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Les systèmes de guérison traditionnels du Pérou – dont certains remonteraient à l’époque préhistorique, selon les estimations – sont reconnus pour leur utilisation saillante des plantes psychotropes. En outre, les traditions de soins rattachées aux montagnes andines, appelées sierra, s’articulent autour du cocaïer (Erythroxylum coca). Pour leur part, les traditions de guérison qui proviennent des déserts de la côte pacifique du nord (costa) sont plutôt caractérisées par l’utilisation du cactus psychoactif huachuma (Echinopsis pachanoi). Finalement les systèmes médicaux qui se sont développés à l’est des Andes, dans les forêts tropicales de l’Amazonie, font appel à diverses plantes psychotropes, dont le tabac (Nicotiana rustica) et l’ayahuasca (Banisteriopsis caapi) sont les plus connus. Le présent article porte sur les traditions de guérison péruviennes, en particulier la branche amazonienne, dans le contexte de la renaissance psychédélique actuelle. Nous soutenons que l’inclusivité culturelle des études cliniques dans le cadre de la reprise des recherches psychédéliques est indispensable, en présentant deux exemples de recherche transculturelle clinique sur le terrain. Le premier inclut une étude collaborative effectuée avec un guérisseur traditionnel amazonien qui se spécialise dans l’utilisation de la plante de tabac à des fins thérapeutiques, tandis que le second met l’accent sur un programme intégratif de traitement des toxicomanies combinant médecine amazonienne et psychothérapie. Ces exemples illustrent des moyens thérapeutiques prometteurs soulignant l’utilité de l’approche transculturelle, non seulement dans le cadre clinique, mais aussi pour l’équité culturelle dans la renaissance psychédélique., Peruvian traditional healing systems, some of which have been estimated to date back to prehistoric times, are known for their prominent usage of psychoactive plants: For curing traditions in the Andean highlands or sierra, it is the coca bush (Erythroxylum coca) that plays the protagonist role. Healing traditions that have arisen in the deserts of the northern Pacific coast (costa), on the other hand, are characterized by the usage of the psychoactive huachuma cactus (Echinopsis pachanoi). Finally, medical systems that have evolved east to the Andes in the tropical forests of the Amazon make use of diverse psychoactive plants, of which especially tobacco (Nicotiana rustica) and ayahuasca (Banisteriopsis caapi) are known. The present contribution takes reference to the Peruvian healing tradition and mainly its Amazonian branch in the context of the ongoing psychedelic renaissance. We argue that culturally inclusive studies are critical in the scope of the revival of psychedelic research and present two examples of cross-cultural clinical field research in this context. The first one involved a collaborative study with an Amazonian traditional healer specializing in the usage of the tobacco plant for therapeutic purposes, while the second one focused on an integrative Peruvian addiction treatment program, in which Amazonian medicine is combined with psychotherapy. The examples point to promising therapeutic means and underscore the importance of an intercultural approach in view of both clinical utility and cultural equality in the psychedelic renaissance., Los sistemas de curación peruanos, algunos de los cuales se estima que existen desde tiempos prehistóricos, son conocidos por su prominente uso de plantas psicoactivas: Para las tradiciones de la sierra andina, es la planta de la coca (Erythroxylum coca) la que juega el rol protagonista. Por otra parte, las tradiciones curativas que surgieron en los desiertos de la costa pacífica del norte, se caracterizan por el uso de un cactus psicoactivo llamado huachuma (Echinopsis pachanoi). Finalmente, los sistemas de curación que evolucionaron al este de los Andes, en los bosques tropicales de la Amazonía, hacen uso de diversas plantas psicoactivas, de los cuales son especialmente conocidos el tabaco (Nicotiana rustica) y la ayahuasca (Banisteriopsis caapi). El presente artículo hace referencia a la tradición curativa peruana, y principalmente a su rama amazónica, en el contexto del renacimiento psicodélico. Argumentamos que en el ámbito actual del resurgimiento de la investigación científica sobre sustancias psicoactivas, los estudios culturalmente incluyentes son fundamentales. En ese contexto presentamos dos ejemplos de estudios de campo clínicos interculturales. El primero involucra un estudio transdiciplinario colaborativo que incluye un curandero amazónico especializado en el uso de la planta del tabaco para fines terapéuticos. El secundo se enfoca en un programa de tratamiento de adiciones integrativo en la Amazonía peruana, el cual combina la medicina amazónica con psicoterapia. Los ejemplos apuntan a sistemas terapéuticos prometedores y subrayan la importancia de un abordaje intercultural, tanto a la vista de la utilidad clínica, como de la equidad cultural en el renacimiento psicodélico.

Host resistance is an important tool in the management of black shank disease of tobacco. Race development leads to rapid loss of single-gene resistance, but the adaptation by Phytophthora nicotianae to sources of partial resistance from Beinhart 1000, Florida 301, and the Wz gene region introgressed from Nicotiana rustica is poorly characterized. In greenhouse environments, host genotypes with quantitative trait loci (QTLs) conferring resistance from multiple sources were initially inoculated with an aggressive isolate of race 0 or race 1 of P. nicotianae. The most aggressive isolate was selected after each of six host generations to inoculate the next generation of plants. The race 0 isolate demonstrated a continuous gradual increase in disease severity and percentage root rot on all sources of resistance except the genotype K 326 Wz/–, where a large increase in both was observed between generations 2 and 3. Adaptation by the race 0 isolate on Beinhart 1000 represents the first report of adaptation to this genotype by P. nicotianae. The race 1 isolate did not exhibit significant increases in aggressiveness over generations but exhibited a large increase in aggressiveness on K 326 Wz/– between generations 3 and 4. Molecular characterization of isolates recovered during selection was completed via double digest restriction-site associated DNA sequencing, but no polymorphisms were associated with the observed changes in aggressiveness. The rapid adaptation to Wz resistance and the gradual adaptation to other QTLs highlights the need to study the nature of Wz resistance and to conduct field studies on the efficacy of resistance gene rotation for disease management.

Nitrik oksit, hidrokarbonlar, aldehidler, fenollar, kinon ve semikinon radikalleri gibi pek çok kimyasal maddeyi bünyesinde barındıran ve doğrudan ya da dolaylı olarak oksijen kaynaklı serbest radikal oluşumuna yol açan tütün keşfedilmesinden günümüze kadar geçen zaman içinde yaygın olarak kullanılmaktadır. Bu yüzden dolayı Nicotiana rustica L.’nin fenolik içeriğinin, antioksidan/oksidan özelliklerinin bilinmesi ve önemli metabolik enzim olan Paraoksonaz (PON 1) ve Asetilkolinesteraz (AChE) üzerine etki çalışmaları önem arzetmektedir. Çalışmada Nicotiana rustica L.’nin su ve methanol ektresinin, PON 1 ve AChE enzim aktivitesi üzerine etkisine bakılmış. Nicotiana rustica Linn‘in su ektresi ceviz ile karıştırılmış maraş otu, (MPWW) ve methanol ektresi meşe ile karıştırılmış maraş otu (MPNR), ceviz ile karıştırılmış maraş otu (MPWW) ve bakır kazanlarında nemlendirilmiş maraş otu (MPCC) AChE üzerine inhibisyon etkisi (IC50 değerleri 0.104-0.203 mg ml -1 aralığında) gösterirken su ektresi MPCC ve methanol ektresi MPNR, MPWW ve MPCC ise PON 1 üzerine inhibisyon etkisi göstermiştir (IC50 değerleri 0.013-0. 25 mg ml -1 ). En yüksek radikal giderme aktivitesini MPNR su ektresi % 26 oranında DPPH radikal giderme aktivitesi gösterirken, MPCC su ektresi % 13 oranında ABTS radikal giderme aktivitesi göstermiştir., Being widely used since its discovery, tobacco contains many chemical substances including nitric oxide, hydrocarbons, aldehydes, phenols, quinone and semiquinone radicals, and causes the formation of oxygen-induced free radicals directly or indirectly. Therefore, it is important to know the oxidant/antioxidant properties, phenolic content of Nicotiana rustica Linn., and the effect studies on Paraoxonase (PON 1) and Acetylcholinesterase (AChE) which are important metabolic enzymes. In the study, the effect of extracts (water and methanol) of Nicotiana rustica Linn on PON 1 and AChE enzyme activity was examined. Water extract of Nicotiana rustica Linn Maras Powder with walnut, (MPWW) and methanol extract Maras Powder with Nicotiana rustica, (MPNR), Maras Powder with walnut, (MPWW) and Maras Powder with copper cauldron, (MPCC) had an inhibitory effect on AChE (IC50 values in the range of 0.104-0.203 mg ml-1). Also water extract MPCC and methanol extract MPNR, MPWW and MPCC showed an inhibition effect on PON 1 (IC50 values 0.013-0.225 mg ml -1). MPNR water extract showed the highest activity with 26% DPPH radical scavenging activity. MPCC water extract showed a 13% ABTS radical scavenging activity.

Graphical abstract, Highlights • Toombak, a form of moist smokeless tobacco from Sudan is placed as a dip in the oral cavity most commonly used by males. • The microbiome of Toombak predominantly consists of the phyla, Firmicutes and Actinobacteria while abundant species include Corynebacterium casei, Atopostipes suicloacalis and Oceanobacillus chironomi. • High concentrations of iron, volatile aldehydes and tobacco specific nitrosamines were found in Toombak and can lead to toxicity. • Toombak has a non-homogenous abrasive surface with a high sodium level in the ready to buy form that can damage the oral mucosa. • New measures must be taken in Sudan to limit harmful compounds in Toombak., Toombak is a smokeless tobacco produced from the Nicotiana rustica tobacco plant from Sudan. Pre-prepared and ready to buy Toombak samples were analysed using mass spectrometry (heavy metals), gas and liquid chromatography (metabolomics), 16S rRNA metagenomic sequencing (microbiome) and Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt), scanning electron microscopy with energy dispersive X-ray spectroscopy (SEM-EDX) and pH analysis. Chromium, cobalt, and copper were high in the pre-prepared form of Toombak while iron, tobacco specific nitrosamines (TSNAs), formaldehyde and acetaldehyde were high in both types. Firmicutes and Actinobacteria dominated Toombak. Samples of ready to buy Toombak showed inter-variational differences depending on place of purchase. We found Virgibacillus were increased in the pre-prepared form while Corynebacterium casei, Atopococus tabaci, Atopostipes suicloacalis, Oceanobacillus chironomi and Staphylococcus gallinarum were the most abundant species in the ready to buy forms. PICRUSt analysis highlighted increased activity of metal transport systems in the ready to buy samples as well as an antibiotic transport system. SEM-EDX highlighted large non-homogenous, irregular particles with increased sodium, while pH of samples was in the alkaline range. The final composition of Toombak is affected by its method of preparation and the end product has the potential to impart many negative consequences on the health of its users. TSNA levels observed in Toombak were some of the highest in the world while the micro-environment of Toombak supports a distinct microbiota profile.

The tobacco Nicotiana rustica is widely used as a trap crop in the fields of Nicotiana tabacum in China, by attracting oviposition of Helicoverpa assulta females, thus preventing damage to N. tabacum. The mechanism underlying the differential oviposition rates of H. assulta across these two tobacco species, however, is largely unknown. We investigated the mechanism of host plant acceptance of H. assulta with respect to these two tobaccos by using a two-choice behavioral bioassay and GC–MS. Our results indicate that both the leaves and inflorescences of N. rustica attracted significantly more eggs than the corresponding parts of N. tabacum. Extracts of leaves and inflorescences of N. rustica with two different solvents elicited similar oviposition patterns to the corresponding parts of the plants. Chemical analysis by GC–MS revealed that the volatiles of N. rustica contain larger amounts of nicotine than those of N. tabacum at the flowering stage. In addition, γ-terpinolene and β-elemene are found only in extracts of N. rustica. A two-choice bioassay on the individual compounds showed that γ-terpinolene, which is specific to the vegetative stage of N. rustica, and nicotine attracted oviposition by H. assulta. The volatile β-elemene, which is present only in N. rustica, was also attractive. We conclude that the larger amount of nicotine, and the species-specific γ-terpinolene and β-elemene may mediate the different oviposition rates of H. assulta females across N. rustica and N. tabacum.

It is known that tomato is one of the most susceptible vegetable crops affected by viral pathogens. Recently, Pepino mosaic virus, is widespread in greenhouse plantations in Europe the harmfulness of which leads to yield and marketability of products decrease. The analysis of tomato hybrids grown in greenhouses of the republic has shown that in different years of research, Pepino mosaic virus has been present both in mono infection and in combination with the other viruses from the genus Cucumovirus, Tobamovirus, Nepovirus and Tobravirus. Laboratory data indicate the susceptibility and ability to high accumulation of viral particles when test plants Datura stramonium L. and Nicotiana rustica L. have been infected with the pepino mosaic virus. It is determined that Aphis gossypii Glov., Frankliniella occidentalis Perg. and Trialeurodes vaporariorum Wetw. can participate in the vector transmission of the pathogen between host plants.

Promoters are key elements regulating gene expression levels, therefore their selection is an important step in genetic engineering research. The reporter gene gfp, which encodes green fluorescent protein (GFP), was transiently expressed in leaf tissues of Aztec tobacco Nicotiana rustica L. Compared to other species of the Nicotiana genus, Aztec tobacco has a large potential for expression of heterologous proteins, a large vegetative biomass, can be easily infiltrated, and is unpretentious in cultivation. Six genetic constructs were used with different promoter sequences: the 35S promoter of Cauliflower Mosaic Virus (35S CaMV), the double-enhanced 35S promoter (D35S CaMV), promoters of the RbcS1B and RbcS2B genes encoding the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) isolated from Arabidopsis thaliana (L.) Heynh., and promoters of the LHB1B1 and LHB1B2 genes from A. thaliana encoding chlorophyll a/b binding proteins. The gfp gene expression was detected visually, spectrofluorimetrically, and by protein content (Bradford assay) on the seventh day after infiltration. The highest level of expression was observed using the double-enhanced 35S promoter (D35S CaMV) and the lowest using the LHB1B1 gene promoter.

Two new pentenyl coumarins (1 and 2), together with three known pentenyl coumarins (3–5), were isolated from the roots and stems of Nicotiana rustica. Their structures were elucidated by spectroscopic methods, including extensive 1D and 2D NMR techniques. Compounds 1–5 were tested for their anti-rotavirus activity. The results revealed that compounds 1–5 exhibited potent anti-rotavirus activity with therapeutic index (TI) value in the range of 11.1–15.7.

Aim. Genetic constructs creation for studying the influence effect of the viral posttranscriptional silencing protein suppressor p19 on transient reporter green fluorescent protein (GFP) expression and accumulation. Methods. The Golden Gate molecular cloning method was used to create the genetic constructs; the leafy tissues of the Aztec tobacco plants (Nicotiana rustica L.) were infiltrated with a suspension of Agrobacterium tumefaciens L.; the gfp gene expression level was determined by spectrofluorometric and quantitative protein (Bradford method) assays. Results. As a result of the work, the pSPV2324 genetic construct was created, which contained the reporter gene for the green fluorescent protein gfp and the gene for the synthesis of the viral posttranscriptional silencing protein suppressor p19 and its effect on the accumulation of the recombinant GFP protein was determined. A comparative analysis of the gfp gene expression level without and with the suppressor protein synthesis gene in the genetic vector showed that the fluorescence level of GFP protein in Aztec tobacco tissues was 1.3 times higher during spectrofluorimetric analysis using the p19 suppressor gene construct. Conclusions. The positive effect of the viral suppressor silencing P19 gene on the accumulation of recombinant GFP protein in tissues plants of N. rustica L. was shown for the first time. The increase in GFP protein fluorescence when using the p19 suppressor protein construct in spectrofluorimetric analysis coincides with an increase in the total concentration of total water-soluble proteins and the level fluorescence of GFP protein in their native electrophoretic separation. Keywords: cloning, genetic constructs, transient expression, silencing protein suppressor p19, green fluorescent protein (GFP).

Cadmium (Cd) is known as one of the most hazardous elements in the environment and a persistent soil constraint toxic to all flora and fauna. In this study, we conducted physiological, biochemical, and transcriptomic analyses of Nicotiana rustica (N. rustica) and Nicotiana tabacum (N. tabacum) treated with CdCl2 to know the underlying molecular mechanisms of Cd accumulation. As a result, N. rustica had more dry weight than N. tabacum. Additionally, N. rustica accumulated higher Cd concentration (69.65 times), Cd2+ influx (1.32-fold), glutathione S-transferases (GST) enzyme activity (2.54 times), GSH/GSSG (oxidized form of GSH) ratio, increase of superoxide dismutase and CAT and a lower H2 O2 and superoxide (O2 •- ) accumulation in their roots than N. tabacum. Cd mainly distributed in the cytoplasm of both species and N. rustica had a significant proportion in the cell wall. Furthermore, the transcriptomic analysis revealed 173 and 710 differentially expressed genes (DEGs) between control and Cd-stressed plants in the leaves and roots of N. rustica, while 576 and 1543 DEGs were found in the leaves and roots of N. tabacum, respectively. In N. rustica, phenylpropanoid biosynthesis and phenylalanine metabolism were the most enriched pathways, while GSH metabolism, ATP-binding cassette transporters and phenylpropanoid biosynthesis were the most enriched in N. tabacum. Finally, we found that DEGs related to metal influx, sequestration, remobilization, and chelation were responsible for Cd accumulation. These results indicated that N. rustica accumulated higher Cd content than N. tabacum, suggesting that each species utilized different response mechanism under the same Cd stress conditions. The DEGs identified in this study might lead to the identification of genes or pathways related to Cd regulation. This study identifies important regulators related to Cd accumulation.

Nous avons défini le complexe tabagique selon quatre champs applicables : les pipes à fumer, le tabac, le rôle du tabac dans les rapports Autochtones-Européens et dans les échanges. Le but de ce mémoire est de mieux saisir ce que les pipes à fumer de la période historique nous dévoilent dans le contexte de l’archipel montréalais. Le dépouillement des rapports de fouille a permis de retirer d’innombrables fragments de pipes à fumer que nous retrouvons dans les contextes archéologiques de l’archipel montréalais pour la période 1642−1760. Les récits d’époque nous informent sur les habitudes reliées à la consommation du tabac ainsi que les rôles du tabac et des pipes à fumer dans les relations entre Autochtones et Européens et des processus d’échange. Des recherches sur le tabac sont venues ajouter des éléments cruciaux à ces deux sources d’information. Cette étude a été limitée à l’archipel montréalais, plus une aire de 10 km autour de celui-ci. La période étudiée est de 1642 à 1760. Nous avons constaté que les fragments des pipes à fumer se retrouvent majoritairement dans les zones de contact démontrant ainsi l’importance de ces objets dans les échanges. Ces zones de contact sont les endroits où le métissage prend place. Le tabac, que nous ne pouvons dissocier des pipes à fumer, joue aussi un rôle majeur dans les relations amérindiennes-européennes. Des dons de Nicotiana tabacum ont permis de solidifier des liens de confiance primordiaux entre les deux groupes dans les processus d’échange. Le tabac et les pipes à fumer, ont aussi subit le processus de transfert culturel, mais, le degré varie selon le type de pipes à fumer., We defined the smoking complex according to four applicable fields: smoking pipes, tobacco, Indigenous -European relations and the role of tobacco in trade. The purpose of this research is to better understand what smoking pipes reveal to us in the context of the Montreal archipelago. The review of the excavation reports allowed us to retrieve information regarding the innumerable fragments of smoking pipes that we find in the archaeological contexts of the Montreal archipelago for the period 1642−1760. The ethnohistorical publications inform us about the habits and customs related to the use of tobacco as well as the roles that tobacco and smoking pipes played in Indigenous-European relations and exchanges. Tobacco studies have also added crucial elements to these two sources of information. This study was limited to the Montreal archipelago, plus an area of 10 km around it. The study period is from 1642 to 1760. We found that the fragments of smoking pipes are mostly found in contact zones demonstrating the cultural importance of these objects. Nicotiana tabacum strengthened bonds of trust between the two groups. These contact zones are the places where métissage takes place. Inseparable from smoking pipes, tobacco also played a major role in Amerindian-European relations. Gifts of Nicotiana tabacum favoured consolidation between the two groups and built the primordial trust necessary in exchange processes. Tobacco and smoking pipes are also part of the process of cultural transfer, but to a variable degree according to the type of smoking pipes.

The genus Nicotiana of the family Solanaceae, commonly referred to as tobacco plants, are a group cultivated as garden ornamentals. Besides their use in the worldwide production of tobacco leaves, they are also used as evolutionary model systems due to their complex development history, which is tangled by polyploidy and hybridization. Here, we assembled the plastid genomes of five tobacco species, namely N. knightiana, N. rustica, N. paniculata, N. obtusifolia and N. glauca. De novo assembled tobacco plastid genomes showed typical quadripartite structure, consisting of a pair of inverted repeats (IR) regions (25,323–25,369 bp each) separated by a large single copy (LSC) region (86,510 –86,716 bp) and a small single copy (SSC) region (18,441–18,555 bp). Comparative analyses of Nicotiana plastid genomes showed similar GC content, gene content, codon usage, simple sequence repeats, oligonucleotide repeats, RNA editing sites and substitutions with currently available Solanaceae genomes sequences. We identified twenty highly polymorphic regions mostly belonging to intergenic spacer regions (IGS), which could be appropriate for the development of robust and cost-effective markers to infer the phylogeny of genus Nicotiana and family Solanaceae. Our comparative plastid genome analysis revealed that the maternal parent of the tetraploid N. rustica was the common ancestor of N. paniculata and N. knightiana, and the later species is more closely related to N. rustica. The relaxed molecular clock analyses estimated that the speciation event between N. rustica and knightiana appeared 0.56 Ma (HPD 0.65–0.46). The biogeographical analysis showed a south-to-north range expansion and diversification for N. rustica and related species, where N. undulata and N. paniculata evolved in North/Central Peru, while N. rustica developed in Southern Peru and separated from N. knightiana, which adapted to the Southern coastal climatic regimes. We further inspected selective pressure on protein-coding genes among tobacco species to determine if this adaptation process affected the evolution of plastid genes. These analyses indicated that four genes involved in different plastid functions, such as DNA replication (rpoA) and photosynthesis (atpB, ndhD and ndhF), came under positive selective pressure as a result of specific environmental conditions. Genetic mutations of the following genes might have contributed to the survival and better adaptation during the evolutionary history of tobacco species.

Residues from ancient artifacts can help identify which plant species were used for their psychoactive properties, providing important information regarding the deep-time co-evolutionary relationship between plants and humans. However, relying on the presence or absence of one or several biomarkers has limited the ability to confidently connect residues to particular plants. We describe a comprehensive metabolomics-based approach that can distinguish closely related species and provide greater confidence in species use determinations. An ~1430-year-old pipe from central Washington State not only contained nicotine, but also had strong evidence for the smoking of Nicotiana quadrivalvis and Rhus glabra, as opposed to several other species in this pre-contact pipe. Analysis of a post-contact pipe suggested use of different plants, including the introduced trade tobacco, Nicotiana rustica. Ancient residue metabolomics provides a new frontier in archaeo-chemistry, with greater precision to investigate the evolution of drug use and similar plant-human co-evolutionary dynamics.

Cadmium (Cd) is a severely toxic and carcinogenic heavy metal. Cigarette smoking is one of the major source of Cd exposure in humans. Nicotiana tabacum is primarily a leaf Cd accumulator, while Nicotiana rustica is a root Cd accumulator among Nicotiana species. However, little is known about the mechanisms of differential Cd translocation and accumulation in Nicotiana. To find the key factors, Cd concentration, Cd chemical forms, and transcriptome analysis were comparatively studied between N. tabacum and N. rustica under control or 10 μM Cd stress. The leaf/root Cd concentration ratio of N. tabacum was 2.26 and that of N. rustica was 0.14. The Cd concentration in xylem sap of N. tabacum was significantly higher than that of N. rustica. The root of N. tabacum had obviously higher proportion of ethanol extractable Cd (40%) and water extractable Cd (16%) than those of N. rustica (16% and 6%). Meanwhile the proportion of sodium chloride extracted Cd in N. rustica (71%) was significantly higher than that in N. tabacum (30%). A total of 30710 genes expressed differentially between the two species at control, while this value was 30,294 under Cd stress, among which 27,018 were collective genes, manifesting the two species existed enormous genetic differences. KEGG pathway analysis showed the phenylpropanoid biosynthesis pathway was overrepresented between the two species under Cd stress. Several genes associated with pectin methylesterase, suberin and lignin synthesis, and heavy metal transport were discovered to be differential expressed genes between two species. The results suggested that the higher accumulation of Cd in the leaf of N. tabacum depends on a comprehensive coordination of Cd transport, including less cell wall binding, weaker impediment by the Casparian strip, and efficient xylem loading.

Nicotiana rustica L. (Aztec tobacco) is the only Nicotiana species, except common tobacco ( N. tabacum L.), which is cultivated for tabacco products. The leaves of N. rustica , however, accumulate various specialized metabolites of potential interest. Therefore, the objective of this study was to evaluate certain classes of metabolites (by HPLC and GC-MS) in the leaves, the essential oil (EO), concrete and resinoid of N. rustica . Three pentacyclic triterpenes were identified in the leaves (by HPLC): betulin (252.78 µg g -1 ), betulinic (182.53 µg g -1 ) and oleanolic (69.44 µg g -1 ) acids. The dominant free phenolic acids in the leaves (by HPLC) were rosmarinic (4257.38 µg g -1 ) and chlorogenic (1714.40 µg g -1 ), and conjugated forms of vanillic (3445.71 µg g -1 ), sinapic (1963.11 µg g -1 ), and syringic (1784.96 µg g -1 ). The major flavonoids in the leaves were luteolin (960.44 µg g -1 ), apigenin (880.66 µg g -1 ) and hyperosid (780.72 µg g -1 ). The GS-MS profiling of the EO identified 19 components and the major ones were phytol (43.68 %) , solanone (5.54 % ) , cis-5- b utyl-4-methyldihydrofuran-2(3H)-one (5 . 23 % ) , dihydro-β-ionone (4.25 % ) , α-ionene (3 . 54 % ) , and β-damascenone ( 3. 03 % ) . The major volatiles in the concrete were isoamyl alcohol (28 . 82 %), oxynicotine ( 9 . 02 %), phytol ( 7.80 %), 4-mеthyl-1-penthanol ( 6 . 33 %), cotinine ( 5.55 %) and 3-metyl-3-penthanol ( 4.09 %). Resinoid composition was dominant by nicotine ( 39 . 75 % ), phytol ( 11 . 23 %), eicosane ( 4 . 88 %), diethyl phthalate ( 4 . 19 %), dibutyl phthalate ( 3 . 48 %) and solanone ( 3 . 27 %) . Concrete and resinoid showed weak antibacterial activity . These results create grounds for considering N. rustica as a source to obtain aroma or other bioproducts.

Objective This study aimed to investigate the effects of Maras powder (a type of smokeless tobacco obtained from Nicotiana rustica Linn and mixed with the ashes of wood, especially from oak, walnut or grapevine) on the microRNA (miRNA) deregulation of oral mucosa, and it compares these effects with those of smoking. Methodology Oral mucosal samples were collected from 74 patients, consisting of 16 nonusers, 26 smokers, and 32 Maras powder users. Genes associated with oral cancer were selected and 90 microRNAs targeting these genes were identified. MicroRNA were isolated and purified using the microRNA isolation kit. MicroRNA were expressed using Fluidigm RT-PCR. Results A positive correlation between the duration of Maras powder use with miR-31 expression levels, and a negative correlation between the Maras powder chewing time and miR-372 expression levels was found. In addition, there is a negative correlation between the amount of Maras powder consumed and expression levels of miR-375, miR-378a, miR-145, and miR-10b; moreover, another negative correlation is observed between the number of cigarettes consumed and the expression levels of miR-23a, miR-23b, miR-203a, miR-200b, and miR-375. However, miR-200b and miR-92a levels were downregulated significantly more in Maras powder users when compared with smokers and nonusers (p

Traditional tobacco was originally considered to be sacred by Native Americans and was used for ceremonial and medicinal purposes for thousands of years. It was, in fact, what we know today as the Nicotiana rustica species that was considered to be “traditional” tobacco. In addition to its traditional purposes, before the arrival of Europeans, Native Americans consumed tobacco by chewing it, in a very similar manner as coca leaves. The chewing custom was gradually adopted by the new settlers, resulting in commercial production and spreading across multiple continents over time. Another indigenous way of consuming tobacco, sniffing pulverized leaves, was first brought to Europe by the Spanish in the early sixteenth century and also became popular among Europeans. The most popular method of tobacco consumption, however, is smoking cigarettes. Although it was likely nicotine that was the primary driver behind the popularity of tobacco products, there is an extremely complex biosynthetic mechanism that leads to the production of many different compounds in the tobacco plant. Without a doubt, plants are one of the most important sources of valuable compounds. Our food, paper, clothes, and numerous medicines are derived from them. Even the fossil fuels we use so much today originate from the decomposition of natural plant biomass. Common tobacco (Nicotiana tabacum L.), though primarily associated today with consumer goods production, is no different from other plants and efficiently produces variety of valuable organic compounds. This chapter describes tobacco biosynthetic potential, from alkaloids and phenolics to complex cembranoids, and presents in detail state-of-the-art knowledge of metabolic pathways as well as the methods and factors affecting their accumulation.

The present study aimed to investigate the effects of ethanolamine on glycine betaine and proline metabolism in Nicotiana rustica under salt stress. The in vitro grown tobacco (Nicotiana rustica) plants were pretreated with ethanolamine (at concentrations 70, 130, 270, and 530 μM for biochemical analysis and only at the concentration of 530 μM for molecular analysis) and then transferred to Murashige and Skoog medium containing 200 mM NaCl for 3 weeks. Our results showed that ethanolamine promoted glycine betaine biosynthesis by an increase in betaine aldehyde dehydrogenase (BADH) gene expression and BADH enzymatic activity. Moreover, ethanolamine pretreatment possibly reduced proline content in salt stressed plants via its negative effect on Δ-pyrroline-5-carboxylate synthase (P5CS) gene expression and P5CS enzymatic activity and its positive effect on proline dehydrogenase (PDH) gene expression and PDH activity.

Effect of potassium (K) application through leaves (LA) or roots (RA) was studied in tobacco plants grown under K deficiency and drought stress conditions. Application of K was effective in improving the shoot growth only under drought conditions, whereas root biomass and length responded under both watering regimes. Under drought conditions, photosynthesis and transpiration activities increased upon K application leading to a reduced water use efficiency. Both RA and LA increased the leaf water potential, relative water content and turgor under both well-watered and drought conditions; RA was more effective than LA in the recovery of leaf turgor. Analyses of water relation parameters in different aged leaves showed lower susceptibility of the middle-aged leaves to both K deficiency and drought stresses than the upper and lower leaves; this phenomenon was accompanied by a more conservative control of water loss in the middle-aged leaves. In contrast, proline was accumulated in the young leaves, and K application increased it further. Although various organic osmolytes were accumulated under the combinative effect of K deficiency and drought stress, they did not exceed the amounts found in the control (well-watered +K) plants and were merely a result of the concentration effect. Collectively, our results revealed that the majority of leaf biochemical responses to drought stress are developmentally regulated processes. In addition, the alleviating effect of both RA and LA despite higher water loss indicated that an improved stomatal function upon K application allowed carbohydrates synthesis, thus, enhancing plant growth under water stress.

Tomato spotted wilt orthotospovirus (TSWV) was first reported in 2004 from paprika in South Korea (Kim et al., 2004), where it is currently widespread. TSWV infections were reported in chili pepper, tomato, weeds, and ornamental plant species in South Korea (Choi et al., 2014; Choi and Choi, 2015; Yoon et al., 2016; Yoon et al., 2018; Yoon et al., 2019). One of the best strategies for TSWV management is planting resistant cultivars containing the Tsw gene. In 2019 virus-like symptoms were observed in chili pepper (Capsicum annuum) plants bearing the Tsw gene in Anseong-si, South Korea. The infected chili peppers showed mosaic and wilting followed by necrosis on leaves and fruits in the field. To identify the causal virus, symptomatic leaf samples were analyzed using ImmunoStrip kits (Agdia, USA); we detected three pepper-infecting viruses: Pepper mild mottle virus, Cucumber mosaic virus, and TSWV. TSWV was only detected from 40 naturally infected chili pepper plants exhibiting virus-like symptoms. To further confirm the presence of TSWV (named TSWV-P1), we amplified reverse-transcription polymerase chain reaction (RT-PCR) products for L, M, and S RNA segments using tospovirus-specific and TSWV-specific primers (Batuman et al., 2014). Expected fragments of 445, 868, and 777 bp in length were amplified and sequenced. The complete genome sequences of TSWV-P1 from a symptomatic chili pepper plant were also determined using TSWV-specific primers (Choi et al., 2014; Lian et al., 2013). The complete genome sequences of TSWV-P1 were deposited to GenBank (LC549179, LC549180, and LC549181). The sequences of each fragment were identical to a consensus sequence, showing 99.1%, 98.5%, and 98.6% identity with TSWV-L, M, and S RNA (KP008132, AY744492, and KP008134), respectively. These results clearly showed only a single TSWV infection among the naturally infected chili pepper plants, without reassortment between TSWV and another tospovirus. To confirm whether TSWV-P1 is a resistance-breaking (RB) strain, Nicotiana rustica was mechanically inoculated with sap from leaves of the infected pepper samples to propagate TSWV-P1. A non-RB TSWV isolate (TSWV-Kor-lisianthus) from lisianthus was used as a control (Yoon et al., 2017). Two resistant (with Tsw) and two susceptible chili pepper cultivars (20 plants per cultivar) were mechanically inoculated with sap from leaves of the TSWV-infected N. rustica. The incidence rates of disease caused by TSWV-P1 were 90-100% for resistant and 95-100% for susceptible cultivars. In contrast, TSWV-Kor-lisianthus caused symptoms only in the susceptible pepper cultivars (90-100% incidence). TSWV infection in representative plants was confirmed using the TSWV- ImmunoStrip kit and RT-PCR. The NSs gene of TSWV-P1 consists of 1,404 nucleotides (468 amino acids); sequence analysis of the TSWV-P1 NSs gene showed high nucleotide (99.7%) and amino acid identities (99.8%) with the NSs sequences of two TSWV isolates (FR693035, CBX24121). Protein sequence analysis of TSWV-P1 NSs revealed that no amino acid mutation was associated with those of a representative TSWV RB strain, as previously described (Almasi et al., 2017), suggesting that TSWV-P1 is a RB strain. Because this TSWV-P1 can overcome resistance conferred by the Tsw gene in commercially grown chili pepper cultivars, it represents a potential threat to pepper production in South Korea.