water (ddH2O) as infiltration fluids. The use of 20% MeOH increased the yield of the AWF collected but also the level of symplastic contamination, especially in Arabidopsis. We propose a correction factor and recommend the use of multiple markers such as MDH activity, protein content and conductivity measurements to verify the level of symplastic contamination in MeOH-based protocols. Neither the concentration of sugars nor the level of primary metabolites differed between apoplast samples extracted with ddH2O or 20% MeOH. This indicates that ddH2O can be preferentially used, given that it is a non-toxic and highly accessible infiltration fluid. The infiltration-centrifugation-based approach established here uses little leaf material and ddH2O as infiltration fluid, being suitable for GC-MS-based metabolomics analysis in tobacco and Arabidopsis, with great possibility to be extended for other plant species and tissues.
Assuntos
Arabidopsis , Cromatografia Gasosa-Espectrometria de Massas , Metaboloma , Metabolômica , Nicotiana , Folhas de Planta , Folhas de Planta/metabolismo , Arabidopsis/metabolismo , Metabolômica/métodos , Nicotiana/metabolismo , Cromatografia Gasosa-Espectrometria de Massas/métodosRESUMO
Citrus huanglongbing (HLB), which is caused by the phloem-colonizing bacteria Candidatus Liberibacter asiaticus (CLas), poses a significant threat to citrus production worldwide. The pathogenicity mechanism of HLB remains poorly understood. SEC-dependent effectors (SDEs) have been suggested to play critical roles in the interaction between citrus and CLas. Here, we explored the function of CLIBASIA_05320 (SDE19), a core SDE from CLas, and its interaction with its host target. Our data revealed that SDE19 is expressed at higher level during infection of citrus than that during infection of the Asian citrus psyllid. Subcellular localization assays showed that SDE19 is localized in the nucleus and cytoplasm and is capable of moving from cell to cell in Nicotiana benthamiana. To investigate whether SDE19 facilitates pathogen infection, we generated transgenic Arabidopsis thaliana and citrus plants overexpressing SDE19. Transgenic A. thaliana and citrus plants were more susceptible to Pseudomonas syringae pv. tomato (Pst) and Xanthomonas citri subsp. citri (Xcc), respectively. In addition, RNA-seq analysis demonstrated that overexpression of SDE19 resulted in a reprogramming of expression of genes related to biotic stimulus responses. SDE19 interacts with Citrus sinensis Sec12, a guanine nucleotide exchange factor responsible for the assembly of plant COPII (coat protein II)-coated vesicles, which mediate vesicle trafficking from the ER to the Golgi. SDE19 colocalizes with Sec12 in the ER by binding to its N-terminal catalytic region, affecting the stability of Sec12 through the 26S proteasome. This interaction hinders the secretion of apoplastic defense-related proteins such as PR1, P69B, GmGIP1, and RCR3. Furthermore, the secretion of PR1 and callose deposition is decreased in SDE19-transgenic A. thaliana. Taken together, SDE19 is a novel virulent SDE secreted by CLas that interacts with Sec12 to disrupt vesicle trafficking, inhibit defense-related proteins secretion, and promote bacterial infection. This study sheds light on how CLas manipulates the host vesicle trafficking pathway to suppress the secretion of defense-related proteins and interfere with plant immunity.
Assuntos
Citrus sinensis , Doenças das Plantas , Imunidade Vegetal , Doenças das Plantas/microbiologia , Doenças das Plantas/imunologia , Citrus sinensis/microbiologia , Citrus sinensis/imunologia , Citrus sinensis/metabolismo , Arabidopsis/microbiologia , Arabidopsis/imunologia , Arabidopsis/metabolismo , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/genética , Plantas Geneticamente Modificadas , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Liberibacter/metabolismo , Rhizobiaceae/metabolismo , Nicotiana/microbiologia , Nicotiana/imunologia , Nicotiana/metabolismo , Transporte ProteicoRESUMO
Plants produce reactive oxygen species (ROS) upon infection, which typically trigger defence mechanisms and impede pathogen proliferation. Root-knot nematodes (RKNs, Meloidogyne spp.) represent highly detrimental pathogens capable of parasitizing a broad spectrum of crops, resulting in substantial annual agricultural losses. The involvement of ROS in RKN parasitism is well acknowledged. In this study, we identified a novel effector from Meloidogyne incognita, named CATLe, that contains a conserved catalase domain, exhibiting potential functions in regulating host ROS levels. Phylogenetic analysis revealed that CATLe is conserved across RKNs. Temporal and spatial expression assays showed that the CATLe gene was specifically up-regulated at the early infection stages and accumulated in the subventral oesophageal gland cells of M. incognita. Immunolocalization demonstrated that CATLe was secreted into the giant cells of the host plant during M. incognita parasitism. Transient expression of CATLe significantly dampened the flg22-induced ROS production in Nicotiana benthamiana. In planta assays confirmed that M. incognita can exploit CATLe to manipulate host ROS levels by directly degrading H2O2. Additionally, interfering with expression of the CATLe gene through double-stranded RNA soaking and host-induced gene silencing significantly attenuated M. incognita parasitism, highlighting the important role of CATLe. Taken together, our results suggest that RKNs can directly degrade ROS products using a functional catalase, thereby manipulating host ROS levels and facilitating parasitism.
Assuntos
Catalase , Peróxido de Hidrogênio , Nicotiana , Espécies Reativas de Oxigênio , Tylenchoidea , Animais , Peróxido de Hidrogênio/metabolismo , Tylenchoidea/fisiologia , Espécies Reativas de Oxigênio/metabolismo , Nicotiana/parasitologia , Catalase/metabolismo , Catalase/genética , Doenças das Plantas/parasitologia , Raízes de Plantas/parasitologia , Filogenia , Proteínas de Helminto/metabolismo , Proteínas de Helminto/genética , Interações Hospedeiro-ParasitaRESUMO
Antiviral responses induced by double-stranded RNA (dsRNA) include RNA interference (RNAi) and pattern-triggered immunity (PTI), but their relative contributions to antiviral defence are not well understood. We aimed at testing the impact of exogenous applied dsRNA on both layers of defence against potato virus X expressing GFP (PVX-GFP) in Nicotiana benthamiana. Co-inoculation of PVX-GFP with either sequence-specific (RNAi) or nonspecific dsRNA (PTI) showed that nonspecific dsRNA reduced virus accumulation in both inoculated and systemic leaves. However, nonspecific dsRNA was a poor inducer of antiviral immunity compared to a sequence-specific dsRNA capable of triggering the RNAi response, and plants became susceptible to systemic infection. Studies with a PVX mutant unable to move from cell to cell indicated that the interference with PVX-GFP triggered by nonspecific dsRNA operated at the single-cell level. Next, we performed RNA-seq analysis to examine similarities and differences in the transcriptome triggered by dsRNA alone or in combination with viruses harbouring sequences targeted or not by dsRNA. Enrichment analysis showed an over-representation of plant-pathogen signalling pathways, such as calcium, ethylene and MAPK signalling, which are typical of antimicrobial PTI. Moreover, the transcriptomic response to the virus targeted by dsRNA had a greater impact on defence than the non-targeted virus, highlighting qualitative differences between sequence-specific RNAi and nonspecific PTI responses. Together, these results further our understanding of plant antiviral defence, particularly the contribution of nonspecific dsRNA-mediated PTI. We envisage that both sequence-specific RNAi and nonspecific PTI pathways may be triggered via topical application of dsRNA, contributing cumulatively to plant protection against viruses.
Assuntos
Nicotiana , Doenças das Plantas , Imunidade Vegetal , Potexvirus , Interferência de RNA , RNA de Cadeia Dupla , Nicotiana/virologia , Nicotiana/imunologia , Nicotiana/genética , Imunidade Vegetal/genética , Doenças das Plantas/virologia , Doenças das Plantas/imunologia , Reconhecimento da Imunidade InataRESUMO
Cytidine has a broad range of applications in the pharmaceutical field as an intermediate of antitumor or antiviral agent. Here, a series of new cytidine peptide compounds were synthesized using cytidine and Boc group-protected amino acids and analyzed for their antiviral activities against tobacco mosaic virus (TMV). Among these compounds, the structure of an effective antiviral cytidine peptide SN11 was characterized by 1H NMR, 13C NMR, and high-resolution mass spectrometer. The compound SN11 has a molecular formula of C15H22N6O8 and is named 2-amino-N-(2- ((1- (3,4-dihydroxy-5-(hydroxymethyl) tetrahydrofuran-2-yl) -2-oxo-1,2-dihydropyrimidin-4-yl) amino) -2-oxyethyl) amino). The protection, inactivation, and curation activities of SN11 at a concentration of 500 µg/mL against TMV in Nicotiana glutinosa were 82.6%, 84.2%, and 72.8%, respectively. SN11 also effectively suppressed the systemic transportation of a recombinant TMV carrying GFP reporter gene (p35S-30B:GFP) in Nicotiana benthamiana by reducing viral accumulation to 71.3% in the upper uninoculated leaves and inhibited the systemic infection of TMV in Nicotiana tabacum plants. Furthermore, the results of RNA-seq showed that compound SN11 induced differential expression of genes involved in the biogenesis and function of ribosome, plant hormone signal transduction, plant pathogen interaction, and chromatin. These results validate the antiviral mechanisms of the cytidine peptide compound and provide a theoretical basis for their potential application in the management of plant virus diseases.
Assuntos
Antivirais , Citidina , Nicotiana , Peptídeos , Doenças das Plantas , Vírus do Mosaico do Tabaco , Vírus do Mosaico do Tabaco/efeitos dos fármacos , Antivirais/farmacologia , Antivirais/química , Antivirais/síntese química , Citidina/farmacologia , Citidina/análogos & derivados , Citidina/química , Nicotiana/virologia , Nicotiana/química , Nicotiana/genética , Peptídeos/química , Peptídeos/farmacologia , Peptídeos/síntese química , Doenças das Plantas/virologiaRESUMO
Due to the error-prone nature of viral RNA-dependent RNA polymerases, the replication of RNA viruses results in a diversity of viral genomes harboring point mutations, deletions, insertions, and genome rearrangements. Citrus tristeza virus (CTV), a causal agent of diseases of economically important citrus species, shows intrinsic genetic stability. While the virus appears to have some mechanism that limits the accumulation of single-nucleotide variants, the production of defective viral genomes (DVGs) during virus infection has been reported for certain variants of CTV. The intra-host diversity generated during plant infection with variant T36 (CTV-T36) remains unclear. To address this, we analyzed the RNA species accumulated in the initially infected and systemic leaves of Nicotiana benthamiana plants inoculated with an infectious cDNA clone of CTV-T36, which warranted that infection was initiated by a known, well-defined sequence variant of the virus. CTV-T36 limited the accumulation of single-nucleotide mutants during infection. With that, four types of DVGs-deletions, insertions, and copy- and snap-backs-were found in all the samples, with deletions and insertions being the most common types. Hot-spots across the genome for DVG recombination and short direct sequence repeats suggest that sequence complementarity could mediate DVG formation. In conclusion, our study illustrates the formation of diverse DVGs during CTV-T36 infection. To the best of our knowledge, this is the first study that has analyzed the genetic variability and recombination of a well-defined sequence variant of CTV in an herbaceous host.
Assuntos
Closterovirus , Variação Genética , Genoma Viral , Nicotiana , Doenças das Plantas , RNA Viral , Nicotiana/virologia , Closterovirus/genética , Closterovirus/classificação , Doenças das Plantas/virologia , RNA Viral/genética , Citrus/virologia , Folhas de Planta/virologiaRESUMO
Gene-editing technology, specifically the CRISPR-Cas13a system, has shown promise in breeding plants resistant to RNA viruses. This system targets RNA and, theoretically, can also combat RNA-based viroids. To test this, the CRISPR-Cas13a system was introduced into tomato plants via transient expression and into Nicotiana benthamiana through transgenic methods, using CRISPR RNAs (crRNAs) targeting the conserved regions of both sense and antisense genomes of potato spindle tuber viroid (PSTVd). In tomato plants, the expression of CRISPR-Cas13a and crRNAs substantially reduced PSTVd accumulation and alleviated disease symptoms. In transgenic N. benthamiana plants, the PSTVd levels were lower as compared to wild-type plants. Several effective crRNAs targeting the PSTVd genomic RNA were also identified. These results demonstrate that the CRISPR-Cas13a system can effectively target and combat viroid RNAs, despite their compact structures.
Assuntos
Sistemas CRISPR-Cas , Resistência à Doença , Edição de Genes , Nicotiana , Doenças das Plantas , Plantas Geneticamente Modificadas , Solanum lycopersicum , Viroides , Nicotiana/virologia , Nicotiana/genética , Solanum lycopersicum/virologia , Solanum lycopersicum/genética , Viroides/genética , Doenças das Plantas/virologia , Doenças das Plantas/genética , Edição de Genes/métodos , Plantas Geneticamente Modificadas/virologia , Resistência à Doença/genética , RNA Viral/genética , RNA Viral/metabolismoRESUMO
WRKY transcription factor (TF) plays a crucial role in plant abiotic stress response, but it is rarely reported in Michelia crassipes. Our studies have found that the transcription factor McWRKY43, a member of the IIc subgroup, is strongly upregulated under cold stress. In this study, we cloned the full length of McWRKY43 to further investigate the function of McWRKY43 in resistance to cold stress and its possible regulatory pathways in M. crassipes. Under cold stress, the seed-germination rate of transgenic tobacco was significantly higher than that of the wild type, and the flavonoid content, antioxidant enzyme activities, and proline content of transgenic tobacco seedlings were significantly increased, which promoted the expression of flavonoid pathway structural genes. In addition, the transient transformation of McWRKY43 in the M. crassipes leaves also found the accumulation of flavonoid content and the transcription level of flavonoid structural genes, especially McLDOX, were significantly increased under cold stress. Yeast one-hybrid (Y1H) assay showed that McWRKY43 could bind to McLDOX promoter, and the transcription expression of McLDOX was promoted by McWRKY43 during cold stress treatment. Overall, our results indicated that McWRKY43 is involved in flavonoid biosynthetic pathway to regulate cold stress tolerance of M. crassipes, providing a basis for molecular mechanism of stress resistance in Michelia.
Assuntos
Resposta ao Choque Frio , Flavonoides , Regulação da Expressão Gênica de Plantas , Magnolia , Proteínas de Plantas , Fatores de Transcrição , Temperatura Baixa , Flavonoides/biossíntese , Flavonoides/metabolismo , Nicotiana/genética , Nicotiana/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Magnolia/fisiologiaRESUMO
BACKGROUND: Larch is an important timber tree species. The traditional methods of tree genetic breeding have been progressing slowly. It is necessary to carry out gene function analysis and genetically modified breeding research. The NAC transcription factor family is a plant-specific transcription factor family with various biological functions, as shown in recent research. However, there are few studies on the NAC gene among gymnosperm coniferous species. RESULTS: LoNAC3 with complete cds was identified and isolated from the cDNA of Larix olgensis based on transcriptome data. The cDNA length of LoNAC3 is 1185 bp, encoding 394 amino acids, with a conserved NAM domain located at the N-terminus, and subcellular localization in the nucleus. The results of real-time quantitative PCR analysis showed that at different growth stages and in different tissues of L. olgensis, the relative expression level of LoNAC3 was highest in the needles. After drought, salt, alkali stress and hormone treatment, expression was induced to different degrees. The expression level of LoNAC3 was significantly increased under drought and salt conditions. The relative expression level changed under methyl jasmonate (MeJA) and abscisic acid (ABA) treatment. By observing the phenotype of overexpressed LoNAC3 tobacco, it was found that overexpressed tobacco is shorter and blooms earlier than wild-type tobacco. Under abiotic stress, LoNAC3 overexpressed tobacco has lower germination rates and poorer growth status. Transgenic tobacco under stress treatment has a higher malondialdehyde (MDA) content than wild-type tobacco, while peroxidase (POD) activity is lower than wild-type tobacco. CONCLUSIONS: Through the analysis of LoNAC3 sequence and promoter expression, it can be concluded that LoNAC3 is involved in the drought and salt stress response processes of L. olgensis, and is induced by ABA and MeJA expression. Overexpression of LoNAC3 leads to stunted tobacco growth and negatively regulates its tolerance to drought and salt stress through the reactive oxygen species pathway. The preliminary analysis of the expression pattern and function of the LoNAC3 can provide a theoretical basis and high-quality materials for genetic improvement of larch in later stages.
Assuntos
Larix , Proteínas de Plantas , Fatores de Transcrição , Larix/genética , Larix/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Nicotiana/genética , Regulação da Expressão Gênica de Plantas , Plantas Geneticamente Modificadas/genética , Estresse Fisiológico/genética , Filogenia , Ácido Abscísico/metabolismo , Secas , Genes de Plantas , Acetatos , Ciclopentanos , OxilipinasRESUMO
Oral cancer is the most common malignancy in many developing countries, such as India, due to increased consumption of smokeless tobacco. The trace elemental components in commercially packaged forms of tobacco can play a significant role in the pathogenesis of oral cancer. To qualitatively assess the trace elements in various types of commercially packaged forms of tobacco using laser-induced breakdown spectroscopy (LIBS). Two popular varieties of 'Paan masala' that contained a mixture of slaked lime with areca nut, catechu, and other flavouring agents (tobacco was absent) and four types of packaged tobacco were obtained from 'Paan' shops. The contents in the packets were made into pellets using a hydraulic press and subjected to elemental analysis using LIBS. A ten-trial experiment was carried out on all six pellets. The National Institute of Standards and Technology (NIST) database was used to assess the emission lines. The elements obtained from commercially packaged tobacco and Paan masala were similar: calcium (Ca), iron (Fe), aluminium (Al), nickel (Ni), and chromium (Cr). Substances that cause DNA damage and carcinogenesis are inorganic elements such as nickel. Our study revealed that carcinogens such as nickel are present in the commercially packaged forms of tobacco and 'Paan masala' samples.
Assuntos
Nicotiana , Oligoelementos , Oligoelementos/análise , Nicotiana/química , Análise Espectral/métodos , Níquel/análise , Lasers , Produtos do Tabaco/análise , Embalagem de Produtos , Tabaco sem Fumaça/análise , Cromo/análise , Cálcio/análise , Humanos , Ferro/análiseRESUMO
Wheat leaf blight caused by Bipolaris sorokiniana is a widespread fungal disease that poses a serious risk to wheat. Biological control without causing environmental pollution is one of the safest and most effective method to control plant diseases. The antagonistic bacterial strain HeN-7 (identified as Bacillus velezensis) was isolated from tobacco leaves cultivated in Henan province, China. The results of different concentrations of cell-free supernatant (CFS) from HeN-7 culture against B. sorokiniana mycelia showed that 20% HeN-7 CFS (v/v) reached the maximum inhibition rate of 96%. In the potted plants control assay, B. velezensis HeN-7 CFS exhibited remarkable biocontrol activity on the wheat infected with B. sorokiniana, the best pot control efficacy was 65% at 20% CFS. The research on the mechanism of action demonstrated that HeN-7 CFS induced the membrane lipid peroxidation in B. sorokiniana, leading to the disruption of cell membrane integrity and resulting in the leakage of cell contents; in addition, the intracellular mitochondrial membrane potential in mycelium dissipated and reactive oxygen species accumulated, thereby inhibiting the growth of B. sorokiniana. These results indicate that B. velezensis HeN-7 is a promising candidate as a biological control agent against Bipolaris sorokiniana infection.
Assuntos
Bacillus , Bipolaris , Nicotiana , Doenças das Plantas , Folhas de Planta , Bacillus/isolamento & purificação , Bacillus/metabolismo , Bacillus/fisiologia , Folhas de Planta/microbiologia , Doenças das Plantas/microbiologia , Doenças das Plantas/prevenção & controle , Nicotiana/microbiologia , Triticum/microbiologia , Antifúngicos/farmacologia , Antifúngicos/metabolismo , China , Espécies Reativas de Oxigênio/metabolismo , Micélio/crescimento & desenvolvimento , AntibioseRESUMO
Stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), stands out as one of the most devastating epidemics impacting wheat production worldwide. Resistant wheat varieties had swiftly been overcome due to the emergence of new virulent Pst strains. Effectors secreted by Pst interfere with plant immunity, and verification of their biological function is extremely important for controlling wheat stripe rust. In this study, we identified an effector, Pst-18220, from Puccinia striiformis f. sp. tritici (Pst), which was induced during the early infection stage of Pst. Silencing the expression of Pst-18220 through virus-mediated host-induced gene silencing (HIGS) resulted in a decreased number of rust pustules. In Nicotiana benthamiana, it significantly suppressed cell death induced by Pseudomonas syringae pv. tomato (Pto) DC3000. In Arabidopsis, plants with stable overexpression of Pst-18220 showed increased susceptibility to Pto DC3000, accompanied by a decrease in the expression level of pattern-triggered immunity (PTI)/effector-triggered immunity (ETI)-related genes, namely, AtPCRK1, AtPCRK2, and AtBIK1. These results emphasize the significant role of the Pst candidate effector, Pst-18220, in rust pathogenicity and the suppression of plant defense mechanisms. This broadens our understanding of effectors without any known motif.
Assuntos
Nicotiana , Doenças das Plantas , Puccinia , Triticum , Puccinia/patogenicidade , Doenças das Plantas/microbiologia , Doenças das Plantas/genética , Nicotiana/microbiologia , Nicotiana/genética , Triticum/microbiologia , Pseudomonas syringae/patogenicidade , Arabidopsis/microbiologia , Arabidopsis/genética , Arabidopsis/imunologia , Imunidade Vegetal/genética , Proteínas Fúngicas/metabolismo , Proteínas Fúngicas/genética , Virulência/genética , Resistência à Doença/genéticaRESUMO
Leymus chinensis, a halophytic perennial grass belonging to the Poaceae family, thrives in saline-alkali grasslands and harbors a rich repository of resistance-related genetic resources. This study focused on deciphering the stress-responsive mechanisms of L. chinensis by conducting transcriptomic sequencing under NaHCO3 stress, which resulted in the annotation of a segment corresponding to the 51WRKY gene. The alkali-induced gene LcWRKY40 (QIG37591) was identified by phylogenetic analysis. Real-time quantitative PCR analysis was performed on L. chinensis plants subjected to PEG6000 and alkaline salt (NaHCO3) stress, and the results indicated that the LcWRKY40 gene was upregulated in both the leaves and roots. The localization of the LcWRKY40 protein was confirmed by the use of green fluorescent protein (GFP) fusion technology in transformed rice protoplast cells. The GAL4-driven transformation of the LcWRKY40 gene in INVScI yeast cells, which exhibited enhanced tolerance upon overexpression of the LcWRKY40 gene under mannitol and alkaline salt (NaHCO3) stress conditions. Under drought stress using mannitol, the fresh weight of Nicotiana tabacum overexpressing the LcWRKY40 gene was significantly higher than that of wild-type(WT) tobacco. Through drought and salt alkali stress, we found that overexpressed tobacco at different stages always outperformed the wild type in terms of fresh weight, SOD, MDA, and Fv/Fm. This study provides preliminary insights into the involvement of the LcWRKY40 gene in responding to drought and alkaline salt stresses, highlighting its role in enhancing plant resistance to drought and saline-alkaline conditions. These findings lay the foundation for future molecular breeding strategies aimed at improving grass resistance from different aspects.
Assuntos
Secas , Regulação da Expressão Gênica de Plantas , Nicotiana , Proteínas de Plantas , Tolerância ao Sal , Estresse Fisiológico , Nicotiana/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Estresse Fisiológico/genética , Tolerância ao Sal/genética , Filogenia , Plantas Geneticamente Modificadas/genética , Bicarbonato de Sódio/farmacologia , Poaceae/genética , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Plantas Tolerantes a Sal/genética , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Folhas de Planta/genética , Folhas de Planta/metabolismoRESUMO
After plants transitioned from water to land around 450 million years ago, they faced novel pathogenic microbes. Their colonization of diverse habitats was driven by anatomical innovations like roots, stomata, and vascular tissue, which became central to plant-microbe interactions. However, the impact of these innovations on plant immunity and pathogen infection strategies remains poorly understood. Here, we explore plant-virus interactions in the bryophyte Marchantia polymorpha to gain insights into the evolution of these relationships. Virome analysis reveals that Marchantia is predominantly associated with RNA viruses. Comparative studies with tobacco mosaic virus (TMV) show that Marchantia shares core defense responses with vascular plants but also exhibits unique features, such as a sustained wound response preventing viral spread. Additionally, general defense responses in Marchantia are equivalent to those restricted to vascular tissues in Nicotiana, suggesting that evolutionary acquisition of developmental innovations results in re-routing of defense responses in vascular plants.