RESUMO
The plant hormone jasmonic acid (JA) fine tunes the growth-defense dilemma by inhibiting plant growth and stimulating the accumulation of secondary compounds. We investigated the interactions between JA and phytochrome B signaling on growth and the accumulation of selected secondary metabolites in Hypericum perforatum L., a medically important plant, by spraying plants with methyl jasmonate (MeJA) and by adding far-red (FR) lighting. MeJA inhibited plant growth, decreased fructose concentration, and enhanced the accumulation of most secondary metabolites. FR enhanced plant growth and starch accumulation and did not decrease the accumulation of most secondary metabolites. MeJA and FR acted mostly independently with no observable interactions on plant growth or secondary metabolite levels. The accumulation of different compounds (e.g., hypericin, flavonols, flavan-3-ols, and phenolic acid) in shoots, roots, and root exudates showed different responses to the two treatments. These findings indicate that the relationship between growth and secondary compound accumulation is specific and depends on the classes of compounds and/or their organ location. The combined application of MeJA and FR enhanced the accumulation of most secondary compounds without compromising plant growth. Thus, the negative correlations between biomass and the content of secondary compounds predicted by the growth-defense dilemma were overcome.
Assuntos
Ciclopentanos/farmacologia , Hypericum/crescimento & desenvolvimento , Hypericum/metabolismo , Luz , Oxilipinas/farmacologia , Exsudatos de Plantas/metabolismo , Folhas de Planta/metabolismo , Raízes de Plantas/metabolismo , Acetatos/farmacologia , Biomassa , Vias Biossintéticas/efeitos dos fármacos , Carboidratos/análise , Hypericum/efeitos dos fármacos , Hypericum/efeitos da radiação , Íons , Tamanho do Órgão/efeitos dos fármacos , Fenóis/análise , Pigmentos Biológicos/metabolismo , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/efeitos da radiação , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/efeitos da radiaçãoRESUMO
We report a new green route for preparing MnO2/perlite nanocomposites (NCs) by leaf extract of Hypericum perforatum. Characterization of the physicochemical properties of the MnO2/perlite-NCs was performed using XRD, FESEM, EDX, FT-IR, and DLS techniques. Furthermore, their effects on the phytochemical classification and growth parameters of H. perforatum shoot cultures were assessed. According to the FESEM image, the synthesized spherical MnO2 nanoparticles on the sheet-like structure of nano-perlite were formed, ranging about 20-50 nm. In addition, based on the EDX spectra, the elemental analysis showed the presence of Carbon, Oxygen, Silicon, Aluminum, and Manganese elements in the as-synthesized MnO2/perlite-NCs. Biological studies confirmed that nano-perlite and MnO2/perlite-NCs were non-toxic to H. perforatum shoot cultures and showed positive effects on plant growth in specific concentrations. Overall, phytochemical classification demonstrated that the terpenoids decreased in the evaluated treatments, while hypericin and pseudohypericin were increased in some treatments (25, 50 and 150 mg/L of nano-perlite) relative to control. Metabolomics results suggested that both nano-perlite and MnO2/perlite-NCs can be used as elicitors and new nanofertilizers for generating some secondary metabolites.
Assuntos
Hypericum/fisiologia , Manganês/metabolismo , Nanocompostos , Óxido de Alumínio , Antracenos , Hypericum/crescimento & desenvolvimento , Compostos de Manganês , Nanopartículas , Óxidos/farmacologia , Perileno/análogos & derivados , Compostos Fitoquímicos/farmacologia , Óleos de Plantas , Dióxido de Silício , Espectroscopia de Infravermelho com Transformada de FourierRESUMO
Hypericum perforatum L. commonly known as Saint John's Wort (SJW), is an important medicinal plant that has been used for more than 2000 years. Although H. perforatum produces several bioactive compounds, its importance is mainly linked to two molecules highly relevant for the pharmaceutical industry: the prenylated phloroglucinol hyperforin and the naphtodianthrone hypericin. The first functions as a natural antidepressant while the second is regarded as a powerful anticancer drug and as a useful compound for the treatment of Alzheimer's disease. While the antidepressant activity of SJW extracts motivate a multi-billion dollar industry around the world, the scientific interest centers around the biosynthetic pathways of hyperforin and hypericin and their medical applications. Here, we focus on what is known about these processes and evaluate the possibilities of combining state of the art omics, genome editing, and synthetic biology to unlock applications that would be of great value for the pharmaceutical and medical industries.
Assuntos
Hypericum/química , Hypericum/genética , Compostos Fitoquímicos/biossíntese , Compostos Fitoquímicos/farmacologia , Extratos Vegetais/farmacologia , Proteínas de Plantas/genética , Antracenos , Antidepressivos/farmacologia , Antineoplásicos/farmacologia , Europa (Continente) , Humanos , Hypericum/crescimento & desenvolvimento , Hypericum/metabolismo , Perileno/análogos & derivados , Perileno/farmacologia , Floroglucinol/análogos & derivados , Floroglucinol/farmacologia , Terpenos/farmacologiaRESUMO
The R2R3-MYB family is one of the largest families of plant transcription factor playing significant roles in plant growth. Although this gene family has been studied in many species, the R2R3-MYBs in Hypericum perforatum which is the first sequenced species in Malpighiales have not been analyzed. A total of 109 R2R3-MYB genes were identified in H. perforatum and clustered into 36 clades. Gene Ontology analysis revealed that most of the R2R3-MYB genes were involved in biological processes. Four kinds of cis-acting elements were found within the promoter regions, the majority of which were related to the stress responses and plant growth/development. The transcriptome data of different tissues (root, stems, leaves, and flowers) showed that the spatial expression profiles of R2R3-MYBs were different. Also, real-time quantitative PCR analysis revealed that eleven stress-related R2R3-MYB genes showed specific expression patterns under diverse treatments. In addition, sub-cellular localization analysis indicated that five significant proteins HpMYB45, HpMYB48, HpMYB55, HpMYB63, and HpMYB70 were all localized in the nucleus. This study was the first report on identification and characterization of R2R3-MYB gene family in H. perforatum. It facilitated the identification of tissue-preferential and stress-related genes and provided deep insights into the function of R2R3-MYBs in H. perforatum.
Assuntos
Regulação da Expressão Gênica de Plantas , Genoma de Planta , Hypericum/genética , Proteínas de Plantas/genética , Estresse Fisiológico/genética , Fatores de Transcrição/genética , Temperatura Baixa , Secas , Flores/efeitos dos fármacos , Flores/genética , Flores/crescimento & desenvolvimento , Regulação da Expressão Gênica no Desenvolvimento , Ontologia Genética , Hypericum/efeitos dos fármacos , Hypericum/crescimento & desenvolvimento , Anotação de Sequência Molecular , Família Multigênica , Filogenia , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/genética , Folhas de Planta/crescimento & desenvolvimento , Proteínas de Plantas/classificação , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/genética , Raízes de Plantas/crescimento & desenvolvimento , Caules de Planta/efeitos dos fármacos , Caules de Planta/genética , Caules de Planta/crescimento & desenvolvimento , Regiões Promotoras Genéticas , Isoformas de Proteínas/genética , Salinidade , Cloreto de Sódio/farmacologia , Fatores de Transcrição/classificaçãoRESUMO
In the 60 years since Skoog and Miller first reported the chemical redirection of plant growth the underlying biochemical mechanisms are still poorly understood, with one challenge being the capacity for applied growth regulators to act indirectly or be metabolized to active phytohormones. We hypothesized that tryptophan is metabolized to auxin, melatonin or serotonin inducing organogenesis in St. John's wort (Hypericum perforatum L.). Root explants from two germplasm lines of St. John's wort with altered melatonin metabolism and wildtype were incubated with auxin or tryptophan for 24, 48 or 72 h to induce regeneration. In wildtype, tryptophan had little effect on the indoleamine pathway, and was found to promote primary growth, suggesting excess tryptophan moved quickly through various secondary metabolite pathways and protein synthesis. In lines 4 and 112 tryptophan was associated with modified morphogenesis, indoleamine and auxin levels. Incubation with tryptophan increased shoot organogenesis while incubation with auxin led to root regeneration. The established paradigm of thought views tryptophan primarily as a precursor for auxin and indoleamines, among other metabolites, and mediation of auxin action by the indoleamines as a one-way interaction. We propose that these processes run in both directions with auxin modifying indoleamine biosynthesis and the melatonin:serotonin balance contributing to its effects on plant morphogenesis, and that tryptophan also functions as an inductive signal to mediate diverse phytochemical and morphogenetic pathways.
Assuntos
Aminas/metabolismo , Hypericum/efeitos dos fármacos , Ácidos Indolacéticos/farmacologia , Triptofano/metabolismo , Hypericum/crescimento & desenvolvimento , Hypericum/metabolismo , Ácidos Indolacéticos/metabolismo , Indóis/química , Modelos Lineares , Melatonina/metabolismo , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/metabolismo , Brotos de Planta/efeitos dos fármacos , Brotos de Planta/crescimento & desenvolvimento , Brotos de Planta/metabolismo , Serotonina/metabolismo , Triptofano/farmacologiaRESUMO
It appears that the biologically-synthesized nanoparticles (NPs) have potential to perform as effective elicitors for the production of valuable secondary metabolites in plants. Besides, it has been reported that the toxicity of the biologically-synthesized NP is not as much as that of the chemically-synthesized NPs. Therefore, it is necessary to test their advantages aspects. In this study, the physical synthesis of perlite NPs and biologically-synthesis of TiO2/perlite nanocomposites (NCs) were conducted. Subsequently, their effects and explant source influence on the growth characteristics and secondary metabolite profiles of Hypericum perforatum callus cultures were evaluated. According to the obtained results, morphology of the synthesized perlite NPs and TiO2/perlite NCs were mesoporous and spherical with sizes ranging about 14.51-23.34 and 15.50-24.61 nm, respectively. Addition of perlite NPs and TiO2/perlite NCs to the culture medium at the concentration range of 25-200 mg/L showed no adverse impacts on the growth characteristics of H. perforatum calli. According to the GC-MS analysis, the stress caused by perlite NPs and TiO2/perlite NCs led to an increase in the variety, amount and number of volatile compounds. The calli obtained from in vitro grown plants produced more volatile compounds relative to the calli obtained from field grown plants under the nanomaterial stress conditions. The production of hypericin and pseudohypericin were also determined in the callus cultures under desired nanomaterials elicitation. Accordingly, our results suggest that perlite NPs and TiO2/perlite NCs can possibly be considered as effective elicitors for the production of volatile compounds, hypericin, and pseudohypericin in callus cultures of H. perforatum.
Assuntos
Óxido de Alumínio/química , Hypericum/metabolismo , Nanopartículas/química , Compostos Fitoquímicos/metabolismo , Caules de Planta/crescimento & desenvolvimento , Dióxido de Silício/química , Titânio/química , Compostos Orgânicos Voláteis/metabolismo , Antracenos , Hypericum/crescimento & desenvolvimento , Perileno/análogos & derivados , Perileno/metabolismoRESUMO
Microbiomes can dramatically alter individual plant performance, yet how these effects influence higher-order processes is not well resolved. In particular, little is known about how microbiome effects on individual plants alter plant population dynamics, a question critical to imperiled species conservation. Here we integrate bioassays, multidecadal demographic data, and integral projection modeling to determine how the presence of the natural soil microbiome underlies plant population dynamics. Simulations indicated that the presence of soil microbiomes boosted population growth rates (λ) of the endangered Hypericum cumulicola by 13% on average, the difference between population growth versus decline in 76% of patches. The greatest benefit (47% increase in λ) occurred in low-nutrient, high-elevation habitats, suggesting that the soil microbiome may help expand H. cumulicola's distribution to include these stressful habitats. Our results demonstrate that soil microbiomes can significantly affect plant population growth and persistence and support the incorporation of soil microbiomes into conservation planning.
Assuntos
Hypericum/crescimento & desenvolvimento , Microbiota , Microbiologia do Solo , Altitude , Espécies em Perigo de Extinção , Germinação/fisiologia , Hypericum/fisiologia , Modelos EstatísticosRESUMO
Naphthodianthrone derivatives that produced in Hypericum perforatum (St. John's Wort) are valuable secondary metabolites for depression treatment and photodynamic therapy. However, the traditional cultivation of this plant does not meet both quantitatively and qualitatively the high demand of the pharmaceutical industry. So, the adventitious root culture along with elicitation has been introduced as an alternative for production of such valuable bioactive compounds. The aim of this study was to evaluate the effect of darkness and red, blue and fluorescent light on growth and production of secondary metabolites in the adventitious root cultivation of H. perforatum. Our results showed that biomass production was significantly higher in the cultures grown under dark and red light, but in terms of hypericins production, red light was the best. Despite the inhibitory effect of five weeks blue light treatment on both biomass and secondary metabolite production of adventitious roots, one-week blue light treatment of four-weeks grown roots is an effective stimulator for increasing total phenolic compounds and hypericins. Interestingly, the roots were regenerated under red light and stems and leaves were formed.
Assuntos
Hypericum/crescimento & desenvolvimento , Hypericum/metabolismo , Luz , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/efeitos da radiação , Metabolismo Secundário/efeitos da radiação , Antracenos , Biomassa , Meios de Cultura , Flavonoides/metabolismo , Hypericum/efeitos da radiação , Perileno/análogos & derivados , Perileno/metabolismo , Fenóis/metabolismo , Açúcares/metabolismoRESUMO
The purpose of this study was to assess the antioxidant and anti-inflammatory potential of liposomal formulations enriched with Hypericum hookerianum (Hyp) aqueous extracts. Cotyledon segments derived from protocorms of H. hookerianum were cultured on Murashige and Skoog (MS) media supplemented with Kinetin (KN, 1 mgl-1) and Naphthalene acetic acid (NAA, 0.1 mgl-1) to induce hypericin-rich red shoots (HypR, 0.87 mg/G DW). Highly stable liposomes (-29.4 mV) were successfully developed which encapsulated 63 ± 0.8% Hyp extracts, respectively. MTT assay subsequently confirmed the biocompatibility of liposome compositions using fibroblast cell lines. This work also evaluated acute toxicity of L-HypR and L-HypG formulations using Danio rerio (Zebrafish) embryos for 96 hpf. The expression of antioxidant and anti-inflammatory genes were found to be upregulated for L-HypR than L-HypG (green shoots without hypericin) formulations. These properties of L-HypR may be extremely useful for incorporating lipophilic substances into the food or pharmaceutical industry.
Assuntos
Anti-Inflamatórios/farmacologia , Antioxidantes/farmacologia , Hypericum/química , Perileno/análogos & derivados , Extratos Vegetais/farmacologia , Animais , Antracenos , Anti-Inflamatórios/administração & dosagem , Anti-Inflamatórios/química , Antioxidantes/administração & dosagem , Antioxidantes/química , Linhagem Celular , Humanos , Hypericum/crescimento & desenvolvimento , Lipossomos/química , Perileno/administração & dosagem , Perileno/química , Perileno/farmacologia , Extratos Vegetais/administração & dosagem , Extratos Vegetais/química , Peixe-Zebra/embriologiaRESUMO
Hypericum perforatum is a perennial herb that produces the anti-depression metabolite hypericin (Hyp). While several efforts to increase Hyp production have been made, the effects of temperatures on growth and Hyp biosynthesis are still limited. In this study, the growth morphophysiological traits, Hyp biosynthesis and their related genes expression, as well as major bioactive compounds accumulation and antioxidant capacity were assessed by exposing H. perforatum seedlings to three different temperatures (15, 22 and 30⯰C). The results showed that aerial parts biomass was greater at 15⯰C with 1.3 and 1.6-fold increase compared to at 22 and 30⯰C, in large part because of greater increase in chlorophyll content, stem number and leaf area on a per plant basis. Hyp content in the aerial parts was greater 1.9 and 5.6-fold on a per plant basis compared to 22 and 30⯰C treatments, and the contents of other bioactive compounds (flavonoids and phenolics) as well as antioxidant capacity in the aerial parts, on dry weight and per plant basis, also exhibited significant increases with the temperatures decrease. The mRNA expressions of eight genes (psbA, psbB, psbC, psbD, ycf3, ycf4, ycf5 and matK) related to photosynthesis and two genes (Polyketide synthase, PKS; Phenolic oxidative coupling protein, Hyp-1) involved in Hyp biosynthesis were also up-regulated at 15⯰C. The findings are useful in guiding cultivation and regulating Hyp biosynthesis in H. perforatum.
Assuntos
Hypericum/crescimento & desenvolvimento , Hypericum/metabolismo , Perileno/análogos & derivados , Antracenos , Hypericum/fisiologia , Perileno/metabolismo , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/metabolismo , Folhas de Planta/fisiologia , Proteínas de Plantas/metabolismo , TemperaturaRESUMO
Melatonin and serotonin are important signaling and stress mitigating molecules that play important roles across growth and development in plants. Despite many well-documented responses, a systematic investigation of the entire metabolic pathway (tryptophan, tryptamine, and N-acetylserotonin) does not exist, leaving many open questions. The objective of this study was to determine the responses of Hypericum perforatum (L.) to melatonin, serotonin, and their metabolic precursors. Two well-characterized germplasm lines (#4 and 112) created by mutation and a haploid breeding program were compared to wild type to identify specific responses. Germplasm line 4 has lower regenerative and photosynthetic capacity than either wild type or line 112, and there are documented significant differences in the chemistry and physiology of lines 4 and 112. Supplementation of the culture media with tryptophan, tryptamine, N-acetylserotonin, serotonin, or melatonin partially reversed the regenerative recalcitrance and growth impairment of the germplasm lines. Quantification of phytohormones revealed crosstalk between the indoleamines and related phytohormones including cytokinin, salicylic acid, and abscisic acid. We hypothesize that melatonin and serotonin function in coordination with their metabolites in a cascade of phytochemical responses including multiple pathways and phytohormone networks to direct morphogenesis and protect photosynthesis in H. perforatum.
Assuntos
Hypericum/crescimento & desenvolvimento , Hypericum/metabolismo , Melatonina/metabolismo , Desenvolvimento Vegetal/fisiologia , Serotonina/metabolismo , Hypericum/efeitos dos fármacos , Melatonina/farmacologia , Desenvolvimento Vegetal/efeitos dos fármacos , Reguladores de Crescimento de Plantas/metabolismo , Reguladores de Crescimento de Plantas/farmacologia , Sementes/crescimento & desenvolvimento , Serotonina/farmacologia , Transdução de Sinais/fisiologiaRESUMO
Hypericin, an important determinant of the pharmacological properties of the genus Hypericum, is considered as a major molecule for drug development. However, biosynthesis and accumulation of hypericin is not well understood. Identification of genes differentially expressed in tissues with and without hypericin accumulation is a useful strategy to elucidate the mechanisms underlying the development of the dark glands and hypericin biosynthesis. Suppression Subtractive Hybridization (SSH) is a unique method for PCR-based amplification of specific cDNA fragments that differ between a control (driver) and experimental (tester) transcriptome. This technique relies on the removal of dsDNA formed by hybridization between a control and test sample, thus eliminating cDNAs of similar abundance, and retaining differentially expressed or variable in sequence cDNAs. In our laboratory we applied this method to identify the genes involved in the development of dark glands and accumulation of hypericin in Hypericum perforatum. Here we describe the complete procedure for the construction of hypericin gland-specific subtracted cDNA library.
Assuntos
Biblioteca Gênica , Hypericum/genética , Hypericum/metabolismo , Perileno/análogos & derivados , Técnicas de Hibridização Subtrativa/métodos , Antracenos , Vias Biossintéticas , DNA Complementar/genética , Germinação , Hypericum/anatomia & histologia , Hypericum/crescimento & desenvolvimento , Perileno/análise , Perileno/metabolismo , Reação em Cadeia da Polimerase/métodos , RNA Mensageiro/genética , RNA de Plantas/genéticaRESUMO
Context The genus Hypericum (Hypericaceae) has attracted remarkable scientific interest as its members have yielded many bioactive compounds. Objective The current study presents investigations on the accumulation of hypericin, pseudohypericin, hyperforin, adhyperforin, chlorogenic acid, neochlorogenic acid, caffeic acid, 2,4-dihydroxybenzoic acid, 13,118-biapigenin, hyperoside, isoquercitrin, quercitrin, quercetin, avicularin, rutin, (+)-catechin and (-)-epicatechin in seven Hypericum (Hypericaceae) species growing wild in Turkey, namely, H. aviculariifolium Jaup. and Spach subsp. aviculariifolium (Freyn and Bornm.) Robson var. albiflorum (endemic), H. bithynicum Boiss., H. calycinum L., H. cardiophyllum Boiss., H. elongatum L. subsp. microcalycinum (Boiss. and Heldr.) Robson, H. hirsutum L. and H. xylosteifolium (Spach) N. Robson. Materials and methods The plant materials were collected at flowering period and dissected in different tissues. Air-dried plant material including stems, leaves and flowers was mechanically powdered with a laboratory mill and samples (0.1 g) were extracted in 10 mL of 100% methanol by ultrasonication at 40 °C for 30 min for HPLC-PDA analyses. Results Accumulation levels of the investigated compounds varied greatly depending on species and plant part. Discussion For the first time, the detailed chemical profiles of corresponding Turkish Hypericum species were reported and the results were discussed from a phytochemical point of view. Conclusions The present data have importance in evaluation of plant resources of Hypericum genus in selecting the new potential sources of bioactive compounds.
Assuntos
Hypericum/metabolismo , Extratos Vegetais/metabolismo , Fracionamento Químico/métodos , Cromatografia Líquida de Alta Pressão , Hypericum/classificação , Hypericum/crescimento & desenvolvimento , Fitoterapia , Componentes Aéreos da Planta , Plantas Medicinais , Turquia , UltrassomRESUMO
BACKGROUND: Cancer is an unnatural type of tissue growth in which the cells exhibit unrestrained division, leading to a progressive increase in the number of dividing cells. It is now the second largest cause of death in the world. The present study concerned antioxidant, anticancer and anticholinesterase activities and protocatechuic, catechin, caffeic acid, syringic acid, p-coumaric acid and o-coumaric concentrations in methanol extracts of flowers, fruits and seeds of Hypericum amblysepalum. MATERIALS AND METHODS: Antioxidant properties including free radical scavenging activity and reducing power, and amounts of total phenolic compounds were evaluated using different tests. Protocatechuic, catechin, caffeic acid, syringic acid, p-coumaric acid and o-coumaric concentrations in extracts were determined by HPLC. Cytotoxic effects were determined using the MTT test with human cervix cancer (HeLa) and rat kidney epithelium cell (NRK-52E) lines. Acetyl and butyrylcholinesterase inhibitory activities were measured by by Ellman method. RESULTS: Total phenolic content of H. amblysepalum seeds was found to be higher than in fruit and flower extracts. DPPH free radical scavenging activity of the obtained extracts gave satisfactory results versus butylated hydroxyanisole and butylated hydroxytoluene as controls. Reducing power activity was linearly proportional to the studied concentration range: 10-500 µg/ mL LC50 values for H. amblysepalum seeds were 11.7 and 2.86 respectively for HeLa and NRK-52E cell lines. Butyryl-cholinesterase inhibitory activity was 76.9±0.41 for seed extract and higher than with other extracts. CONCLUSIONS: The present results suggested that H. amblysepalum could be a potential candidate anti-cancer drug for the treatment of human cervical cancer, and good source of natural antioxidants.
Assuntos
Antineoplásicos/farmacologia , Antioxidantes/farmacologia , Inibidores da Colinesterase/farmacologia , Flores/química , Frutas/química , Hypericum/crescimento & desenvolvimento , Extratos Vegetais/farmacologia , Sementes/química , Acetilcolinesterase/química , Animais , Compostos de Bifenilo/química , Butirilcolinesterase/química , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Cromatografia Líquida de Alta Pressão , Sequestradores de Radicais Livres/farmacologia , Células HeLa , Humanos , Hypericum/metabolismo , Neoplasias/tratamento farmacológico , Neoplasias/patologia , Picratos/química , RatosRESUMO
Physiology, oxidative stress and production of metabolites in Hypericum perforatum exposed to moderate Cd and/or La concentration (10 µM) were studied. La evoked increase in reactive oxygen species, malondialdehyde and proline but suppressed growth, tissue water content, glutathione, ascorbic acid and affected mineral nutrient contents more than Cd while the impact of Cd+La was not synergistic. Similar trend was observed at the level of superoxide dismutase gene expression. Shoot Cd amount increased in Cd+La while only root La increased in the same treatment. Extensive quantification of secondary metabolites revealed that La affected phenolic acids more pronouncedly than Cd in shoots and roots. Flavonols were suppressed by La that could contribute to the appearance of oxidative damage. Procyanidins increased in response to La in the shoots but decreased in the roots. Metabolic responses in Cd+La treatment resembled those of La treatment (almost identically in the roots). Phenylalanine ammonia-lyase activity was mainly suppressed by La. The presence of La also depleted amount of hypericin and expression of its putative gene (hyp-1) showed similar trend but accumulation of hyperforin increased under Cd or La excess. Clear differences in the stem and root anatomy in response to Cd or La were also found. Overall, H. perforatum is La-sensitive species and rather Cd ameliorated negative impact of La.
Assuntos
Cádmio/toxicidade , Poluentes Ambientais/toxicidade , Hypericum/metabolismo , Lantânio/toxicidade , Estresse Oxidativo/efeitos dos fármacos , Biomassa , Flavonoides/metabolismo , Hidroxibenzoatos/metabolismo , Hypericum/efeitos dos fármacos , Hypericum/crescimento & desenvolvimento , Microscopia de Fluorescência , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/metabolismo , Plântula/efeitos dos fármacos , Plântula/crescimento & desenvolvimento , Plântula/metabolismoRESUMO
Hypericum perforatum L. (St. John's wort), a perennial flowering plant native to Europe, is widely used as a medicinal plant and has a long history of its use in the treatment of various ailments. Currently, H. perforatum is widely used as an herbal remedy for the treatment of mild to moderate depression. Hypericins are natural napthodianthrone compounds produced from H. perforatum (St. John's wort) which are having antitumor, antiviral (i.e., against human immunodeficiency and hepatitis C virus), antineoplastic, and antidepressant properties. Currently, field-grown plant materials are generally used for the commercial production of hypericins. It has been reported that hypericin accumulation in natural plants is influenced by different ecological and environmental conditions including light intensity, nitrogen availability, temperature, seasons, and growing regions. Therefore, up to 17-fold and 13-fold differences in hypericin and pseudohypericin amounts, respectively, are reported in different phytopharmaceutical preparations. Plant cell and organ cultures are effective systems for producing natural products, and attempts were made for the production of biomass and stable concentrations of hypericins through in vitro cultures of H. perforatum. Cell, callus, shoot, plantlet, and adventitious root cultures have been established and various chemical and physical factors which influence the biomass and secondary metabolite accumulation have been investigated. Large-scale plantlet and adventitious root cultures have also been attempted in H. perforatum in bioreactors, and various strategies have been applied for the production of higher biomass and secondary products. This review describes the biotechnological approaches employed for the production of hypericins and focuses upon the challenges and future prospects.
Assuntos
Produtos Biológicos/metabolismo , Biotecnologia/métodos , Hypericum/crescimento & desenvolvimento , Hypericum/metabolismo , Perileno/análogos & derivados , Tecnologia Farmacêutica/métodos , Antracenos , Técnicas de Cultura de Células , Perileno/metabolismoRESUMO
Hypericum perforatum L. (St. John's Wort) is an important medicinal plant which is widely used in the treatment for depression and irritable bowel syndrome. It is also used as a dietary supplement. Major bioactive phytochemicals of H. perforatum are phenolics and flavonoids. Quality of these phytochemicals is dramatically influenced by environmental and biological factors in the field grown plants. As an alternative, we have developed adventitious root cultures in large-scale bioreactors for the production of useful phytochemicals. Adventitious roots of H. perforatum were cultured in 500 l pilot-scale airlift bioreactors using half-strength Murashige and Skoog medium with an ammonium and nitrate ratio of 5:25 mM and supplemented with 1.0 mg l(-1) indole butyric acid, 0.1 mg l(-1) kinetin, and 3 % sucrose for the production of bioactive phenolics and flavonoids. Then 4.6 and 6.3 kg dry biomass were realized in the 500 l each of drum-type and balloon-type bioreactors, respectively. Accumulation of 66.9 mg g(-1) DW of total phenolics, 48.6 mg g(-1) DW of total flavonoids, 1.3 mg g(-1) DW of chlorogenic acid, 0.01 mg g(-1) DW of hyperin, 0.04 mg g(-1) DW of hypericin, and 0.01 mg g(-1) DW of quercetin could be achieved with adventitious roots cultured in 500 l balloon-type airlift bioreactors. Our findings demonstrate the possibilities of using H. perforatum adventitious root cultures for the production of useful phytochemicals to meet the demand of pharmaceutical and food industry.
Assuntos
Reatores Biológicos , Hypericum/crescimento & desenvolvimento , Biomassa , Cromatografia Líquida de Alta Pressão , Flavonoides/análise , Hypericum/química , Hypericum/metabolismo , Fenóis/análise , Projetos PilotoRESUMO
Species of the genus Hypericum contain a rich array of unusual polyketides, however, only a small proportion of the over 450 Hypericum species, other than the popular medicinal supplement St. John's Wort (Hypericum perforatum), have even been chemically characterized. Hypericum gentianoides, a small annual used medicinally by Cherokee Americans, contains bioactive acylphloroglucinols. Here, we identify acylphloroglucinol constituents of H. gentianoides and determine a potential pathway to their synthesis. Liquid chromatography/electrospray ionization-mass spectrometry (LC/ESI-MS) and HPLC-UV indicate that the level of accumulation and profile of acylphloroglucinols in H. gentianoides vary little seasonally when grown in a greenhouse, but do vary with development and are highly dependent on the accession, highlighting the importance of the selection of plant material for study. We identify the chemical structures of the nine prevalent polyketides, based on LC/ESI-MS and hybrid quadrupole orthogonal time-of-flight (Q-TOF) mass spectrometry; these metabolites include one monomeric phlorisobutyrophenone (PIB) derivative and eight dimeric acylphloroglucinols. Q-TOF spectrometry was used to identify eight additional PIB derivatives that were not detected by LC/ESI-MS. These data lead us to propose that diacylphloroglucinols are synthesized via modification of PIB to yield diverse phloroglucinol and filicinic acids moieties, followed by dimerization of a phloroglucinol and a filicinic acid monomer to yield the observed complement of diacylphloroglucinols. The metabolomics data from H. gentianoides are accessible in plant metabolomics resource (PMR) (http://www.metnetdb.org/pmr), a public metabolomics database with analysis software for plants and microbial organisms.
Assuntos
Hypericum/metabolismo , Floroglucinol/metabolismo , Vias Biossintéticas , Cromatografia Líquida , Ecótipo , Hypericum/crescimento & desenvolvimento , Íons , Floroglucinol/química , Floroglucinol/isolamento & purificação , Extratos Vegetais/metabolismo , Espectrometria de Massas por Ionização por ElectrosprayRESUMO
OBJECTIVES: This study was carried out to assess the anti-obesity effect of Hypericum silenoidesâ Juss. and Hypericum philonotisâ Cham. & Schlecht. in male Wistar rats fed with a cafeteria diet. METHODS: Adult male Wistar rats with an initial body weight of 290-320 g were used in this trial. The rats were fed with a cafeteria diet for 77 days. Hypericum species were administered orally at a dose of 10, 30 or 100 mg/kg of body weight daily for 35 days. Body weight, food intake, anorexic effect and various biochemical parameters, such as serum glucose, lipid profile, alanine transaminase (ALT), aspartate transaminase (AST) and atherogenic index (AI), were assessed. Additionally, inhibitory lipase activity assay and forced swimming test were also carried out. KEY FINDINGS: Oral administration of H. silenoides and H. philonotis extracts resulted in a significant decrease in body weight and serum glucose levels in obese male Wistar rats. Treatment with aqueous extract of H. silenoides showed anorexic and antidepressant effects and also significantly (P < 0.05) decreased total cholesterol, triglycerides and high-density lipoprotein-cholesterol, while low-density lipoprotein-cholesterol, AI, AST and ALT were not changed. The dichloromethane extract of H. silenoides (half maximal inhibitory concentration (IC50 ) = 262.79 ± 0.09 µg/ml) and hexane extract of H. philonotis (IC50 = 162.60 ± 0.02 µg/ml) showed the most potent lipase inhibitory activity. CONCLUSION: Some H. silenoides and H. philonotis extracts showed a significant anti-obesity activity in cafeteria-diet-fed rats. This research provides the first scientific support for the use of the Hypericum genus for weight reduction in Mexican folk medicine.
Assuntos
Fármacos Antiobesidade/uso terapêutico , Etnofarmacologia , Hypericum/química , Hipolipemiantes/uso terapêutico , Obesidade/tratamento farmacológico , Componentes Aéreos da Planta/química , Extratos Vegetais/uso terapêutico , Animais , Fármacos Antiobesidade/administração & dosagem , Fármacos Antiobesidade/isolamento & purificação , Fármacos Antiobesidade/farmacologia , Antidepressivos/administração & dosagem , Antidepressivos/isolamento & purificação , Antidepressivos/farmacologia , Antidepressivos/uso terapêutico , Depressão/etiologia , Depressão/prevenção & controle , Relação Dose-Resposta a Droga , Inibidores Enzimáticos/administração & dosagem , Inibidores Enzimáticos/isolamento & purificação , Inibidores Enzimáticos/farmacologia , Inibidores Enzimáticos/uso terapêutico , Hiperglicemia/etiologia , Hiperglicemia/prevenção & controle , Hypericum/crescimento & desenvolvimento , Hiperlipidemias/etiologia , Hiperlipidemias/prevenção & controle , Hipoglicemiantes/administração & dosagem , Hipoglicemiantes/isolamento & purificação , Hipoglicemiantes/farmacologia , Hipoglicemiantes/uso terapêutico , Hipolipemiantes/administração & dosagem , Hipolipemiantes/isolamento & purificação , Hipolipemiantes/farmacologia , Lipase/antagonistas & inibidores , Masculino , Medicina Tradicional , México , Obesidade/sangue , Obesidade/fisiopatologia , Componentes Aéreos da Planta/crescimento & desenvolvimento , Extratos Vegetais/administração & dosagem , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/farmacologia , Distribuição Aleatória , Ratos , Ratos WistarRESUMO
CONTEXT: The genus Hypericum (Guttiferae) has received considerable scientific interest as a source of biologically active compounds. OBJECTIVE: The study determined the morphogenetic and ontogenetic variation in the main bioactive compounds of two Hypericum species, namely, Hypericum aviculariifolium subsp. depilatum var. depilatum (Freyn and Bornm.) Robson var. depilatum and Hypericum orientale L. through HPLC analyses of whole plants as well as individual plant parts (stems, leaves, and reproductive tissues). MATERIALS AND METHODS: The plant materials were harvested at five phenological stages: vegetative, floral budding, full flowering, fresh fruiting, and mature fruiting; dried at room temperature, then assayed for chemical content. RESULTS: In H. aviculariifolium, no kaempferol accumulation was observed and the highest level of hypericin, pseudohypericin, and quercitrin was reached at full flowering (0.71, 1.78, and 4.15 mg/g DW, respectively). Plants, harvested at floral budding produced the highest amount of rutin, hyperoside, and isoquercitrine (32.96, 2.42, 1.52 mg/g DW, respectively). H. orientale did not produce hypericin, pseudohypericin, or kaempferol. Rutin, hyperoside, and isoquercetine levels were the highest at floral development (1.76, 11.85, and 1.21 mg/g DW, respectively) and plants harvested at fresh fruiting produced the highest amount of quercitrine and quercetine (0.20 and 1.30 mg/g DW, respectively). DISCUSSION: For the first time, the chemical composition of the Turkish species of Hypericum was monitored during the course of ontogenesis to determine the ontogenetic and morphogenetic changes in chemical content. CONCLUSIONS: Plant material should be harvested during flower ontogenesis for medicinal purposes in which the content of many bioactive substances tested reached their highest level.