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1.
Biotechnol Lett ; 44(2): 333-340, 2022 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-35182262

RESUMO

OBJECTIVE: Unlike plant cell suspension culture, the proliferation of callus in bioreactors has received inadequate attention. The magnificent potential of plant callus becomes more appreciated as the research unfolds and promises interesting applications including the production of valuable metabolites, therapeutic antibodies, bioactive extracts with regenerating effects, and the generation of genetically improved plants. Issues such as the lack of 3D-access of the cells to the nutrients, using an interfering gelling substance as the support matrix, and the changes in the medium formulation during the growth phase were discouraging factors for extending research on this topic. Considering the existing drawbacks, a novel open-flow spray bioreactor (OFSB) was configured to circumvent the associated problems with the solid cell culture and promote the applicability of plant callus culture via improving the feeding strategy. METHODS: Applying similar subculture conditions, the proliferation of Arnebia pulchra and Hyoscyamus niger calli as the examples of two important plant families (Boraginaceae and Solanaceae) was studied in the OFSB in comparison with similar calli that grew in Petri dishes and jars. RESULTS: A. pulchra and H. niger calli obtained the weight gains of (%87.3 and %106.7) in the Petri dishes, (%208.7 and %226) in the jars, and (%288.6 and %320.0) in OFSB, respectively, while no significant changes were observed in the productivity indices of the examined calli. CONCLUSION: The simple design of OFSB bypasses most of the notorious problems associated with solid plant callus culture. OFSB technical features allow the bioreactor to be used for growth optimization of various types of plant calli in a cost-effective manner.


Assuntos
Boraginaceae , Hyoscyamus , Reatores Biológicos , Proliferação de Células , Meios de Cultura , Humanos
2.
Biomed Res Int ; 2021: 9940591, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34381841

RESUMO

This study is aimed at identifying the chemical composition of the essential oil extracted from the Syzygium aromaticum seeds, as well as investigating its biological activities, insecticide effect, and allelopathic properties. The extraction yield was about 14.3 and 7.14% for grounded and ungrounded seeds, respectively. The GC-MS analysis allowed the identification of 17 heterogeneous compounds, including eugenol (68.7-87.4%), as major compound, cyperene (20.5-7.2%), phenethyl isovalerate (6.4-3.6%), and cis-thujopsene (1.9-0.8%), respectively, for grounded and ungrounded seeds. Concerning the antibacterial activity, the diameter of the inhibition zone reached 35 mm when the essential oil extracted from grounded seeds was applied against Escherichia coli. Regarding the antioxidant activity via the DPPH radical scavenging test, the IC50 varied from 1.2 ± 0.1 to 2.8 ± 0.5 µg/mL. With respect to reducing power, the efficient concentration EC50 ranged from 32 to 50 µg/mL. The essential oil exhibited also an allelopathic effect against seeds of Hyoscyamus niger, as well as an insecticide effect against Sitophilus oryzae with a DL50 value of 252.4 µL/L air. These findings enhance the use of this spice as a natural food preservative and encourage its use in several fields, including pharmaceutical, cosmetics, agriculture, and therapy, that could be a strategic way to guarantee the consumer's health.


Assuntos
Antibacterianos/química , Antioxidantes/química , Inseticidas/química , Óleos Voláteis/química , Syzygium/química , Animais , Antibacterianos/isolamento & purificação , Antibacterianos/farmacologia , Antioxidantes/isolamento & purificação , Antioxidantes/farmacologia , Testes de Sensibilidade a Antimicrobianos por Disco-Difusão , Escherichia coli/efeitos dos fármacos , Escherichia coli/crescimento & desenvolvimento , Inocuidade dos Alimentos , Cromatografia Gasosa-Espectrometria de Massas , Hyoscyamus/efeitos dos fármacos , Hyoscyamus/crescimento & desenvolvimento , Inseticidas/isolamento & purificação , Inseticidas/farmacologia , Viabilidade Microbiana/efeitos dos fármacos , Óleos Voláteis/isolamento & purificação , Óleos Voláteis/farmacologia , Óleos Vegetais/química , Óleos Vegetais/isolamento & purificação , Sementes/química , Gorgulhos/efeitos dos fármacos , Gorgulhos/crescimento & desenvolvimento
3.
Cell Commun Signal ; 19(1): 61, 2021 05 25.
Artigo em Inglês | MEDLINE | ID: mdl-34034759

RESUMO

BACKGROUND: Chronic superphysiological glucose and insulin concentrations are known to trigger several tissue and organ failures, including insulin resistance, oxidative stress and chronic low-grade inflammation. Hence, the screening for molecules that may counteract such conditions is essential in current existing therapeutic strategies, thereby the use of medicinal plant derivatives represents a promising axis in this regard. METHODS: In this study, the effect of a selected traditional medicinal plant, Hyoscyamus albus from which, calystegines have been isolated, was investigated in an experimental model of hyperinsulinemia and hyperglycemia induced on HepG2 cells. The mRNA and protein expression levels of different insulin signaling, gluconeogenic and inflammatory pathway- related molecules were examined. Additionally, cell viability and apoptosis, oxidative stress extent and mitochondrial dysfunctions were assayed using flow cytometric and qRT-PCR techniques. RESULTS: Treatment of IR HepG2 cells with calystegines strongly protected the injured cells from apoptosis, oxidative stress and mitochondrial integrity loss. Interestingly, nortropane alkaloids efficiently regulated the impaired glucose metabolism in IR HepG2 cells, through the stimulation of glucose uptake and the modulation of SIRT1/Foxo1/G6PC/mTOR pathway, which is governing the hepatic gluconeogenesis. Furthermore, the alkaloidal extract restored the defective insulin signaling pathway, mainly by promoting the expression of Insr at the mRNA and protein levels. What is more, treated cells exhibited significant mitigated inflammatory response, as evidenced by the modulation and the regulation of the NF- κB/JNK/TLR4 axis and the downstream proinflammatory cytokines recruitment. CONCLUSION: Overall, the present investigation demonstrates that calystegines from Hyoscyamus albus provide cytoprotection to the HepG2 cells against insulin/glucose induced insulin resistance and apoptosis due to the regulation of SIRT1/Foxo1/G6PC/mTOR and NF-κB/JNK/TLR4 signaling pathways. Video Abstract.


Assuntos
Hyoscyamus/química , Hiperglicemia/tratamento farmacológico , Hiperinsulinismo/tratamento farmacológico , Sistema de Sinalização das MAP Quinases , NF-kappa B/metabolismo , Nortropanos/uso terapêutico , Sirtuína 1/metabolismo , Apoptose/efeitos dos fármacos , Caspases/metabolismo , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Citocinas/metabolismo , Gluconeogênese/efeitos dos fármacos , Glucose/metabolismo , Células Hep G2 , Humanos , Inflamação/metabolismo , Inflamação/patologia , Mediadores da Inflamação/metabolismo , Insulina/metabolismo , Resistência à Insulina , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/patologia , Nortropanos/farmacologia , Estresse Oxidativo , Extratos Vegetais/farmacologia , Extratos Vegetais/uso terapêutico , Sementes/química , Transdução de Sinais/efeitos dos fármacos , Serina-Treonina Quinases TOR/metabolismo
4.
Phytother Res ; 35(7): 4000-4006, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-33860587

RESUMO

The outbreak of Coronavirus disease 2019 (COVID-19) has caused a global health crisis. Nevertheless, no antiviral treatment has yet been proven effective for treating COVID-19 and symptomatic supportive cares have been the most common treatment. Therefore, the present study was designed to evaluate the effects of propolis and Hyoscyamus niger L. extract in patients with COVID-19. This randomized clinical trial was conducted on 50 cases referred to Akhavan and Sepehri Clinics, Kashan university of medical sciences, Iran. Subjects were divided into two groups (intervention and placebo). This syrup (containing 1.6 mg of methanolic extract along with 450 mg of propolis per 10 mL) was administered three times a day to each patient for 6 days. The clinical symptoms of COVID-19 such as: dry cough, shortness of breath, sore throat, chest pain, fever, dizziness, headache, abdominal pain, and diarrhea were reduced with propolis plus Hyoscyamus niger L. extract than the placebo group. However, the administration of syrup was not effective in the control of nausea and vomiting. In conclusion, syrup containing propolis and Hyoscyamus niger L. extract had beneficial effects in ameliorating the signs and symptoms of COVID-19 disease, in comparison with placebo groups.


Assuntos
COVID-19 , Hyoscyamus , Extratos Vegetais/uso terapêutico , Própole , Síndrome do Desconforto Respiratório/tratamento farmacológico , Adulto , COVID-19/tratamento farmacológico , Feminino , Humanos , Hyoscyamus/química , Irã (Geográfico) , Masculino , Metanol , Pessoa de Meia-Idade , Própole/uso terapêutico , Síndrome do Desconforto Respiratório/virologia , Resultado do Tratamento
5.
Pak J Biol Sci ; 23(10): 1260-1266, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-32981259

RESUMO

BACKGROUND AND OBJECTIVE: The use of herbicides leads to severe damage, especially to the environment. The aim of this investigation was operated to study the allelopathic effect of Calotropis procera, Hyoscyamus muticus and Pulicaria undulata aqueous extracts on the seed germination of Portulaca oleracea and Chenopodium murale. MATERIALS AND METHODS: The aerial parts of the plants (Calotropis procera, Hyoscyamus muticus and Pulicaria undulata) washed well in distilled water and were dried in room temperature then ground to fine powder. Aqueous extract was prepared using distilled water. RESULTS: In this study, different concentrations of plant extracts (1, 3 and 5%) were used to illustrate which of these concentrations have the potential to inhibit seed growth of Portulaca oleracea and Chenopodium murale. Concentration of 5% for all plant extracts used to inhibited the growth of all seeds of Chenopodium murale while, concentration of 5% for Pulicaria undulata extract only inhibited the growth of all seeds of Portulaca oleracea. CONCLUSION: This work dealt with the use of allelopathy strategy in preventing the growth of some weeds to be one of the safest alternative ways of using herbicides. The results showed the effective effect of plant extracts for both Calotropis procera, Hyoscyamus muticus and Pulicaria undulata on the seed germination of Portulaca oleracea and Chenopodium murale.


Assuntos
Calotropis/metabolismo , Chenopodium/efeitos dos fármacos , Germinação/efeitos dos fármacos , Hyoscyamus/metabolismo , Extratos Vegetais/farmacologia , Portulaca/efeitos dos fármacos , Pulicaria/metabolismo , Sementes/efeitos dos fármacos , Alelopatia , Herbicidas , Plantas Daninhas/efeitos dos fármacos , Temperatura
6.
Plant Physiol Biochem ; 155: 416-428, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32814278

RESUMO

Species of Hyoscyamus are rich sources of medicinally important tropane alkaloids, which have anticholinergic, antispasmodic and sedative effects and are competitive inhibitors of acetylcholine. The application of nanotechnology and nanomaterials for elicitation is rapidly expanding and recent research indicates that silicon dioxide nanoparticles (SiO2 NPs) can be used as an efficient elicitor to increase the production of hyoscyamine and scopolamine in Hyoscyamus species. Thus, in this work, the effect of SiO2 NPs (0, 25, 50, 100 and 200 mg L-1) with two treatment times (24 and 48 h) on the growth rate, total phenol and flavonoid content (TPC, TFC), antioxidant enzyme activity, tropane alkaloid yield and pmt (putrescine N-methyltransferase) and h6h (hyoscyamine 6-hydroxylase) gene expression levels in hairy roots of two Hyoscyamus species (H. reticulatus and H. pusillus) was investigated. The highest TPC and TFC accumulation was obtained in H. reticulatus elicited by SiO2 NPs (100 and 200 mg L-1), respectively, at 24 h of treatment. High-performance liquid chromatography (HPLC) revealed the highest amount of hyoscyamine (140.15 µg g-1 FW) and scopolamine (67.71 µg g-1 FW) accumulated in H. reticulatus transformed roots treated with 100 mg L-1 SiO2 NPs at 24 h, with a respective increase of 1212% and 272% compared to non-treated roots. In H. pusillus, the highest hyoscyamine (7.42 µg g-1 FW) and scopolamine (15.56 µg g-1 FW) production (about 82% and 241% higher, respectively, compared to the lowest amounts) was achieved with 25 and 100 mg L-1 SiO2 NPs, respectively, at 48 h of treatment. Semi-quantitative RT-PCR analysis determined the highest expression level of pmt and h6h genes in H. reticulatus transformed roots supplemented with 100 mg L-1 SiO2 NPs.


Assuntos
Hyoscyamus/metabolismo , Nanopartículas , Raízes de Plantas/metabolismo , Dióxido de Silício/farmacologia , Tropanos/metabolismo , Regulação da Expressão Gênica de Plantas , Hyoscyamus/genética , Raízes de Plantas/genética , Técnicas de Cultura de Tecidos
7.
PLoS One ; 15(5): e0231355, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32437389

RESUMO

The overexploitation of medicinal plants is depleting gene pool at an alarming rate. In this scenario inducing the genetic variability through targeted mutations could be beneficial in generating varieties with increased content of active compounds. The present study aimed to develop a reproducible protocol for in vitro multiplication and mutagenesis of Hyoscyamus niger targeting putrescine N-methyltransferase (PMT) and 6ß-hydroxy hyoscyamine (H6H) genes of alkaloid biosynthetic pathway. In vitro raised callus were treated with different concentrations (0.01% - 0.1%) of Ethyl Methane Sulfonate (EMS). Emerging multiple shoots and roots were obtained on the MS media supplemented with cytokinins and auxins. Significant effects on morphological characteristics were observed following exposure to different concentrations of EMS. EMS at a concentration of 0.03% was seen to be effective in enhancing the average shoot and root number from 14.5±0.30 to 22.2 ±0.77 and 7.2±0.12 to 8.8±0.72, respectively. The lethal dose (LD50) dose was calculated at 0.08% EMS. The results depicted that EMS has an intense effect on PMT and H6H gene expression and metabolite accumulation. The transcripts of PMT and H6H were significantly upregulated at 0.03-0.05% EMS compared to control. EMS treated explants showed increased accumulation of scopolamine (0.639 µg/g) and hyoscyamine (0.0344µg/g) compared to untreated.


Assuntos
Metanossulfonato de Etila/toxicidade , Hiosciamina/metabolismo , Hyoscyamus/crescimento & desenvolvimento , Metiltransferases/genética , Oxigenases de Função Mista/genética , Mutagênese , Mutação , Escopolamina/metabolismo , Vias Biossintéticas , Regulação da Expressão Gênica de Plantas , Hyoscyamus/efeitos dos fármacos , Hyoscyamus/genética , Hyoscyamus/metabolismo , Mutagênicos/toxicidade , Plantas Geneticamente Modificadas/efeitos dos fármacos , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/crescimento & desenvolvimento , Plantas Geneticamente Modificadas/metabolismo
8.
Toxicon ; 177: 52-88, 2020 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-32217234

RESUMO

Datura stramonium, Atropa belladonna, Hyoscyamus niger, and Scopolia carniolica are all temperate plants from the family Solanaceae, which as a result of their anticholinergic tropane alkaloids, hyoscyamine/atropine and scopolamine, have caused many cases of poisoning around the world. Despite the danger these nightshade plants represent, the literature often presents incomplete cases lacking in details and filled with ambiguity, and reviews on the topic tend to be limited in scope. Many also point to a gap in knowledge of these plants among physicians. To address this, the following review focuses on intoxications involving these plants as reported in the literature between 1966 and 2018, with brief mention to pertinent related plants to contextualise and provide a fuller picture of the situation surrounding the presently discussed temperate plants. Analysis of the literature displays that D. stramonium is largely associated with drug use among teens while A. belladonna is primarily ingested as a result of the berries being mistaken for edible fruits. H. niger was found to be largely ingested when mistaken for other plants, and S. carniolica was the cause of incredibly few intoxications.


Assuntos
Antagonistas Colinérgicos/toxicidade , Extratos Vegetais/toxicidade , Intoxicação por Plantas/epidemiologia , Solanaceae , Alcaloides de Solanáceas/toxicidade , Alcaloides , Atropa belladonna , Europa (Continente) , Humanos , Hyoscyamus , Escopolamina , Scopolia , Tropanos
10.
J Ethnopharmacol ; 244: 112151, 2019 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-31404578

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: The Norse berserkers were wild warriors of Scandinavia known to enter a trance-like state that allowed them to fight with increased strength and a rage that granted them immunity to many forms of harm in battle. Though many theories have been advanced as to the cause of this state, the most widely believed is that the intoxicating mushroom Amanita muscaria was used. AIM OF THE STUDY: The following article underlines the issues with this theory and provides an alternate intoxicant that fits with the reports of berserker behaviour much better: Hyoscyamus niger. MATERIALS AND METHODS: Literature from a variety of disciplines pertaining to history, toxicology, pharmacology, and botany was compiled to frame and support the argument. RESULTS: H. niger proved to be a more likely intoxicant used to induce the berserker rage state. CONCLUSIONS: With its anticholinergic tropane alkaloids and symptom profile, H. niger is a much more likely cause of the berserker state than A muscaria. Though there is not enough archaeological and historical evidence to prove or disprove this theory, it provides a novel explanation that is at present the most viable means of understanding the berserkers' trance.


Assuntos
Hyoscyamus , Amanita , Etnobotânica , Humanos , Preparações de Plantas/toxicidade , Países Escandinavos e Nórdicos
11.
Pak J Pharm Sci ; 32(2): 661-667, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31081780

RESUMO

The work presented in this paper illustrates the isolation and structure elucidation of secondary metabolites of Hyoscyamus albus. Two new natural source and three known compounds were isolated from the Hyoscyamus albus. Among the isolated compounds, grivilloside H (1) and betulaplatoside (2) were isolated for the first time while scopolamine (3), ß-sitosterol (4) and stigmasterol (5) have been reported previously from the same plant. The structures of all the isolated compounds were established by using modern spectroscopic technique (UV, IR, NMR, and EI-MS) and by comparing with those available in literature.


Assuntos
Hyoscyamus/metabolismo , Compostos Fitoquímicos/química , Plantas Medicinais/metabolismo , Glucosídeos/química , Glucosídeos/isolamento & purificação , Glucosídeos/metabolismo , Hyoscyamus/química , Espectroscopia de Ressonância Magnética , Estrutura Molecular , Compostos Fitoquímicos/isolamento & purificação , Compostos Fitoquímicos/metabolismo , Plantas Medicinais/química , Escopolamina/química , Escopolamina/isolamento & purificação , Escopolamina/metabolismo , Metabolismo Secundário , Sitosteroides/química , Sitosteroides/isolamento & purificação , Sitosteroides/metabolismo , Espectrofotometria Infravermelho , Espectrofotometria Ultravioleta , Estigmasterol/química , Estigmasterol/isolamento & purificação , Estigmasterol/metabolismo
12.
Plant Mol Biol ; 100(4-5): 433-450, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30968307

RESUMO

KEY MESSAGE: Cybrid plant mitochondria undergo homologous recombination, mainly BIR, keep a single allele for each gene, and maintain exclusive sequences of each parent and a single copy of the homologous regions. The maintenance of a dynamic equilibrium between the mitochondrial and nuclear genomes requires continuous communication and a high level of compatibility between them, so that alterations in one genetic compartment need adjustments in the other. The co-evolution of nuclear and mitochondrial genomes has been poorly studied, even though the consequences and effects of this interaction are highly relevant for human health, as well as for crop improvement programs and for genetic engineering. The mitochondria of plants represent an excellent system to understand the mechanisms of genomic rearrangements, chimeric gene formation, incompatibility between nucleus and cytoplasm, and horizontal gene transfer. We carried out detailed analyses of the mtDNA of a repeated cybrid between the solanaceae Nicotiana tabacum and Hyoscyamus niger. The mtDNA of the cybrid was intermediate between the size of the parental mtDNAs and the sum of them. Noticeably, most of the homologous sequences inherited from both parents were lost. In contrast, the majority of the sequences exclusive of a single parent were maintained. The mitochondrial gene content included a majority of N. tabacum derived genes, but also chimeric, two-parent derived, and H. niger-derived genes in a tobacco nuclear background. Any of these alterations in the gene content could be the cause of CMS in the cybrid. The parental mtDNAs interacted through 28 homologous recombination events and a single case of illegitimate recombination. Three main homologous recombination mechanisms were recognized in the cybrid mitochondria. Break induced replication (BIR) pathway was the most frequent. We propose that BIR could be one of the mechanisms responsible for the loss of the majority of the repeated regions derived from H. niger.


Assuntos
Genoma Mitocondrial , Hibridização Genética , Mitocôndrias/genética , DNA Mitocondrial/química , Genoma de Planta , Recombinação Homóloga , Hyoscyamus/genética , Tabaco/genética
13.
Acta Biol Hung ; 69(4): 437-448, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30587015

RESUMO

Hyoscyamus reticulatus L. is a herbaceous biennial belonging to the solanaceae family. Hyoscyamine and scopolamine as main tropane alkaloids accumulated in henbane are widely used in medicine to treat diseases such as parkinson's or to calm schizoid patients. Hairy roots media manipulation which uses elicitors to activate defense mechanisms is one of the main strategies for inducing secondary metabolism as well as increasing the production of valuable metabolites. Cotyledon-derived hairy root cultures were transformed by Agrobacterium rhizogenes. Sodium nitroprusside (SNP), a nitric oxide donor), was used in various concentrations (0, 50, 100, 200 and 300 µM) and exposure times (24 and 48 h). Treatment with SNP led to a significant reduction in fresh and dry weight of hairy roots, compared to control cultures. ANOVA results showed that elicitation of hairy root cultures with SNP at different concentrations and exposure times significantly affected the activity of as antioxidant enzymes such as catalase (CAT), peroxidase (POD) and ascorbate peroxidase (APX). The highest hyoscyamine and scopolamine production (about 1.2-fold and 1.5-fold increases over the control) was observed at 50 and 100 µM SNP at 48 and 24 hours of exposure time, respectively. This is the first report of SNP elicitation effects on the production of tropane alkaloids in hairy root cultures.


Assuntos
Antioxidantes/metabolismo , Enzimas/biossíntese , Hyoscyamus/efeitos dos fármacos , Doadores de Óxido Nítrico/farmacologia , Nitroprussiato/farmacologia , Proteínas de Plantas/biossíntese , Raízes de Plantas/efeitos dos fármacos , Tropanos/metabolismo , Agrobacterium/genética , Agrobacterium/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Relação Dose-Resposta a Droga , Indução Enzimática , Hyoscyamus/enzimologia , Hyoscyamus/crescimento & desenvolvimento , Hyoscyamus/microbiologia , Raízes de Plantas/enzimologia , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/microbiologia , Fatores de Tempo , Técnicas de Cultura de Tecidos , beta-Glucosidase/genética , beta-Glucosidase/metabolismo
14.
Sci Rep ; 8(1): 17969, 2018 12 19.
Artigo em Inglês | MEDLINE | ID: mdl-30568179

RESUMO

Under hypoxic conditions, the expression of Vitreoscilla hemoglobin (VHb) in plants is proposed to increase the productivity of certain oxygen-requiring metabolic pathways by promoting the delivery of oxygen. Tropane alkaloids (TAs) are a class of important plant secondary metabolites with significant medicinal value; the final step in their biosynthesis requires oxygen. Whether heterologous expression of VHb, especially in different subcellular compartments, can accelerate the accumulation of TAs is not known. Herein, the effect of heterologous expression of VHb in different subcellular locations on the TA profile of H. niger hairy roots was investigated. The targeted expression of VHb in the plastids (using pVHb-RecA construct), led to the accumulation of 197.68 µg/g hyoscyamine in the transgenic H. niger hairy roots, which was 1.25-fold of the content present in the lines in which VHb expression was not targeted, and 3.66-fold of that present in the wild type (WT) lines. The content of scopolamine was increased by 2.20- and 4.70-fold in the pVHb-RecA transgenic lines compared to that in the VHb transgenic and WT lines. Our results demonstrate that VHb could stimulate the accumulation of TAs in the transgenic H. niger hairy roots. Quantitative RT-PCR analysis revealed that the expression of key genes involved in TA biosynthesis increased significantly in the VHb transgenic lines. We present the first description of a highly efficient strategy to increase TA content in H. niger. Moreover, our results also shed light on how the production of desired metabolites can be efficiently enhanced by using more accurate and appropriate genetic engineering strategies.


Assuntos
Proteínas de Bactérias/genética , Expressão Gênica , Hyoscyamus/fisiologia , Raízes de Plantas/fisiologia , Tropanos/metabolismo , Hemoglobinas Truncadas/genética , Proteínas de Bactérias/metabolismo , Vias Biossintéticas , Regulação da Expressão Gênica de Plantas , Espaço Intracelular , Modelos Biológicos , Plantas Geneticamente Modificadas/genética , Transporte Proteico , Transformação Genética , Tropanos/química , Hemoglobinas Truncadas/metabolismo
15.
Plant Physiol Biochem ; 127: 47-54, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29549757

RESUMO

N-methylputrescine is the precursor of nicotine and pharmaceutical tropane alkaloids such as hyoscyamine. Putrescine N-methyltransferase (PMT) catalyzes the N-methylation of putrescine to form N-methylputrescine. While the role of PMT in nicotine biosynthesis is clear, knowledge of PMT in the biosynthesis of tropane alkaloids (TAs) and the regulation of polyamines remains limited. We characterized a PMT gene from Hyoscyamus niger, designated HnPMT that was specifically expressed in roots, especially in the secondary roots and dramatically induced by methyl jasmonate (MeJA). The GUS gene was specifically expressed in Arabidopsis roots or in the vascular tissues, including pericycles and endodermis, of the H. niger hairy root cultures, when it was driven by the 5'-flanking promoter region of HnPMT. The recombinant HnPMT was purified for enzymatic assays. HnPMT converted putrescine to form N-methylputrescine, as confirmed by LC-MS. The kinetics analysis revealed that HnPMT had high affinity with putrescine but low catalytic activity, suggesting that it was a rate-limiting enzyme. When HnPMT was suppressed in the H. niger plants by using the VIGS approach, the contents of N-methylputrescine and hyoscyamine were markedly decreased, but the contents of putrescine, spermidine and a mixture of spermine and thermospermine were significantly increased; this suggested that HnPMT was involved in the biosynthesis of tropane alkaloids and played a competent role in regulating the biosynthesis of polyamines. Functional identification of HnPMT facilitated the understanding of TA biosynthesis and thus implied that the HnPMT-catalyzed step might be a target for metabolic engineering of the TA production in H. niger.


Assuntos
Hyoscyamus , Metiltransferases , Raízes de Plantas , Arabidopsis/enzimologia , Arabidopsis/genética , Hyoscyamus/enzimologia , Hyoscyamus/genética , Metiltransferases/genética , Metiltransferases/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Raízes de Plantas/genética , Raízes de Plantas/metabolismo , Plantas Geneticamente Modificadas/enzimologia , Plantas Geneticamente Modificadas/genética
16.
Plant Cell Rep ; 36(10): 1615-1626, 2017 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-28707113

RESUMO

KEY MESSAGE: Tetraploidy improves overexpression of h6h and scopolamine production of H. muticus, while in H. senecionis, pmt overexpression and elicitation can be used as effective methods for increasing tropane alkaloids. The effects of metabolic engineering in a polyploid context were studied by overexpression of h6h in the tetraploid hairy root cultures of H. muticus. Flow cytometry analysis indicated genetic stability in the majority of the clones, while only a few clones showed genetic instability. Among all the diploid and tetraploid clones, the highest level of h6h transgene expression and scopolamine accumulation was interestingly observed in the tetraploid clones of H. muticus. Therefore, metabolic engineering of the tropane biosynthetic pathway in polyploids is suggested as a potential system for increasing the production of tropane alkaloids. Transgenic hairy root cultures of Hyoscyamus senecionis were also established. While overexpression of pmt in H. senecionis was correlated with a sharp increase in hyoscyamine production, the h6h-overexpressing clones were not able to accumulate higher levels of scopolamine than the leaves of intact plants. Applying methyl jasmonate was followed by a sharp increase in the expression of pmt and a drop in the expression of tropinone reductase II (trII) which consequently resulted in the higher biosynthesis of hyoscyamine and total alkaloids in H. senecionis.


Assuntos
Alcaloides/metabolismo , Hyoscyamus/genética , Engenharia Metabólica/métodos , Raízes de Plantas/genética , Ploidias , Tropanos/metabolismo , Vias Biossintéticas/genética , Diploide , Regulação da Expressão Gênica de Plantas , Genes de Plantas/genética , Hyoscyamus/classificação , Hyoscyamus/metabolismo , Raízes de Plantas/metabolismo , Plantas Geneticamente Modificadas , Escopolamina/metabolismo , Especificidade da Espécie , Tetraploidia , Técnicas de Cultura de Tecidos
17.
Yao Xue Xue Bao ; 52(1): 172-9, 2017 Jan.
Artigo em Chinês | MEDLINE | ID: mdl-29911833

RESUMO

Tropane alkaloids are anticholinergic drugs widely used clinically. Biosynthesis of tropane alkaloids in planta involves a step of transamination of phenylalanine. Based on the sequenced transcriptomes of lateral roots and leaves of Hyoscyamus niger, we found three annotated aromatic amino acid aminotransferases, which were respectively named HnArAT1, HnArAT2 and HnArAT3. Sequence analysis showed that HnArAT3 had highest similarity with the reported Atropa belladonna Ab Ar AT4, which was involved in tropane alkaloid(TA) to provide the precursor of the phenyllactic acid moiety. Tissue expression pattern analysis indicated that HnArAT3 was specifically expressed in lateral roots, where is the organ synthesizing tropane alkaloids. Then, method of virus induced gene silencing (VIGS) was used to characterize the function of HnArAT3 in H. niger. Gene expression analysis given by real-time quantitative PCR showed that all the transgenic lines had lower expression levels of HnArAT3 than the non-transgenic control, and HPLC analysis of alkaloids demonstrated significant decrease in the contents of hyoscyamine, anisodamine and scopolamine in planta. These results suggested that HnArAT3 was involved in the phenyllactic acid branch of TA biosynthetic pathway. Molecular cloning and functional identification of HnArAT3 laid the foundation for further understanding of TA biosynthesis and metabolic regulation, and also provided a new candidate gene for engineering biosynthetic pathway of tropane alkaloids.


Assuntos
Alcaloides/biossíntese , Hyoscyamus/genética , Proteínas de Plantas/genética , Transaminases/genética , Tropanos/metabolismo , Atropa belladonna , Vias Biossintéticas , Antagonistas Colinérgicos , Clonagem Molecular , Hiosciamina , Hyoscyamus/enzimologia , Raízes de Plantas/enzimologia , Raízes de Plantas/genética , Escopolamina , Alcaloides de Solanáceas
18.
J Hazard Mater ; 324(Pt B): 306-320, 2017 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-27810325

RESUMO

In this study, seeds of Hyoscyamus niger were exposed to different concentrations (50-800µgmL-1) of single-walled carbon nanotubes (SWCNTs) under different levels of drought stress (0.5-1.5MPa) for 14days. Germinated seeds were subsequently allowed to grow in the same culture media for 7 more days to test the further response of the seedlings in terms of biochemical changes to the employed treatments. Seeds subjected to drought showed reduction in germination percentage, vigor and lengths of roots and shoots. However, inclusion of SWCNTs at the two lowest concentrations significantly alleviated the drought stress (up to moderate levels only)-induced reduction in germination and growth attributes. This happened due to increased water uptake, up-regulation of mechanisms involved in starch hydrolysis, and reduction in oxidative injury indices including H2O2, malondialdehyde contents and electrolyte leakage. The improved plant performance under PEG-induced drought stress was a consequence of changes in the expression of various antioxidant enzymes including SOD, POD, CAT, and APX, and also biosynthesis of proteins, phenolics, and specific metabolites such as proline. Results demonstrate that treatment by low concentrations of SWCNTs can induce tolerance in seedlings against low to moderate levels of drought through enhancing water uptake and activating plant defense system.


Assuntos
Secas , Hyoscyamus/efeitos dos fármacos , Nanotubos de Carbono , Estresse Fisiológico/efeitos dos fármacos , Catalase/metabolismo , Germinação/efeitos dos fármacos , Peróxido de Hidrogênio/metabolismo , Hyoscyamus/crescimento & desenvolvimento , Hyoscyamus/metabolismo , Malondialdeído/metabolismo , Peroxidases/metabolismo , Fenóis/metabolismo , Proteínas de Plantas/metabolismo , Brotos de Planta/efeitos dos fármacos , Brotos de Planta/crescimento & desenvolvimento , Brotos de Planta/metabolismo , Polietilenoglicóis/toxicidade , Sementes/efeitos dos fármacos , Sementes/crescimento & desenvolvimento , Superóxido Dismutase/metabolismo
19.
São Paulo; s.n; 2017. 35 p. tab.
Tese em Espanhol | HomeoIndex - Homeopatia | ID: hom-11706

RESUMO

Relato de um paciente portador de transtorno afetivo bipolar, em uso de medicação psicotrópica e em psicoterapia regular, que, após iniciar uso de Hyoscyamus niger 30 CH, apresentou melhora importante da sintomatologia maníaca, demonstrada através de redução de pontuação da Escala de Avaliação de mania de Bech-Rafaelsen. Portanto, o medicamento homeopático é uma opção de tratamento para o transtorno bipolar do humor e outros transtornos mentais graves.(AU)


A report of a patient with bipolar affective disorder, using psychotropic medication and regular psychotherapy, who, after starting Hyoscyamus niger 30 CH, presented an important improvement of the manic symptomatology, demonstrated through reduction of the Mania Rating Scale Of Bech-Rafaelsen. Therefore, the homeopathic medicine is a treatment option for bipolar disorder of mood and other serious mental disorders. (AU)


Assuntos
Humanos , Masculino , Pessoa de Meia-Idade , Transtorno Bipolar , Homeopatia , Hyoscyamus/uso terapêutico , Terapêutica
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