Non-targeted discovery of high-value bio-products in Nicotiana glauca L: a potential renewable plant feedstock.

The evaluation of plant-based feedstocks is an important aspect of biorefining. Nicotiana glauca is a solanaceous, non-food crop that produces large amounts of biomass and is well adapted to grow in suboptimal conditions. In the present article, compatible sequential solvent extractions were applied to N. glauca leaves to enable the generation of enriched extracts containing higher metabolite content comparing to direct leaf extracts. Typically, between 60 to 100 metabolite components were identified within the fractions. The occurrence of plant fatty acids, fatty acid alcohols, alkanes, sterols and terpenoids was detected by gas liquid chromatography-mass spectrometry (GC-MS) and metabolite identification was confirmed by comparison of physico-chemical properties displayed by available authentic standards. Collectively, co-products such waxes, oils, fermentable sugars, and terpenoids were all identified and quantified. The enriched fractions of N. glauca revealed a high level of readily extractable hydrocarbons, oils and high value co-products. In addition, the saccharification yield and cell wall composition analyses in the stems revealed the potential of the residue material as a promising lignocellulosic substrate for the production of fermentable sugars. In conclusion a multifractional cascade for valuable compounds/commodities has been development, that uses N. glauca biomass. These data have enabled the evaluation of N. glauca material as a potential feedstock for biorefining.

Fatty alcohols, a minor component of the tree tobacco surface wax, are associated with defence against caterpillar herbivory.

Despite decades of research resulting in a comprehensive understanding of epicuticular wax metabolism, the function of these almost ubiquitous metabolites in plant-herbivore interactions remains unresolved. In this study, we examined the effects of CRISPR-induced knockout mutations in four Nicotiana glauca (tree tobacco) wax metabolism genes. These mutations cause a wide range of changes in epicuticular wax composition, leading to altered interactions with insects and snails. Three interaction classes were examined: chewing herbivory by seven caterpillars and one snail species, phloem feeding by Myzus persicae (green peach aphid) and oviposition by Bemisia tabaci (whitefly). Although total wax load and alkane abundance did not affect caterpillar growth, a correlation across species, showed that fatty alcohols, a minor component of N. glauca surface waxes, negatively affected the growth of both a generalist caterpillar (Spodoptera littoralis) and a tobacco-feeding specialist (Manduca sexta). This negative correlation was overshadowed by the stronger effect of anabasine, a nicotine isomer, and was apparent when fatty alcohols were added to an artificial lepidopteran diet. By contrast, snails fed more on waxy leaves. Aphid reproduction and feeding activity were unaffected by wax composition but were potentially affected by altered cutin composition. Wax crystal morphology could explain the preference of B. tabaci to lay eggs on waxy wild-type plants relative to both alkane and fatty alcohol-deficient mutants. Together, our results suggest that the varied responses among herbivore classes and species are likely to be a consequence of the co-evolution that shaped the specific effects of different surface wax components in plant-herbivore interactions.

Tree tobacco (Nicotiana glauca) cuticular wax composition is essential for leaf retention during drought, facilitating a speedy recovery following rewatering.

Despite decades of extensive study, the role of cuticular lipids in sustaining plant fitness is far from being understood. We utilized genome-edited tree tobacco (Nicotiana glauca) to investigate the significance of different classes of epicuticular wax in abiotic stress such as cuticular water loss, drought, and light response. We generated mutants displaying a range of wax compositions. Four wax mutants and one cutin mutant were extensively investigated for alterations in their response to abiotic factors. Although the mutations led to elevated cuticular water loss, the wax mutants did not display elevated transpiration or reduced growth under nonstressed conditions. However, under drought, plants lacking alkanes were unable to reduce their transpiration, leading to leaf death, impaired recovery, and stem cracking. By contrast, plants deficient in fatty alcohols exhibited elevated drought tolerance, which was part of a larger trend of plant phenotypes not clustering by a glossy/glaucous appearance in the parameters examined in this study. We conclude that although alkanes have little effect on whole N. glauca transpiration and biomass gain under normal, nonstressed conditions, they are essential during drought responses, since they enable plants to seal their cuticle upon stomatal closure, thereby reducing leaf death and facilitating a speedy recovery.

The Biosynthesis of Non-Endogenous Apocarotenoids in Transgenic Nicotiana glauca.

Crocins are high-value compounds with industrial and food applications. Saffron is currently the main source of these soluble pigments, but its high market price hinders its use by sectors, such as pharmaceutics. Enzymes involved in the production of these compounds have been identified in saffron, Buddleja, and gardenia. In this study, the enzyme from Buddleja, BdCCD4.1, was constitutively expressed in Nicotiana glauca, a tobacco species with carotenoid-pigmented petals. The transgenic lines produced significant levels of crocins in their leaves and petals. However, the accumulation of crocins was, in general, higher in the leaves than in the petals, reaching almost 302 µg/g DW. The production of crocins was associated with decreased levels of endogenous carotenoids, mainly ß-carotene. The stability of crocins in leaf and petal tissues was evaluated after three years of storage, showing an average reduction of 58.06 ± 2.20% in the petals, and 78.37 ± 5.08% in the leaves. This study illustrates the use of BdCCD4.1 as an effective tool for crocin production in N. glauca and how the tissue has an important impact on the stability of produced high-value metabolites during storage.

UPLC-HRESI-MS and GC-MS analysis of the leaves of Nicotiana glauca.

The alkaloid-rich fraction obtained by fractionation of the crude methanolic extract of the leaves of wild tobacco tree Nicotiana glauca Graham (Solanaceae) was analyzed using UPLC-MS and GC-MS. Anabasine, a piperidine alkaloid, was identified as the major constituent with approximately 60 % (m/m) of the alkaloid-rich fraction. In addition to anabasine, six secondary metabolites were identified using high-resolution UPLC-MS. Anabasine was quantified in the leaves to be 1 mg g-1 dry plant material. The GC-MS analysis revealed five compounds with anabasine as the major component, while nicotine was not detected. Moreover, GC-MS was used for the analysis of the volatile oil that was obtained by hydro-distillation from the leaves of N. glauca. The volatile plant oil was found to be rich in oxygenated sesquiterpenes (e.g., ß-bisabolol) and carboxylic acids and esters (e.g., ethyl linoleate and hexadecanoic acid), whereas anabasine was not detected.

Intoxicación severa por Nicotiana glauca (Palán Palán) con miocardio atontado / Severe poisoning by Nicotiana glauca (Palán Palán) with stunned myocardium

Resumen Nicotiana glauca también llamada Palán Palán, es un arbusto con hojas verdes azuladas y despulidas y una flor amarilla tubular pendulante que presenta alcaloides piridínicos, como nicotina, nornicotina, anatabina y anabastina (análogo estructural de la Nicotina). Se presenta el caso clínico de una paciente de 50 años con cuadro agudo de debilidad muscular generalizada que evoluciona con paro respiratorio, tras la ingesta accidental de una cantidad desconocida de hojas de Nicotiana glauca, cultivadas en una huerta hogareña mediante técnica de hidroponía y confundidas por su conviviente con espinaca. Presentó aumento de lactato y Troponina Ultra Sensible e Hipoquinesia Global de Ventrículo Izquierdo en el ecocardiograma, compatible con Aton tamiento Miocárdico (AM), que evolucionó favorablemente. Si bien hay pocos reportes, se han informado muertes de animales y humanos, tras la ingesta accidental de Nicotiana glauca. El inicio del cuadro es rápido, con patrón bifásico, con vómitos y estímulo simpático, seguido por bloqueo ganglionar y neuromuscular, pudiendo presentar paro respiratorio, shock, convulsiones y coma. El AM es una disfunción miocárdica prolongada con retorno gradual de la actividad contráctil, posterior a un episodio breve de isquemia grave, puede ser asintomático, pudiendo presentar alteraciones en el electrocardiograma, enzimas cardíacas o ecocardiograma. Generalmente presenta pronóstico favorable, pudiendo presentar insuficiencia cardíaca ante patologías concurrentes o aumento de requerimientos de oxígeno.

Abstract Nicotiana glauca is a shrub with bluish green leaves and a pendulous tubular yellow flower. It has pyridine alkaloids, such as nicotine, nornicotine, anatabine and anabastine (structural analog of Nicotine). We present the case of a 50 years old pa- tient with acute generalized muscle weakness that evolves to respiratory arrest, after accidentally ingesting an unknown quantity of Nicotiana glauca leaves, grown in a home vegetable garden, using a hydroponic technique and confused by her cohabiting with spinach. She presented increased lactate and Ultra Sensitive Troponin and Left Ventricular Global Hypokinesia in the echo- cardiogram, compatible with Myocardial Stunned, that it evolved favorably. There are few reports, animal and human deaths have been reported following accidental ingestion of Nicotiana glauca. The onset of the symptoms is early, with a biphasic pattern, with vomiting and sympathetic stimulation, followed by ganglionic and neuromuscular blockage and may present respiratory arrest, shock, seizures and coma. Myocardial Stunned is a prolonged myocardial dysfunction with gradual return of contractile activity after a brief episode of severe ischemia, it can be asymptomatic, and it can present alterations in the electrocardiogram, cardiac enzymes or echocardiogram. Generally presents a benign prognosis, being able to present heart failure with concurrent patholo- gies or increased requirements.

Pseudomonas palmensis sp. nov., a Novel Bacterium Isolated From Nicotiana glauca Microbiome: Draft Genome Analysis and Biological Potential for Agriculture.

A novel Pseudomonas, designated strain BBB001T, an aerobic, rod-shaped bacterium, was isolated from the rhizosphere of Nicotiana glauca in Las Palmas Gran Canaria, Spain. Genomic analysis revealed that it could not be assigned to any known species of Pseudomonas, so the name Pseudomonas palmensis sp. nov. was proposed. A 16S rRNA gene phylogenetic analysis suggested affiliation to the Pseudomonas brassicae group, being P. brassicae MAFF212427 T the closest related type strain. Upon genomic comparisons of both strains, all values were below thresholds established for differentiation: average nucleotide identity (ANI, 88.29%), average amino acid identity (AAI, 84.53%), digital DNA-DNA hybridization (dDDH, 35.4%), and TETRA values (0.98). When comparing complete genomes, a total of 96 genes present exclusively in BBB001T were identified, 80 of which appear associated with specific subsystems. Phenotypic analysis has shown its ability to assimilate glucose, potassium gluconate, capric acid malate, trisodium citrate, and phenylacetic acid; it was oxidase positive. It is able to produce auxins and siderophores in vitro; its metabolic profile based on BIOLOG Eco has shown a high catabolic capacity. The major fatty acids accounting for 81.17% of the total fatty acids were as follows: C16:0 (33.29%), summed feature 3 (22.80%) comprising C16:1 ω7c and C16:1 ω6c, summed feature 8 (13.66%) comprising C18:1 ω7c, and C18:1ω6c and C17:0 cyclo (11.42%). The ability of this strain to improve plant fitness was tested on tomato and olive trees, demonstrating a great potential for agriculture as it is able to trigger herbaceous and woody species. First, it was able to improve iron nutrition and growth on iron-starved tomatoes, demonstrating its nutrient mobilization capacity; this effect is related to its unique genes related to iron metabolism. Second, it increased olive and oil yield up to 30% on intensive olive orchards under water-limiting conditions, demonstrating its capacity to improve adaptation to adverse conditions. Results from genomic analysis together with differences in phenotypic features and chemotaxonomic analysis support the proposal of strain BBB001T (=LMG 31775T = NCTC 14418T) as the type strain of a novel species for which the name P. palmensis sp. nov is proposed.

Direct Evidence That Sunbirds' Gut Microbiota Degrades Floral Nectar's Toxic Alkaloids.

Orange-tufted sunbirds (Cinnyris osea) feed on the nectar of the tobacco tree (Nicotiana glauca) which contains toxic pyridine alkaloids characterized by high concentrations of anabasine and much lower concentrations of nicotine. We aimed at determining whether the gut microbiota of sunbirds harbors bacterial species that enable the birds to cope with these toxic alkaloids. An in vivo experiment that included 12 birds showed that inducing dysbiosis in sunbirds' guts by the addition of sulfamethoxazole and trimethoprim, significantly reduced the birds' ability to degrade anabasine (n = 3) compared to control birds (n = 3) with undisturbed microbiota. Sunbirds whose gut bacterial communities were altered by the antibacterial agents and who were fed with added nicotine, also showed a lower percentage of nicotine degradation (n = 3) in their excreta compared to the sunbirds with undisturbed microbiota (n = 3), though this difference was not significant. In an in vitro experiment, we studied the ability of Lactococcus lactis, Enterobacter hormaechei, Chryseobacterium gleum, Kocuria palustris, and Methylorubrum populi that were isolated from sunbirds' excreta, to degrade anabasine and nicotine. By using gas chromatography-mass spectrometry (GC-MS) analysis, we successfully demonstrated, for the first time, the ability of these species to degrade the focal secondary metabolites. Our findings demonstrate the role of gut bacteria in detoxifying toxic secondary metabolites found in the N. glauca nectar. The degradation products may supply the birds with nitrogen which is scarce in nectar-rich diets. These findings support another role of bacteria in mediating the interactions between plants and their pollinators.

Phytoconstituents screening and antimicrobial activity of the invasive species Nicotiana glauca collected from Al-Baha region of Saudi Arabia.

Nicotiana glauca belonging to the family Solanaceae is an exotic perinnial bushy species that recently has invaded the mountainous areas of Al-Baha region at the south-west of Saudi Arabia, causing degredation to soil and vegetation composition of those mountains. Accordingly, this plant must be either removed or exploited as a source of useful products such as medicines as it has been proven to contain many effective compounds with therapeutic properties. Thus, the current study aimd to screening N. glauca preliminary phytoconsitiuents and its anticmicrobial activity. Results showed that varying amounts of alkaloids, steroid, tannins, flavonoid, were present in the extracts of N. glauca leaves, stems and flowers. Saponins were present only in the flowers extract. However, all previuos bioactive compounds were absent in the roots except alkaloids, therefore, roots posses lesser amount of the phytoconstituents compared to the other parts of the plants. Leaves extracts inhibited the growth of E. coli (16.3 ± 0.71 mm) and also inhibited the growth of S. aureus (11.0 ± 0.23 mm), while, the flower extracts also inhibited the growth of E. coli (6.7 ± 0.65 mm), and also inhibited the growth of S. aureus (15.8 ± 0.52 mm). This result coincide with the previous results of the screening of N, glauca phytochemicals where the highest amounts found in leaves and flowers extracts. The inhibition zone of the antibiotic gentamycin (reference control) were 32 mm in S. aureus and 38 mm in E. coli. The inhibition zone of gentamycin (reference control) were 32 mm in S. aureus and 38 mm in E. coli. The minimum inhibitory concentration (MIC) of ethyl acetate was ranged (1.5-2 mg/ml). The lower MIC is an indication of high effectiveness of extract. Therefore, such bioactive property would support the idea that removing N. glauca from the mountainous areas of Al-Baha, Saudi Arabia, is not the only solution, but the plant can be exploited as a beneficial source of medicinal and economic purposes such as; antibacterial, antifungal and insecticidal, however, further comprehensive exploration is recommended to confirm such propreties.

Impact of the invasive plant species "Nicotiana glauca" toxins on the larvae of the invasive insect species "Rhynchophorus ferrugineus": A damaging pest of date palm trees in Saudi Arabia.

The wild tree tobacco (Nicotiana glauca) is an alien species that invaded vast areas of the Southwestern region of Saudi Arabia. While, the Red Palm Weevil (RPW) (Rhynchophorus ferrugineus) is considered to be the most damaging invasive insect species of palm trees all over the kingdom of Saudi Arabia, causing major economic losses to farmers and the economy of the country. Using conventional insecticides to control harmful insects such as RPW has undesirable effects on the environment and human health. Alternatively, using biocontrol agents such as poisonous extracts from N. glauca might be a better approach in pest management and can be considered as an eco-friendly, cost-effective, and safe alternative. Therefore, the current study aimed to evaluate the larvicidal effect of N. glauca aqueous extracts against the red palm weevil larvae. The plant specimens were collected from Al-Baha region in the Southwest of Saudi Arabia. Each single test consisted of 20 larvae, and N. glauca preparations were; 1, 1.5, 2, 2.5, and 3 ml, besides the control test. Results obtained for the effect of botanical extracts; leaf, flower, stem and root against R. ferrugineus larvae for an exposure period of 24 hr. at the concentrations of 2.8, 4.2, 6.0, 7.0 and 8.0 ppm. The concentrations for N. glauca extracts reflected an LC50 of 2.7 ppm for leave, 2.6 ppm for flower, 2.8 ppm for stem and 7.00 ppm for root. While, the same concentrations extracts reflected an LC95of 11 ppm for leaf, 9.6 ppm for flower, 8.9 ppm for stem and 13.00 ppm for root. These results showed that N. glauca extracts have a remarkable potentiality as insecticidal substances that can be used as an ecofriendly integrated approach for the management of R. ferrugineus.

Geographic partitioning or environmental selection: What governs the global distribution of bacterial communities inhabiting floral nectar?

Floral nectar harbors microbial communities which have significant impacts on its chemistry, volatiles, nutritional contents, and attractiveness for pollinators. Yet, fundamental knowledge regarding the structure and composition of nectar-associated microbiomes remains largely unknown. Especially elusive are the environmental factors and spatial effects that shape nectar-inhabiting microbial communities. The aim of this study was to explore and analyze the role of geographical and environmental factors affecting the composition and global distribution of floral nectar microbiota. We explored and compared the structure of bacterial communities inhabiting the floral nectar of the widely spread and invasive tobacco tree (Nicotiana glauca) in six continents: South and North America, Australia, Europe, Africa, and Asia, using 16S rRNA gene sequencing. Environmental abiotic data for each sampled plant was obtained from the Worldclim database and applied for inferring the effects of environmental conditions on bacterial community structure and diversity. Most abundant in the nectar were the Proteobacteria, Firmicutes, and Actinobacteria phyla, with Acinetobacter and Rosenbergiella (Proteobacteria) being the dominant bacterial genera that contributed most to the dissimilarities between sites. Acinetobacter and Rosenbergiella abundances were negatively correlated and significantly higher in the Mediterranean regions (Greece, Israel, and the Canary Islands) compared to Argentina and Australia. Temperature, site-elevation, rainfall, and density of vegetation were found to have significant effects on the structure and diversity of these bacterial communities in the nectar. Vegetation density was positively correlated with microbial diversity, while increased temperatures and elevation reduced the diversity and evenness of bacterial communities. Mantel's test showed that the similarity between the bacterial communities' composition significantly decreased as distances between them increased. We conclude that both geographical distance and local environmental abiotic conditions affect and shape the composition and diversity of nectar inhabiting bacterial communities.

An essay on ecosystem availability of Nicotiana glauca graham alkaloids: the honeybees case study.

BACKGROUND: Invasive plant species pose a significant threat for fragile isolated ecosystems, occupying space, and consuming scarce local resources. Recently though, an additional adverse effect was recognized in the form of its secondary metabolites entering the food chain. The present study is elaborating on this subject with a specific focus on the Nicotiana glauca Graham (Solanaceae) alkaloids and their occurrence and food chain penetrability in Mediterranean ecosystems. For this purpose, a targeted liquid chromatography electrospray tandem mass spectrometric (LC-ESI-MS/MS) analytical method, encompassing six alkaloids and one coumarin derivative, utilizing hydrophilic interaction chromatography (HILIC) was developed and validated. RESULTS: The method exhibited satisfactory recoveries, for all analytes, ranging from 75 to 93%, and acceptable repeatability and reproducibility. Four compounds (anabasine, anatabine, nornicotine, and scopoletin) were identified and quantified in 3 N. glauca flowers extracts, establishing them as potential sources of alien bio-molecules. The most abundant constituent was anabasine, determined at 3900 µg/g in the methanolic extract. These extracts were utilized as feeding treatments on Apis mellifera honeybees, resulting in mild toxicity documented by 16-18% mortality. A slightly increased effect was elicited by the methanolic extract containing anabasine at 20 µg/mL, where mortality approached 25%. Dead bees were screened for residues of the N. glauca flower extracts compounds and a significant mean concentration of anabasine was evidenced in both 10 and 20 µg/mL treatments, ranging from 51 to 92 ng/g per bee body weight. Scopoletin was also detected in trace amounts. CONCLUSIONS: The mild toxicity of the extracts in conjunction with the alkaloid and coumarin residual detection in bees, suggest that these alien bio-molecules are transferred within the food chain, suggesting a chemical invasion phenomenon, never reported before.

Adverse Effects in Skeletal Muscle Following the Medicinal Use of Nicotiana glauca.

Nicotiana glauca is a cosmopolitan shrub, used in medicine to treat swellings, wounds, sores and cancer. However, its users lack of knowledge of the adverse effects. We seek to evaluate the effects of lipid extracts from N. glauca on myoblasts, identifying the compounds which cause undesirable effects. Myoblasts are important in muscle homeostasis, thus a high death rate of them cause myopathies. We performed an ethanolic extraction from leaves of N. glauca and the extract was successively partitioned with hexane, chloroform and ethyl acetate. The effects of extracts in C2C12 cells were analysed by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL), Mitotracker and 4',6-diamidino-2-phenylindole (DAPI) staining, Western blotting, real-time PCR and immunofluorescence assays. Caspase activity was studied. The fraction with the highest apoptotic effects was analysed by chromatography, NMR and GC-MS spectrometry were used to identify the apoptotic agent, after which its biological activity was evaluated. The extracts from N. glauca induced apoptosis in C2C12 cells involving caspase-3/7. We found that the extracts trigger a defence response in muscle through Akt and heat shock protein 27 (HSP27). We identified an apoptotic agent as palmitic acid. These data suggest that the use of N. glauca in hormone replacement therapy, or in other therapies affects skeletal muscle homeostasis, worsening the negative effects of the menopause. Thus, the relevance of this work lies in the fact that it is the first time that a report about the molecular mechanism responsible for the side effects of medicinal use of N. glauca, has been shown. Moreover the compound responsible for these effects has been identified.

Analysis of Fitness Trade-Offs in the Host Range Expansion of an RNA Virus, Tobacco Mild Green Mosaic Virus.

The acquisition of new hosts provides a virus with more opportunities for transmission and survival but may be limited by across-host fitness trade-offs. Major causes of across-host trade-offs are antagonistic pleiotropy, that is, host differential phenotypic effects of mutations, a Genotype x Environment interaction, and epistasis, a Genotype x Genotype interaction. Here, we analyze if there are trade-offs, and what are the causes, associated with the acquisition by tobacco mild green mosaic virus (TMGMV) of a new host. For this, the multiplication of sympatric field isolates of TMGMV from its wild reservoir host Nicotiana glauca and from pepper crops was quantified in the original and the heterologous hosts. TMGMV isolates from N. glauca were adapted to their host, but pepper isolates were not adapted to pepper, and the acquisition of this new host was associated with a fitness penalty in the original host. Analyses of the collection of field isolates and of mutant genotypes derived from biologically active cDNA clones showed a role of mutations in the coat protein and the 3' untranslated region in determining within-host virus fitness. Fitness depended on host-specific effects of these mutations, on the genetic background in which they occurred, and on higher-order interactions of the type Genotype x Genotype x Environment. These types of effects had been reported to generate across-host fitness trade-offs under experimental evolution. Our results show they may also operate in heterogeneous natural environments and could explain why pepper isolates were not adapted to pepper and their lower fitness in N. glaucaIMPORTANCE The acquisition of new hosts conditions virus epidemiology and emergence; hence it is important to understand the mechanisms behind host range expansion. Experimental evolution studies have identified antagonistic pleiotropy and epistasis as genetic mechanisms that limit host range expansion, but studies from virus field populations are few. Here, we compare the performance of isolates of tobacco mild green mosaic virus from its reservoir host, Nicotiana glauca, and its new host, pepper, showing that acquisition of a new host was not followed by adaptation to it but was associated with a fitness loss in the original host. Analysis of mutations determining host-specific virus multiplication identified antagonistic pleiotropy, epistasis, and host-specific epistasis as mechanisms generating across-host fitness trade-offs that may prevent adaptation to pepper and cause a loss of fitness in N. glauca Thus, mechanisms determining trade-offs, identified under experimental evolution, could also operate in the heterogeneous environment in which natural plant virus populations occur.

Respiratory Failure Due To Plant Poisoning: Nicotiana glauca Graham.

BACKGROUND: Cases of human poisoning associated with Nicotiana glauca Graham are quite rare. Patients may consume the plant by mistaking it for spinach or use it as a folk remedy. The toxin stimulates the acetylcholine receptors in the central and autonomic nervous systems. A prompt and appropriate supportive treatment is crucial for survival. CASE REPORT: A 72-year-old female with a known breast cancer was brought to our emergency department (ED) with complaints of nausea and vomiting. The patient stated that her complaints started 2 h after she had cooked and eaten a plant in the hopes that it would help cure her cancer. On the second hour after arrival, the patient became confused. It was observed that the chest wall movements had decreased and that the patient had difficulty breathing. Due to respiratory failure, orotracheal intubation was performed to secure the airway. A sample of the plant that was eaten was requested from the patient's relatives. According to an Internet search, the plant sample was identified as N. glauca Graham. The patient was diagnosed with N. glauca Graham poisoning. The patient was extubated after 30 h. On 3-month follow-up, the patient was neurologically intact and had no complaints related to poisoning. WHY SHOULD AN EMERGENCY PHYSICIAN BE AWARE OF THIS?: Patients with incurable diseases such as cancer tend to try folk remedies and can end up in ED. In patients who present with muscle paralysis and respiratory failure, emergency physicians should consider N. glauca Graham as a cause of plant poisoning.

Nicotiana glauca whole-genome investigation for cT-DNA study.

OBJECTIVE: Nicotiana glauca (tree tobacco) is a naturally transgenic plant, containing sequences acquired from Agrobacterium rhizogenes by horizontal gene transfer. Besides, N. glauca contains a wide profile of alkaloids of medical interest. DATA DESCRIPTION: We report a high-depth sequencing and de novo assembly of N. glauca full genome and analysis of genome elements with bacterial origin. The draft genome assembly is 3.2 Gb, with N50 size of 31.1 kbp. Comparative analysis confirmed the presence of single, previously described gT insertion. No evidence was acquired to support idea of multiple T-DNA insertions in the N. glauca genome. Our data is the first comprehensive de novo assembly of tree tobacco and provide valuable information for researches in pharmacological and in phylogenetic fields.

Horizontal Transfer of a Synthetic Metabolic Pathway between Plant Species.

Transgene expression from the plastid (chloroplast) genome provides unique advantages, including high levels of foreign protein accumulation, convenient transgene stacking in operons, and increased biosafety due to exclusion of plastids from pollen transmission [1, 2]. However, applications in biotechnology and synthetic biology are severely restricted by the very small number of plant species whose plastid genomes currently can be transformed [3, 4]. Here we report a simple method for the introduction of useful plastid transgenes into non-transformable species. The transgenes tested comprised a synthetic operon encoding three components of a biosynthetic pathway for producing the high-value ketocarotenoid astaxanthin in the plastids of the cigarette tobacco, Nicotiana tabacum. Transplastomic N. tabacum plants accumulated astaxanthin to up to 1% of the plants' dry weight. We then used grafting, a procedure recently shown to facilitate horizontal genome transfer between plants [5-7], to let the transgenic chloroplast genome move across the graft junction from N. tabacum plants into plants of the nicotine-free tree species Nicotiana glauca. Transplastomic N. glauca trees expressing the synthetic pathway were recovered at high frequency, thus providing a straightforward method for extension of the transplastomic technology to new species.

From Microhabitat of Floral Nectar Up to Biogeographic Scale: Novel Insights on Neutral and Niche Bacterial Assemblies.

Microbial model systems are very useful in addressing macro-ecological questions. Two major theories exist to date, to explain the community structure of organisms: (1) the dispersal (neutral) assembly theory which predicts that community similarity decreases with increasing geographic distance, independent of any environmental variables, and (2) the niche assembly theory which predicts that the communities' compositions are more homogeneous among sites characterized by similar environmental conditions. Our study system offered a unique opportunity to investigate the relative role of environmental conditions and spatial factors in shaping community composition. We explored the bacterial community composition (BCC) of Nicotiana glauca floral nectar using the Illumina MiSeq technique at three spatial scales (plants, site, and region) and two taxonomic levels. Floral nectar samples were collected from 69 N. glauca plants at 11 different sites along a 200-km transect in Israel, along three biogeographic regions. A distance decay of BCC was found among all plants throughout Israel, but such pattern was not found among either sites or biogeographical regions. The BCC was also governed by environmental conditions in all examined scales (from the plant up to the biogeographical region). We also found that taxonomic resolution (89 and 97% sequence identity for clustering operational taxonomic units) affected the results of these BCC analyses. Hence, our study revealed that the BCC in N. glauca floral nectar is shaped by both the environmental conditions and the distance between plants, depending on the sampling scale under examination as well as by taxonomic resolution.

Ethnobotanical inventory of medicinal plants used in the Qampaya District, Bolivia / Inventario etnobotánico de plantas medicinales usadas en el Distrito Qampaya, Bolivia

Herbal medicine therapy is traditionally practiced by indigenous healers in Bolivia for hundreds years. Due to the great geographical and ecological diversity, there are thousands of native plants, which are utilized against diverse types of diseases. Nowadays, there is a worldwide problem connected with possible loss of ethnobotany knowledge because of the lack of the interest of young people. In present study, survey focused on medicinal plants used by rural people in Qampaya District, Potosí Department, Bolivia has been done. The data were collected from 60 respondents by semi-structured interviews. The results showed that 60 plant species belonging to 30 families are known as curative plants in this area. The mostly named families were Asteraceae (14 species) followed by Lamiaceae (7 species) and Brassicaceae (4 species). Predominant health problems treated by these plants are urological problems and gastro-intestinal disorders. The most frequently used plant parts were leaves and the preparation is mostly done as infusion. Even though the knowledge of using medical plants plays important role in life of Bolivian rural people, which use plants as medicines against various types of diseases, this study showed that 25 percent of respondents didn’t know any medicinal plants. On the other hand 40 percent of asked people have known 6 or more medicinal plant species.

La terapia de la medicina herbolaria es practicada, tradicionalmente, por los curanderos indígenas en Bolivia, desde hace cientos de años. Gracias a la gran diversidad geográfica y ecológica, hay miles de plantas nativas, que se utilizan para el tratamiento de diferentes tipos de enfermedades. Hoy en día, hay un problema mundial relacionado con la posible pérdida del conocimiento etnobotánico, esto debido a la falta de interés de las nuevas generaciones. El presente estudio descriptivo, mediante una encuesta, fue enfocado a las plantas medicinales utilizadas por la población rural del Distrito Qampaya, Departamento de Potosí, Bolivia. Los datos se obtuvieron de 60 informantes mediante una encuesta semiestructurada. Los resultados mostraron que 60 especies de plantas medicinales. pertenecientes a 30 familias botánicas, son conocidas en el área de intervención. Las familias botánicas, más importantes de uso medicinal son Asteraceae (14 especies), seguido de Lamiaceae (7 especies) y Brassicaceae (4 especies). Los problemas predominantes de salud, tratados con estas especies, son los urológicos y trastornos gastrointestinales. La parte de la planta más utilizada son las hojas y la forma más común de uso es en infusión. A pesar de que el conocimiento sobre la utilización de las plantas medicinales, juega un papel importante en la vida de la población rural de Bolivia, quienes utilizan las plantas medicinales para el tratamiento de diversos tipos de enfermedades, este estudio mostró que el 25 por ciento de los encuestados no tienen conocimiento sobre la utilidad medicinal de ninguna especie vegetal. Por otro lado, el 40 por ciento de los informantes conocen 6 o más especies de plantas medicinales.

Scopoletin, an active principle of tree tobacco (Nicotiana glauca) inhibits human tumor vascularization in xenograft models and modulates ERK1, VEGF-A, and FGF-2 in computer model.

We recently reported the antineovascularization effect of scopoletin on rat aorta and identified its potential anti-angiogenic activity. Scopoletin could be useful as a systemic chemotherapeutic agent against angiogenesis-dependent malignancies if its antitumorigenic activity is investigated and scientifically proven using a suitable human tumor xenograft model. In the present study, bioassay-guided (anti-angiogenesis) phytochemical investigation was conducted on Nicotiana glauca extract which led to the isolation of scopoletin. Further, anti-angiogenic activity of scopoletin was characterized using ex vivo, in vivo and in silico angiogenesis models. Finally, the antitumorigenic efficacy of scopoletin was studied in human colorectal tumor xenograft model using athymic nude mice. For the first time, an in vivo anticancer activity of scopoletin was reported and characterized using xenograft models. Scopoletin caused significant suppression of sprouting of microvessels in rat aortic explants with IC50 (median inhibitory concentration) 0.06µM. Scopoletin (100 and 200mg/kg) strongly inhibited (59.72 and 89.4%, respectively) vascularization in matrigel plugs implanted in nude mice. In the tumor xenograft model, scopoletin showed remarkable inhibition on tumor growth (34.2 and 94.7% at 100 and 200mg/kg, respectively). Tumor histology revealed drastic reduction of the extent of vascularization. Further, immunostaining of CD31 and NG2 receptors in the histological sections confirmed the antivascular effect of scopoletin in tumor vasculature. In computer modeling, scopoletin showed strong ligand affinity and binding energies toward the following angiogenic factors: protein kinase (ERK1), vascular endothelial growth factor A (VEGF-A), and fibroblast growth factor 2 (FGF-2). These results suggest that the antitumor activity of scopoletin may be due to its strong anti-angiogenic effect, which may be mediated by its effective inhibition of ERK1, VEGF-A, and FGF-2.