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1.
Antonie Van Leeuwenhoek ; 117(1): 92, 2024 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-38949726

RESUMO

Biological control is a promising approach to enhance pathogen and pest control to ensure high productivity in cash crop production. Therefore, PGPR biofertilizers are very suitable for application in the cultivation of tea plants (Camellia sinensis) and tobacco, but it is rarely reported so far. In this study, production of a consortium of three strains of PGPR were applied to tobacco and tea plants. The results demonstrated that plants treated with PGPR exhibited enhanced resistance against the bacterial pathogen Pseudomonas syringae (PstDC3000). The significant effect in improving the plant's ability to resist pathogen invasion was verified through measurements of oxygen activity, bacterial colony counts, and expression levels of resistance-related genes (NPR1, PR1, JAZ1, POD etc.). Moreover, the application of PGPR in the tea plantation showed significantly reduced population occurrences of tea green leafhoppers (Empoasca onukii Matsuda), tea thrips (Thysanoptera:Thripidae), Aleurocanthus spiniferus (Quaintanca) and alleviated anthracnose disease in tea seedlings. Therefore, PGPR biofertilizers may serve as a viable biological control method to improve tobacco and tea plant yield and quality. Our findings revealed part of the mechanism by which PGPR helped improve plant biostresses resistance, enabling better application in agricultural production.


Assuntos
Nicotiana , Controle Biológico de Vetores , Doenças das Plantas , Pseudomonas syringae , Animais , Doenças das Plantas/microbiologia , Doenças das Plantas/prevenção & controle , Nicotiana/microbiologia , Pseudomonas syringae/fisiologia , Controle Biológico de Vetores/métodos , Camellia sinensis/microbiologia , Camellia sinensis/crescimento & desenvolvimento , Insetos/microbiologia , Tisanópteros/microbiologia , Resistência à Doença , Desenvolvimento Vegetal , Agentes de Controle Biológico , Hemípteros/microbiologia
2.
Mol Biol Rep ; 51(1): 791, 2024 Jul 11.
Artigo em Inglês | MEDLINE | ID: mdl-38990430

RESUMO

BACKGROUND: Heat stress is a detrimental abiotic stress that limits the development of many plant species and is linked to a variety of cellular and physiological problems. Heat stress affects membrane fluidity, which leads to negative effects on cell permeability and ion transport. Research reveals that heat stress causes severe damage to cells and leads to rapid accumulation of reactive oxygen species (ROS), which could cause programmed cell death. METHODS AND RESULTS: This current study aimed to validate the role of Triticum aestivum Salt Stress Root Protein (TaSSRP) in plants' tolerance to heat stress by modulating its expression in tobacco plants. The Relative Water Content (RWC), total chlorophyll content, and Membrane Stability Index (MSI) of the seven distinct transgenic lines (T0 - 2, T0 - 3, T0 - 6, T0 - 8, T0 - 9, T0 - 11, and T0 - 13), increased in response to heat stress. Despite the fact that the same tendency was detected in wild-type (WT) plants, changes in physio-biochemical parameters were greater in transgenic lines than in WT plants. The expression analysis revealed that the transgene TaSSRP expressed from 1.00 to 1.809 folds in different lines in the transgenic tobacco plants. The gene TaSSRP offered resistance to heat stress in Nicotiana tabacum, according to the results of the study. CONCLUSION: These findings could help to improve our knowledge and understanding of the mechanism underlying thermotolerance in wheat, and the novel identified gene TaSSRP could be used in generating wheat varieties with enhanced tolerance to heat stress.


Assuntos
Regulação da Expressão Gênica de Plantas , Resposta ao Choque Térmico , Nicotiana , Proteínas de Plantas , Plantas Geneticamente Modificadas , Triticum , Nicotiana/genética , Nicotiana/metabolismo , Triticum/genética , Triticum/metabolismo , Plantas Geneticamente Modificadas/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Resposta ao Choque Térmico/genética , Espécies Reativas de Oxigênio/metabolismo , Raízes de Plantas/metabolismo , Raízes de Plantas/genética , Termotolerância/genética , Clorofila/metabolismo , Tolerância ao Sal/genética
3.
Methods Mol Biol ; 2827: 405-416, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38985285

RESUMO

The engineering of plant cell cultures to produce high-value natural products is suggested to be a safe, low-cost, and environmentally friendly route to produce a wide range of chemicals. Given that the expression of heterologous biosynthetic pathways in plant tissue culture is limited by a lack of detailed protocols, the biosynthesis of high-value metabolites in plant cell culture is constrained compared with that in microbes. However, both Arabidopsis thaliana and Nicotiana benthamiana can be efficiently transformed with multigene constructs to produce high-value natural products in stable plant cell cultures. This chapter provides a detailed protocol as to how to engineer the plant cell culture as bio-factories for metabolite biosynthesis.


Assuntos
Arabidopsis , Produtos Biológicos , Nicotiana , Produtos Biológicos/metabolismo , Nicotiana/metabolismo , Nicotiana/genética , Arabidopsis/metabolismo , Arabidopsis/genética , Técnicas de Cultura de Tecidos/métodos , Células Vegetais/metabolismo , Engenharia Metabólica/métodos , Plantas Geneticamente Modificadas/genética , Metaboloma , Vias Biossintéticas , Metabolômica/métodos , Técnicas de Cultura de Células/métodos
4.
BMC Plant Biol ; 24(1): 655, 2024 Jul 10.
Artigo em Inglês | MEDLINE | ID: mdl-38987695

RESUMO

BACKGROUND: Biochar, a carbon-rich source and natural growth stimulant, is usually produced by the pyrolysis of agricultural biomass. It is widely used to enhance plant growth, enzyme activity, and crop productivity. However, there are no conclusive studies on how different levels of biochar application influence these systems. METHODS AND RESULTS: The present study elucidated the dose-dependent effects of biochar application on the physiological performance, enzyme activity, and dry matter accumulation of tobacco plants via field experiments. In addition, transcriptome analysis was performed on 60-day-old (early growth stage) and 100-day-old (late growth stage) tobacco leaves to determine the changes in transcript levels at the molecular level under various biochar application levels (0, 600, and 1800 kg/ha). The results demonstrated that optimum biochar application enhances plant growth, regulates enzymatic activity, and promotes biomass accumulation in tobacco plants, while higher biochar doses had adverse effects. Furthermore, transcriptome analysis revealed a total of 6561 differentially expressed genes (DEGs) that were up- or down-regulated in the groupwise comparison under different treatments. KEGG pathways analysis demonstrated that carbon fixation in photosynthetic organisms (ko00710), photosynthesis (ko00195), and starch and sucrose metabolism (ko00500) pathways were significantly up-regulated under the optimal biochar dosage (600 kg/ha) and down-regulated under the higher biochar dosage (1800 kg/ha). CONCLUSION: Collectively, these results indicate that biochar application at an optimal rate (600 kg/ha) could positively affect photosynthesis and carbon fixation, which in turn increased the synthesis and accumulation of sucrose and starch, thus promoting the growth and dry matter accumulation of tobacco plants. However, a higher biochar dosage (1800 kg/ha) disturbs the crucial source-sink balance of organic compounds and inhibits the growth of tobacco plants.


Assuntos
Carvão Vegetal , Perfilação da Expressão Gênica , Nicotiana , Nicotiana/genética , Nicotiana/crescimento & desenvolvimento , Nicotiana/efeitos dos fármacos , Transcriptoma , Biomassa , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/genética , Fotossíntese/efeitos dos fármacos
5.
BMC Genomics ; 25(1): 671, 2024 Jul 05.
Artigo em Inglês | MEDLINE | ID: mdl-38970011

RESUMO

BACKGROUND: The dirigent (DIR) genes encode proteins that act as crucial regulators of plant lignin biosynthesis. In Solanaceae species, members of the DIR gene family are intricately related to plant growth and development, playing a key role in responding to various biotic and abiotic stresses. It will be of great application significance to analyze the DIR gene family and expression profile under various pathogen stresses in Solanaceae species. RESULTS: A total of 57 tobacco NtDIRs and 33 potato StDIRs were identified based on their respective genome sequences. Phylogenetic analysis of DIR genes in tobacco, potato, eggplant and Arabidopsis thaliana revealed three distinct subgroups (DIR-a, DIR-b/d and DIR-e). Gene structure and conserved motif analysis showed that a high degree of conservation in both exon/intron organization and protein motifs among tobacco and potato DIR genes, especially within members of the same subfamily. Total 8 pairs of tandem duplication genes (3 pairs in tobacco, 5 pairs in potato) and 13 pairs of segmental duplication genes (6 pairs in tobacco, 7 pairs in potato) were identified based on the analysis of gene duplication events. Cis-regulatory elements of the DIR promoters participated in hormone response, stress responses, circadian control, endosperm expression, and meristem expression. Transcriptomic data analysis under biotic stress revealed diverse response patterns among DIR gene family members to pathogens, indicating their functional divergence. After 96 h post-inoculation with Ralstonia solanacearum L. (Ras), tobacco seedlings exhibited typical symptoms of tobacco bacterial wilt. The qRT-PCR analysis of 11 selected NtDIR genes displayed differential expression pattern in response to the bacterial pathogen Ras infection. Using line 392278 of potato as material, typical symptoms of potato late blight manifested on the seedling leaves under Phytophthora infestans infection. The qRT-PCR analysis of 5 selected StDIR genes showed up-regulation in response to pathogen infection. Notably, three clustered genes (NtDIR2, NtDIR4, StDIR3) exhibited a robust response to pathogen infection, highlighting their essential roles in disease resistance. CONCLUSION: The genome-wide identification, evolutionary analysis, and expression profiling of DIR genes in response to various pathogen infection in tobacco and potato have provided valuable insights into the roles of these genes under various stress conditions. Our results could provide a basis for further functional analysis of the DIR gene family under pathogen infection conditions.


Assuntos
Evolução Molecular , Família Multigênica , Nicotiana , Filogenia , Proteínas de Plantas , Solanum tuberosum , Solanum tuberosum/genética , Solanum tuberosum/microbiologia , Nicotiana/genética , Nicotiana/microbiologia , Proteínas de Plantas/genética , Regulação da Expressão Gênica de Plantas , Doenças das Plantas/microbiologia , Doenças das Plantas/genética , Estresse Fisiológico/genética , Regiões Promotoras Genéticas , Duplicação Gênica , Ralstonia solanacearum , Genes de Plantas
6.
Sci Rep ; 14(1): 15309, 2024 07 03.
Artigo em Inglês | MEDLINE | ID: mdl-38961197

RESUMO

Axillary bud is an important aspect of plant morphology, contributing to the final tobacco yield. However, the mechanisms of axillary bud development in tobacco remain largely unknown. To investigate this aspect of tobacco biology, the metabolome and proteome of the axillary buds before and after topping were compared. A total of 569 metabolites were differentially abundant before and 1, 3, and 5 days after topping. KEGG analyses further revealed that the axillary bud was characterized by a striking enrichment of metabolites involved in flavonoid metabolism, suggesting a strong flavonoid biosynthesis activity in the tobacco axillary bud after topping. Additionally, 9035 differentially expressed proteins (DEPs) were identified before and 1, 3, and 5 days after topping. Subsequent GO and KEGG analyses revealed that the DEPs in the axillary bud were enriched in oxidative stress, hormone signal transduction, MAPK signaling pathway, and starch and sucrose metabolism. The integrated proteome and metabolome analysis revealed that the indole-3-acetic acid (IAA) alteration in buds control dormancy release and sustained growth of axillary bud by regulating proteins involved in carbohydrate metabolism, amino acid metabolism, and lipid metabolism. Notably, the proteins related to reactive oxygen species (ROS) scavenging and flavonoid biosynthesis were strongly negatively correlated with IAA content. These findings shed light on a critical role of IAA alteration in regulating axillary bud outgrowth, and implied a potential crosstalk among IAA alteration, ROS homeostasis, and flavonoid biosynthesis in tobacco axillary bud under topping stress, which could improve our understanding of the IAA alteration in axillary bud as an important regulator of axillary bud development.


Assuntos
Ácidos Indolacéticos , Metaboloma , Nicotiana , Proteínas de Plantas , Proteoma , Ácidos Indolacéticos/metabolismo , Nicotiana/metabolismo , Nicotiana/crescimento & desenvolvimento , Proteoma/metabolismo , Proteínas de Plantas/metabolismo , Regulação da Expressão Gênica de Plantas , Flavonoides/metabolismo , Flores/metabolismo , Flores/crescimento & desenvolvimento , Reguladores de Crescimento de Plantas/metabolismo
7.
Sci Adv ; 10(28): eadl3591, 2024 Jul 12.
Artigo em Inglês | MEDLINE | ID: mdl-38985863

RESUMO

The hydrogen isotopic composition (δ2H) of plant compounds is increasingly used as a hydroclimatic proxy; however, the interpretation of δ2H values is hampered by potential coeffecting biochemical and biophysical processes. Here, we studied δ2H values of water and carbohydrates in leaves and roots, and of leaf n-alkanes, in two distinct tobacco (Nicotiana sylvestris) experiments. Large differences in plant performance and biochemistry resulted from (a) soil fertilization with varying nitrogen (N) species ratios and (b) knockout-induced starch deficiency. We observed a strong 2H-enrichment in sugars and starch with a decreasing performance induced by increasing NO3-/NH4+ ratios and starch deficiency, as well as from leaves to roots. However, δ2H values of cellulose and n-alkanes were less affected. We show that relative concentrations of sugars and starch, interlinked with leaf gas exchange, shape δ2H values of carbohydrates. We thus provide insights into drivers of hydrogen isotopic composition of plant compounds and into the mechanistic modeling of plant cellulose δ2H values.


Assuntos
Carboidratos , Hidrogênio , Folhas de Planta , Folhas de Planta/química , Folhas de Planta/metabolismo , Hidrogênio/análise , Carboidratos/química , Carboidratos/análise , Amido/química , Nicotiana/química , Lipídeos/análise , Lipídeos/química , Raízes de Plantas/química , Raízes de Plantas/metabolismo , Metabolismo dos Carboidratos , Deutério/química , Alcanos/análise , Alcanos/química , Água/química
9.
Planta ; 260(2): 41, 2024 Jul 02.
Artigo em Inglês | MEDLINE | ID: mdl-38954109

RESUMO

MAIN CONCLUSION: In this study, six ZaBZRs were identified in Zanthoxylum armatum DC, and all the ZaBZRs were upregulated by abscisic acid (ABA) and drought. Overexpression of ZaBZR1 enhanced the drought tolerance of transgenic Nicotiana benthamian. Brassinosteroids (BRs) are a pivotal class of sterol hormones in plants that play a crucial role in plant growth and development. BZR (brassinazole resistant) is a crucial transcription factor in the signal transduction pathway of BRs. However, the BZR gene family members have not yet been identified in Zanthoxylum armatum DC. In this study, six members of the ZaBZR family were identified by bioinformatic methods. All six ZaBZRs exhibited multiple phosphorylation sites. Phylogenetic and collinearity analyses revealed a closest relationship between ZaBZRs and ZbBZRs located on the B subgenomes. Expression analysis revealed tissue-specific expression patterns of ZaBZRs in Z. armatum, and their promoter regions contained cis-acting elements associated with hormone response and stress induction. Additionally, all six ZaBZRs showed upregulation upon treatment after abscisic acid (ABA) and polyethylene glycol (PEG), indicating their participation in drought response. Subsequently, we conducted an extensive investigation of ZaBZR1. ZaBZR1 showed the highest expression in the root, followed by the stem and terminal bud. Subcellular localization analysis revealed that ZaBZR1 is present in the cytoplasm and nucleus. Overexpression of ZaBZR1 in transgenic Nicotiana benthamiana improved seed germination rate and root growth under drought conditions, reducing water loss rates compared to wild-type plants. Furthermore, ZaBZR1 increased proline content (PRO) and decreased malondialdehyde content (MDA), indicating improved tolerance to drought-induced oxidative stress. The transgenic plants also showed a reduced accumulation of reactive oxygen species. Importantly, ZaBZR1 up-regulated the expression of drought-related genes such as NbP5CS1, NbDREB2A, and NbWRKY44. These findings highlight the potential of ZaBZR1 as a candidate gene for enhancing drought resistance in transgenic N. benthamiana and provide insight into the function of ZaBZRs in Z. armatum.


Assuntos
Secas , Regulação da Expressão Gênica de Plantas , Filogenia , Proteínas de Plantas , Plantas Geneticamente Modificadas , Zanthoxylum , Plantas Geneticamente Modificadas/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Zanthoxylum/genética , Zanthoxylum/fisiologia , Zanthoxylum/metabolismo , Nicotiana/genética , Nicotiana/fisiologia , Nicotiana/efeitos dos fármacos , Ácido Abscísico/metabolismo , Ácido Abscísico/farmacologia , Família Multigênica , Brassinosteroides/metabolismo , Brassinosteroides/farmacologia , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Estresse Fisiológico/genética , Reguladores de Crescimento de Plantas/metabolismo , Reguladores de Crescimento de Plantas/farmacologia , Resistência à Seca
10.
Mol Plant Pathol ; 25(7): e13491, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38961768

RESUMO

Root-knot nematodes (RKNs) are microscopic parasitic worms able to infest the roots of thousands of plant species, causing massive crop yield losses worldwide. They evade the plant's immune system and manipulate plant cell physiology and metabolism to transform a few root cells into giant cells, which serve as feeding sites for the nematode. RKN parasitism is facilitated by the secretion in planta of effector molecules, mostly proteins that hijack host cellular processes. We describe here a conserved RKN-specific effector, effector 12 (EFF12), that is synthesized exclusively in the oesophageal glands of the nematode, and we demonstrate its function in parasitism. In the plant, MiEFF12 localizes to the endoplasmic reticulum (ER). A combination of RNA-sequencing analysis and immunity-suppression bioassays revealed the contribution of MiEFF12 to the modulation of host immunity. Yeast two-hybrid, split luciferase and co-immunoprecipitation approaches identified an essential component of the ER quality control system, the Solanum lycopersicum plant bap-like (PBL), and basic leucine zipper 60 (BZIP60) proteins as host targets of MiEFF12. Finally, silencing the PBL genes in Nicotiana benthamiana decreased susceptibility to Meloidogyne incognita infection. Our results suggest that EFF12 manipulates PBL function to modify plant immune responses to allow parasitism.


Assuntos
Retículo Endoplasmático , Tylenchoidea , Animais , Retículo Endoplasmático/metabolismo , Tylenchoidea/fisiologia , Tylenchoidea/patogenicidade , Proteínas de Helminto/metabolismo , Proteínas de Helminto/genética , Imunidade Vegetal , Nicotiana/parasitologia , Nicotiana/imunologia , Nicotiana/genética , Solanum lycopersicum/parasitologia , Solanum lycopersicum/imunologia , Solanum lycopersicum/genética , Doenças das Plantas/parasitologia , Doenças das Plantas/imunologia , Raízes de Plantas/parasitologia , Raízes de Plantas/imunologia , Interações Hospedeiro-Parasita
11.
Multimedia | Recursos Multimídia | ID: multimedia-13199

RESUMO

Dejar de fumar y vapear puede ser un proceso difícil, y a veces no es posible conseguirlo sin ayuda.


Assuntos
Nicotiana , Vaping
12.
Plant Cell Rep ; 43(7): 177, 2024 Jun 19.
Artigo em Inglês | MEDLINE | ID: mdl-38898307

RESUMO

KEY MESSAGE: Recently published high-quality reference genome assemblies indicate that, in addition to RDR1-deficiency, the loss of several key RNA silencing-associated genes may contribute to the hypersusceptibility of Nicotiana benthamiana to viruses.


Assuntos
Nicotiana , Doenças das Plantas , Interferência de RNA , Nicotiana/genética , Nicotiana/virologia , Doenças das Plantas/virologia , Doenças das Plantas/genética , Vírus de Plantas/fisiologia , Vírus de Plantas/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Genes de Plantas/genética , Regulação da Expressão Gênica de Plantas
13.
BMC Plant Biol ; 24(1): 583, 2024 Jun 20.
Artigo em Inglês | MEDLINE | ID: mdl-38898384

RESUMO

BACKGROUND: Leaf morphology plays a crucial role in photosynthetic efficiency and yield potential in crops. Cigar tobacco plants, which are derived from common tobacco (Nicotiana tabacum L.), possess special leaf characteristics including thin and delicate leaves with few visible veins, making it a good system for studying the genetic basis of leaf morphological characters. RESULTS: In this study, GWAS and QTL mapping were simultaneously performed using a natural population containing 185 accessions collected worldwide and an F2 population consisting of 240 individuals, respectively. A total of 26 QTLs related to leaf morphological traits were mapped in the F2 population at three different developmental stages, and some QTL intervals were repeatedly detected for different traits and at different developmental stages. Among the 206 significant SNPs identified in the natural population using GWAS, several associated with the leaf thickness phenotype were co-mapped via QTL mapping. By analyzing linkage disequilibrium and transcriptome data from different tissues combined with gene functional annotations, 7 candidate genes from the co-mapped region were identified as the potential causative genes associated with leaf thickness. CONCLUSIONS: These results presented a valuable cigar tobacco resource showing the genetic diversity regarding its leaf morphological traits at different developmental stages. It also provides valuable information for novel genes and molecular markers that will be useful for further functional verification and for molecular breeding of leaf morphological traits in crops in the future.


Assuntos
Mapeamento Cromossômico , Estudo de Associação Genômica Ampla , Nicotiana , Folhas de Planta , Locos de Características Quantitativas , Nicotiana/genética , Nicotiana/anatomia & histologia , Nicotiana/crescimento & desenvolvimento , Folhas de Planta/anatomia & histologia , Folhas de Planta/genética , Folhas de Planta/crescimento & desenvolvimento , Fenótipo , Polimorfismo de Nucleotídeo Único , Desequilíbrio de Ligação
14.
Int J Mol Sci ; 25(11)2024 May 31.
Artigo em Inglês | MEDLINE | ID: mdl-38892274

RESUMO

Heavy metals are dangerous contaminants that constitute a threat to human health because they persist in soils and are easily transferred into the food chain, causing damage to human health. Among heavy metals, nickel appears to be one of the most dangerous, being responsible for different disorders. Public health protection requires nickel detection in the environment and food chains. Biosensors represent simple, rapid, and sensitive methods for detecting nickel contamination. In this paper, we report on the setting up a whole-cell-based system, in which protoplasts, obtained from Nicotiana tabacum leaves, were used as transducers to detect the presence of heavy metal ions and, in particular, nickel ions. Protoplasts were genetically modified with a plasmid containing the Green Fluorescent Protein reporter gene (GFP) under control of the promoter region of a sunflower gene coding for a small Heat Shock Protein (HSP). Using this device, the presence of heavy metal ions was detected. Thus, the possibility of using this whole-cell system as a novel tool to detect the presence of nickel ions in food matrices was assessed.


Assuntos
Técnicas Biossensoriais , Níquel , Nicotiana , Protoplastos , Níquel/análise , Protoplastos/metabolismo , Nicotiana/genética , Técnicas Biossensoriais/métodos , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Contaminação de Alimentos/análise , Metais Pesados/análise
15.
Plant Signal Behav ; 19(1): 2359257, 2024 Dec 31.
Artigo em Inglês | MEDLINE | ID: mdl-38825861

RESUMO

Potassium (K+) plays a role in enzyme activation, membrane transport, and osmotic regulation processes. An increase in potassium content can significantly improve the elasticity and combustibility of tobacco and reduce the content of harmful substances. Here, we report that the expression analysis of Nt GF14e, a 14-3-3 gene, increased markedly after low-potassium treatment (LK). Then, chlorophyll content, POD activity and potassium content, were significantly increased in overexpression of Nt GF14e transgenic tobacco lines compared with those in the wild type plants. The net K+ efflux rates were severely lower in the transgenic plants than in the wild type under LK stress. Furthermore, transcriptome analysis identified 5708 upregulated genes and 2787 downregulated genes between Nt GF14e overexpressing transgenic tobacco plants. The expression levels of some potassium-related genes were increased, such as CBL-interacting protein kinase 2 (CIPK2), Nt CIPK23, Nt CIPK25, H+-ATPase isoform 2 a (AHA2a), Nt AHA4a, Stelar K+ outward rectifier 1(SKOR1), and high affinity K+ transporter 5 (HAK5). The result of yeast two-hybrid and luciferase complementation imaging experiments suggested Nt GF14e could interact with CIPK2. Overall, these findings indicate that NtGF14e plays a vital roles in improving tobacco LK tolerance and enhancing potassium nutrition signaling pathways in tobacco plants.


Assuntos
Proteínas 14-3-3 , Regulação da Expressão Gênica de Plantas , Nicotiana , Proteínas de Plantas , Plantas Geneticamente Modificadas , Potássio , Nicotiana/genética , Nicotiana/metabolismo , Proteínas 14-3-3/metabolismo , Proteínas 14-3-3/genética , Potássio/metabolismo , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas/metabolismo , Estresse Fisiológico/genética
16.
J Physiol Sci ; 74(1): 35, 2024 Jun 25.
Artigo em Inglês | MEDLINE | ID: mdl-38918702

RESUMO

BACKGROUND: The increasing prevalence of heated tobacco products (HTPs) has heightened concerns regarding their potential health risks. Previous studies have demonstrated the toxicity of cigarette smoke extract (CSE) from traditional tobacco's mainstream smoke, even after the removal of nicotine and tar. Our study aimed to investigate the cytotoxicity of CSE derived from HTPs and traditional tobacco, with a particular focus on the role of reactive oxygen species (ROS) and intracellular Ca2+. METHODS: A human oral squamous cell carcinoma (OSCC) cell line, HSC-3 was utilized. To prepare CSE, aerosols from HTPs (IQOS) and traditional tobacco products (1R6F reference cigarette) were collected into cell culture media. A cell viability assay, apoptosis assay, western blotting, and Fluo-4 assay were conducted. Changes in ROS levels were measured using electron spin resonance spectroscopy and the high-sensitivity 2',7'-dichlorofluorescein diacetate assay. We performed a knockdown of calcium/calmodulin-dependent protein kinase kinase 2 (CaMKK2) by shRNA lentivirus in OSCC cells. RESULTS: CSE from both HTPs and traditional tobacco exhibited cytotoxic effects in OSCC cells. Exposure to CSE from both sources led to an increase in intracellular Ca2+ concentration and induced p38 phosphorylation. Additionally, these extracts prompted cell apoptosis and heightened ROS levels. N-acetylcysteine (NAC) mitigated the cytotoxic effects and p38 phosphorylation. Furthermore, the knockdown of CaMKK2 in HSC-3 cells reduced cytotoxicity, ROS production, and p38 phosphorylation in response to CSE. CONCLUSION: Our findings suggest that the CSE from both HTPs and traditional tobacco induce cytotoxicity. This toxicity is mediated by ROS, which are regulated through Ca2+ signaling and CaMKK2 pathways.


Assuntos
Quinase da Proteína Quinase Dependente de Cálcio-Calmodulina , Carcinoma de Células Escamosas , Neoplasias Bucais , Espécies Reativas de Oxigênio , Fumaça , Produtos do Tabaco , Humanos , Espécies Reativas de Oxigênio/metabolismo , Neoplasias Bucais/metabolismo , Neoplasias Bucais/patologia , Linhagem Celular Tumoral , Fumaça/efeitos adversos , Carcinoma de Células Escamosas/metabolismo , Quinase da Proteína Quinase Dependente de Cálcio-Calmodulina/metabolismo , Produtos do Tabaco/efeitos adversos , Apoptose/efeitos dos fármacos , Nicotiana/química , Cálcio/metabolismo , Sobrevivência Celular/efeitos dos fármacos
17.
Biomolecules ; 14(6)2024 Jun 16.
Artigo em Inglês | MEDLINE | ID: mdl-38927115

RESUMO

Resveratrol, a phenylpropanoid compound, exhibits diverse pharmacological properties, making it a valuable candidate for health and disease management. However, the demand for resveratrol exceeds the capacity of plant extraction methods, necessitating alternative production strategies. Microbial synthesis offers several advantages over plant-based approaches and presents a promising alternative. Yarrowia lipolytica stands out among microbial hosts due to its safe nature, abundant acetyl-CoA and malonyl-CoA availability, and robust pentose phosphate pathway. This study aimed to engineer Y. lipolytica for resveratrol production. The resveratrol biosynthetic pathway was integrated into Y. lipolytica by adding genes encoding tyrosine ammonia lyase from Rhodotorula glutinis, 4-coumarate CoA ligase from Nicotiana tabacum, and stilbene synthase from Vitis vinifera. This resulted in the production of 14.3 mg/L resveratrol. A combination of endogenous and exogenous malonyl-CoA biosynthetic modules was introduced to enhance malonyl-CoA availability. This included genes encoding acetyl-CoA carboxylase 2 from Arabidopsis thaliana, malonyl-CoA synthase, and a malonate transporter protein from Bradyrhizobium diazoefficiens. These strategies increased resveratrol production to 51.8 mg/L. The further optimization of fermentation conditions and the utilization of sucrose as an effective carbon source in YP media enhanced the resveratrol concentration to 141 mg/L in flask fermentation. By combining these strategies, we achieved a titer of 400 mg/L resveratrol in a controlled fed-batch bioreactor. These findings demonstrate the efficacy of Y. lipolytica as a platform for the de novo production of resveratrol and highlight the importance of metabolic engineering, enhancing malonyl-CoA availability, and media optimization for improved resveratrol production.


Assuntos
Engenharia Metabólica , Resveratrol , Sacarose , Yarrowia , Resveratrol/metabolismo , Yarrowia/genética , Yarrowia/metabolismo , Engenharia Metabólica/métodos , Sacarose/metabolismo , Aciltransferases/genética , Aciltransferases/metabolismo , Vitis/microbiologia , Vitis/genética , Vitis/metabolismo , Coenzima A Ligases/metabolismo , Coenzima A Ligases/genética , Malonil Coenzima A/metabolismo , Nicotiana/genética , Nicotiana/metabolismo , Nicotiana/microbiologia , Rhodotorula/genética , Rhodotorula/metabolismo , Fermentação , Arabidopsis/genética , Arabidopsis/metabolismo , Amônia-Liases , Proteínas de Bactérias
18.
Plant Mol Biol ; 114(4): 73, 2024 Jun 14.
Artigo em Inglês | MEDLINE | ID: mdl-38874648

RESUMO

Functional genomics through transgenesis has provided faster and more reliable methods for identifying, characterizing, and utilizing genes or quantitative trait loci linked to agronomic traits to target yield. The present study explored the role of Big Grain1 (BG1) gene of rice (Oryza sativa L.) in yield improvement of crop plants. We aimed to identify the genetic variation of OsBG1 in various indica rice cultivars by studying the allelic polymorphism of the gene, while also investigating the gene's potential to increase crop yield through the transgenic approach. Our study reports the presence of an extra 393 bp sequence having two 6 bp enhancer elements in the 3' regulatory sequence of OsBG1 in the large-grain cultivar IR64 but not in the small-grain cultivar Badshahbhog. A single copy of the OsBG1 gene in both the cultivars and a 4.1-fold higher expression of OsBG1 in IR64 than in Badshahbhog imply that the grain size is positively correlated with the level of OsBG1 expression in rice. The ectopic expression of OsBG1 under the endosperm-specific glutelin C promoter in Badshahbhog enhanced the flag leaf length, panicle weight, and panicle length by an average of 33.2%, 33.7%, and 30.5%, respectively. The length of anthers, spikelet fertility, and grain yield per plant increased in transgenic rice lines by an average of 27.5%, 8.3%, and 54.4%, respectively. Heterologous expression of OsBG1 under the constitutive 2xCaMV35S promoter improved the number of seed pods per plant and seed yield per plant in transgenic tobacco lines by an average of 2.2-fold and 2.6-fold, respectively. Improving crop yield is crucial to ensure food security and socio-economic stability, and identifying suitable genetic factor is the essential step towards this endeavor. Our findings suggest that the OsBG1 gene is a promising candidate for improving the grain yield of monocot and dicot plant systems by molecular breeding and genetic engineering.


Assuntos
Grão Comestível , Regulação da Expressão Gênica de Plantas , Nicotiana , Oryza , Proteínas de Plantas , Plantas Geneticamente Modificadas , Oryza/genética , Oryza/crescimento & desenvolvimento , Nicotiana/genética , Nicotiana/crescimento & desenvolvimento , Grão Comestível/genética , Grão Comestível/crescimento & desenvolvimento , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Regiões Promotoras Genéticas/genética , Sementes/genética , Sementes/crescimento & desenvolvimento
19.
Mol Plant Pathol ; 25(6): e13487, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38877765

RESUMO

We had previously reported that a plum pox virus (PPV)-based chimera that had its P1-HCPro bi-cistron replaced by a modified one from potato virus Y (PVY) increased its virulence in some Nicotiana benthamiana plants, after mechanical passages. This correlated with the natural acquisition of amino acid substitutions in several proteins, including in HCPro at either position 352 (Ile→Thr) or 454 (Leu→Arg), or of mutations in non-coding regions. Thr in position 352 is not found among natural potyviruses, while Arg in 454 is a reversion to the native PVY HCPro amino acid. We show here that both mutations separately contributed to the increased virulence observed in the passaged chimeras that acquired them, and that Thr in position 352 is no intragenic suppressor to a Leu in position 454, because their combined effects were cumulative. We demonstrate that Arg in position 454 improved HCPro autocatalytic cleavage, while Thr in position 352 increased its accumulation and the silencing suppression of a reporter in agropatch assays. We assessed infection by four cloned chimera variants expressing HCPro with none of the two substitutions, one of them or both, in wild-type versus DCL2/4-silenced transgenic plants. We found that during infection, the transgenic context of altered small RNAs affected the accumulation of the four HCPro variants differently and hence, also infection virulence.


Assuntos
Substituição de Aminoácidos , Nicotiana , Potyvirus , Proteínas Virais , Virulência/genética , Nicotiana/virologia , Potyvirus/patogenicidade , Potyvirus/genética , Proteínas Virais/metabolismo , Proteínas Virais/genética , Doenças das Plantas/virologia , Quimera , Vírus Eruptivo da Ameixa/patogenicidade , Vírus Eruptivo da Ameixa/genética , Cisteína Endopeptidases/metabolismo , Cisteína Endopeptidases/genética , Mutação/genética
20.
Pestic Biochem Physiol ; 202: 105896, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38879345

RESUMO

The objective of this study was to investigate the mechanism underlying LW-1-induced resistance to TMV in wild-type and salicylic acid (SA)-deficient NahG transgenic tobacco plants. Our findings revealed that LW-1 failed to induce antivirus infection activity and increase SA content in NahG tobacco, indicating the crucial role of SA in these processes. Meanwhile, LW-1 triggered defense-related early-signaling nitric oxide (NO) generation, as evidenced by the emergence of NO fluorescence in both types of tobacco upon treatment with LW-1, however, NO fluorescence was stronger in NahG compared to wild-type tobacco. Notably, both of them were eliminated by the NO scavenger cPTIO, which also reversed LW-1-induced antivirus activity and the increase of SA content, suggesting that NO participates in LW-1-induced resistance to TMV, and may act upstream of the SA pathway. Defense-related enzymes and genes were detected in tobacco with or without TMV inoculation, and the results showed that LW-1 regulated both enzyme activity (ß-1,3-glucanase [GLU], catalase [CAT] and phenylalanine ammonia-lyase [PAL]) and gene expression (PR1, PAL, WYKY4) through NO signaling in both SA-dependent and SA-independent pathways.


Assuntos
Resistência à Doença , Nicotiana , Óxido Nítrico , Doenças das Plantas , Ácido Salicílico , Vírus do Mosaico do Tabaco , Nicotiana/metabolismo , Nicotiana/genética , Ácido Salicílico/metabolismo , Ácido Salicílico/farmacologia , Óxido Nítrico/metabolismo , Plantas Geneticamente Modificadas , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Transdução de Sinais , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos
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