RESUMEN

ETHNOPHARMACOLOGICAL RELEVANCE: Pfaffia glomerata roots are widely used in Brazil to treat various pathological conditions, particularly psychological disorders. 20-hydroxyecdysone, a phytosteroid present in the plant, can promote greater body resistance against exogenous and endogenous stressors. The objective of this study was to evaluate the possible neuroprotective effect of a 20-hydroxyecdysone-enriched fraction (20E-EF), obtained from P. glomerata roots, in an acute murine stress model. MATERIAL AND METHODS: The 20E-EF was obtained by partitioning the methanol extract from P. glomerata roots with dichloromethane. Mice were treated by gavage with three doses of 20E-EF (3, 10, and 30 mg/kg) and parameters of stress, anxiety, and depression were evaluated. Biomarkers of oxidative stress (enzymes, antioxidant profile, and oxidized molecules) were evaluated in the cortex, striatum (basal ganglia), and hippocampus of animals treated with 30 mg/kg of 20E-EF. RESULTS: Mass spectrometry revealed that 20E was the main compound in the dichloromethane fraction. At a dose of 30 mg/kg, 20E-EF reduced stress, anxiety, and depression, while stimulating antioxidant enzymes (catalase, superoxide dismutase, and glutathione peroxidase), promoting antioxidant activity (antioxidant capacity, sulfhydryl groups, and reduced glutathione), and reducing oxidative markers (lipid peroxidation). In addition, 20E increased the concentration of NO in the striatum, possibly improving memory function and antioxidant activity. CONCLUSION: A 30 mg/kg dose of 20E-EF was able to reduce stress, anxiety, and depression, in addition to maintaining antioxidant defenses of the cortex and striatum. These findings open new perspectives for understanding the therapeutic properties of P. glomerata and the underlying mechanism(s).

Asunto(s)

Amaranthaceae , Ansiolíticos/farmacología , Antidepresivos/farmacología , Ansiedad/prevención & control , Conducta Animal/efectos de los fármacos , Encéfalo/efectos de los fármacos , Depresión/prevención & control , Ecdisterona/farmacología , Extractos Vegetales/farmacología , Raíces de Plantas , Estrés Psicológico/prevención & control , Amaranthaceae/química , Animales , Ansiolíticos/aislamiento & purificación , Antidepresivos/aislamiento & purificación , Antioxidantes/farmacología , Ansiedad/metabolismo , Ansiedad/fisiopatología , Ansiedad/psicología , Biomarcadores/metabolismo , Encéfalo/metabolismo , Encéfalo/fisiopatología , Depresión/metabolismo , Depresión/fisiopatología , Depresión/psicología , Modelos Animales de Enfermedad , Ecdisterona/aislamiento & purificación , Conducta Exploratoria/efectos de los fármacos , Peroxidación de Lípido/efectos de los fármacos , Masculino , Memoria/efectos de los fármacos , Ratones Endogámicos C57BL , Actividad Motora/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Extractos Vegetales/aislamiento & purificación , Raíces de Plantas/química , Estrés Psicológico/metabolismo , Estrés Psicológico/fisiopatología , Estrés Psicológico/psicología

RESUMEN

Pfaffia glomerata possesses potential pharmacological and medicinal properties, mainly owing to the secondary metabolite 20-hydroxyecdysone (20E). Increasing production of biomass and 20E is important for industrial purposes. This study aimed to evaluate the influence of irradiance on plant morphology and production of 20E in P. glomerata grown in vitro. Nodal segments of accessions 22 and 43 (Ac22 and Ac43) were inoculated in culture medium containing MS salts and vitamins. Cultures were maintained at 25 ± 2 °C under a 16-h photoperiod and subjected to irradiance treatments of 65, 130, and 200 µmol m-2 s-1 by fluorescent lamps. After 30 days, growth parameters, pigment content, stomatal density, in vitro photosynthesis, metabolites content, and morphoanatomy were assessed. Notably, Ac22 plants exhibited 10-fold higher 20E production when cultivated at 200 µmol m-2 s-1 than at 65 µmol m-2 s-1, evidencing the importance of light quantity for the accumulation of this metabolite. 20E production was twice as high in Ac22 as in Ac43 plants although both accessions responded positively to higher irradiance. Growth under 200 µmol m-2 s-1 stimulated photosynthesis and consequent biomass accumulation, but lowered carotenoids and anthocyanins. Furthermore, increasing irradiance enhanced the number of palisade and spongy parenchyma cells, enhancing the overall growth of P. glomerata. Graphical abstract.

Asunto(s)

Amaranthaceae/química , Fotosíntesis/genética , Técnicas In Vitro

RESUMEN

The combination of different colors from light-emitting diodes (LEDs) may influence growth and production of secondary metabolites in plants. In the present study, the effect of light quality on morphophysiology and content of 20-hydroxyecdysone (20E), a phytoecdysteroid, was evaluated in accessions of an endangered medicinal species, Pfaffia glomerata, grown in vitro. Two accessions (Ac22 and Ac43) were cultured in vitro under three different ratios of red (R) and blue (B) LEDs: (i) 1R:1B, (ii) 1R:3B, and (iii) 3R:1B. An equal ratio of red and blue light (1R:1B) increased biomass accumulation, anthocyanin content, and 20E production (by 30-40%). Moreover, 1R:1B treatment increased the size of vascular bundles and vessel elements, as well as strengthened xylem lignification and thickening of the cell wall of shoots. The 1R:3B treatment induced the highest photosynthetic and electron transport rates and enhanced the activity of oxidative stress-related enzymes. Total Chl content, Chl/Car ratio, and NPQ varied more by accession type than by light source. Spectral quality affected primary metabolism differently in each accession. Specifically, in Ac22 plants, fructose content was higher under 1R:1B and 1R:3B treatments, whereas starch accumulation was higher under 1R:3B, and sucrose under 3R:1B. In Ac43 plants, sugars were not influenced by light spectral quality, but starch content was higher under 3R:1B conditions. In conclusion, red and blue LEDs enhance biomass and 20E production in P. glomerata grown in vitro.

Asunto(s)

Amaranthaceae/efectos de la radiación , Ecdisterona/análisis , Luz , Amaranthaceae/crecimiento & desarrollo , Amaranthaceae/metabolismo , Antocianinas/análisis , Biomasa , Carotenoides/análisis , Catalasa/metabolismo , Clorofila/análisis , Cromatografía Líquida de Alta Presión , Hojas de la Planta/química , Proteínas de Plantas/metabolismo , Raíces de Plantas/química , Raíces de Plantas/crecimiento & desarrollo , Tallos de la Planta/química , Plantas Medicinales/crecimiento & desarrollo , Plantas Medicinales/metabolismo , Plantas Medicinales/efectos de la radiación , Almidón/metabolismo , Superóxido Dismutasa/metabolismo

RESUMEN

Somatic embryogenesis from explants from hermaphrodite papaya mother plants is an alternative for the production of true-to-type plants without the need for sexing. This study aimed to analyze hormonal and osmotic inducers in different somatic embryogenesis stages in the commercial hermaphrodite hybrid papaya UENF/Caliman 01. Leaf disks from in vitro shoots originated from ex vitro hermaphrodite plants were cultured in induction medium supplemented with different concentrations of 2,4-D (6, 9, 12, 15, and 18 µM) and 4-CPA (19, 22, 25, 28, and 31 µM). After 90 days, the formation of somatic embryos was verified. The 2,4-D induced the formation of light brown calli with low frequency (20%) of somatic embryogenesis. However, 4-CPA (25 µM) induced 96% of embryogenic calli, which were transferred to maturation medium (MM) and cultured for 30 days. The MM contained ABA (0.5 µM) and AC (15 g L-1) and produced 36.6 somatic embryos callus-1, mainly on cotyledonary stage. Cotyledonary embryos were transferred to germination medium supplemented with gibberellic acid (GA3) (0.0, 1.44, 2.88, and 4.32 µM), and the conversion into plantlets was enhanced with GA3 at 2.88 µM.

Asunto(s)

Carica/embriología , Carica/fisiología , Germinación/fisiología , Técnicas de Embriogénesis Somática de Plantas/métodos , Factores de Tiempo

RESUMEN

Ultraviolet-B (UV-B) radiation is an elicitor of secondary metabolites in plant tissue culture, but the effects on 20-hydroxyecdysone (20E) are still unclear. The 20E may show biotechnological, pharmacological, medical, and agrochemical applicability. Here, we use Pfaffia glomerata, a medically important species, to understand the impacts of UV-B radiation on their physiological performance, the expression of key genes involved in the 20E biosynthesis, and the 20E content. Two accessions (A22 and A43) of plants 20 days old grown in vitro were exposed to 0 (control), 2 (6.84 kJ m-2), and 4 (13.84 kJ m-2) h UV-B radiation for 20 consecutive days. Our data showed that UV-B reduced glucose concentration in A22 and A43 under 4 h of exposure (29 and 30%, respectively), while sucrose concentration increased (32 and 57%, respectively). UV-B also differentially impacted the accessions (A22 and A43), where the A22 under 4 h of UV-B had reduced total dry weight (8%) and electron transport rate (31%); in contrast, A43 did not change. Also, only A22 had increased POD activity under 4 h of UV-B (66%), as well as increased gene expression of the 20E pathway and the 20E content under 2 and 4 h of UV-B in leaves (28 and 21%, respectively) and roots (16 and 13%, respectively). This differential performance to UV-B can be explained by the contrasting anthocyanin contents. Notably, A43 displayed 56% more anthocyanin to the former, a possible defense against UV-B. In conclusion, UV-B radiation is a potential elicitor for increasing 20E content in P. glomerata grown in vitro.

Asunto(s)

Antocianinas/metabolismo , Panax/química , Brasil , Estrés Fisiológico , Rayos Ultravioleta

RESUMEN

- Salinity is a major threat to agriculture. However, depending on the concentration of soluble salts in soil, increased secondary metabolite levels can occur with no major damages to plant growth and development. The phytoecdysteroid (PE) 20-hydroxyecdysone (20E) is a secondary metabolite with biotechnological, medicinal, pharmaceutical and agrochemical applicability. Here, we characterize the responses (growth and physiology) of Pfaffia glomerata under different NaCl concentrations and examine the production of 20E as affected by salinity. Forty-day-old plants grown in greenhouse were exposed to 0, 120, 240, 360 or 480 mM of NaCl for 11 days. Moderate salinity (i.e., 120 mM of NaCl) led to increased 20E concentrations in leaves (47%) relative to the control with no significant effect on photosynthesis and biomass accumulation, thus allowing improved 20E contents on a per whole-plant basis. In contrast, plants under high salinity (i.e., 240-480 mM of NaCl) displayed similar 20E concentrations in leaves compared to the control, but with marked impairments to biomass accumulation and photosynthetic performance (coupled with decreased sucrose and starch levels) in parallel to nutritional imbalance. High salinity also strongly increased salicylic acid levels, antioxidant enzyme activities, and osmoregulatory status. Regardless of stress severity, 20E production was accompanied by the upregulation of Spook and Phantom genes. Our findings suggest that P. glomerata cultivation in moderate salinity soils can be considered as a suitable agricultural option to increase 20E levels, since metabolic and structural complexity that makes its artificial synthesis very difficult.

Asunto(s)

Panax/metabolismo , Cloruro de Sodio/farmacología , Biomasa , Ecdisterona/metabolismo , Fotosíntesis/efectos de los fármacos , Salinidad

RESUMEN

This study evaluated the effect of osmoregulators and carbohydrates on the maturation and germination of somatic embryos of papaya 'Golden THB'. Cotyledon explants from papaya seedlings germinated in vitro on basal MS medium were cultured on somatic embryogenesis induction medium (IM) containing MS salts, myo-inositol, sucrose, agar and p-chlorophenoxyacetic acid. After 50 days, embryogenic calli were transferred onto maturation media (MM) for 45 additional days. For experiment 1, a MS-based medium supplemented with abscisic acid, activated charcoal and concentrations of PEG 6000 (0; 40; 50; 60 and 70 g L-1) was used, whereas for experiment 2 malt extract concentrations (0; 0.1; 0.2; 0.3 and 0.4 g L-1) were assessed. The normal cotyledonary somatic embryos produced in experiment 2 were transferred to the germination medium (GM). The GM consisted of full-strength MS medium, sucrose, agar and was supplemented with myo-inositol at varying concentrations (0; 0.275; 0.55 and 0.825 mM). The PEG concentrations tested impaired the maturation of 'Golden THB' papaya somatic embryos. The MM, supplemented with malt extract at 0.153 g L-1, promoted the greatest development of normal somatic embryos (18.28 SE calli-1), that is, two cotyledonary leaves produced 36.56 SE calli-1. The supplementation with 0.45 mM myo-inositol provided the highest germination percentage (47.42%) and conversion to emblings.

Asunto(s)

Ácido Abscísico/farmacología , Carbohidratos/farmacología , Carica/efectos de los fármacos , Germinación/efectos de los fármacos , Osmorregulación , Reguladores del Crecimiento de las Plantas/farmacología , Brotes de la Planta/efectos de los fármacos , Técnicas de Embriogénesis Somática de Plantas/métodos , Polímeros/farmacología , Carica/crecimiento & desarrollo , Brotes de la Planta/crecimiento & desarrollo

RESUMEN

This study aimed to evaluate the herbicidal activity of picloram on the biomass of the remediation plants Eleusine coracana and Panicum maximum after cultivation in a soil contaminated with this herbicide. These species were grown in three soils, differentiated based on texture (clayish, middle, and sandy, with 460, 250, and 40 g kg(-1) of the clay, respectively), previously contaminated with picloram (0, 80, and 160 g ha(-1)). After 90 days, the plants were harvested and an extract was produced by maceration of leaves and stems of these plants. It was applied to pots containing washed sand, comprising a bioassay in a growth chamber using soybean as a bioindicator for picloram. Soil and plant samples were analyzed by HPLC. The results showed the presence of picloram or metabolites with herbicidal activity in the shoots of E. coracana and P. maximum at phytotoxic levels with regard to soybean plants, indicating that they work only as phytoextractors and that the presence of straw on the soil surface can promote recontamination within the area. It is not recommended to cultivate species susceptible to picloram in areas where it was reported remediation by E. indica and P. maximum and still present residues of these species.

Asunto(s)

Eleusine/metabolismo , Restauración y Remediación Ambiental/métodos , Herbicidas/metabolismo , Panicum/metabolismo , Picloram/metabolismo , Contaminantes del Suelo/metabolismo , Biodegradación Ambiental , Eleusine/química , Restauración y Remediación Ambiental/instrumentación , Herbicidas/análisis , Panicum/química , Picloram/análisis , Tallos de la Planta/química , Tallos de la Planta/metabolismo , Contaminantes del Suelo/análisis

RESUMEN

Este trabalho teve como objetivo selecionar espécies com potencial para a fitorremediação de solos contaminados com o herbicida sulfentrazone. Foram avaliadas oito espécies: Arachis pintoi, Eleusine coracana, Crotalaria spectabilis, Crotalaria ochroleuca, Cajanus cajan, Leucaena leucocephala, Stizolobium cinereum e Raphanus sativus. O experimento foi instalado em casa de vegetação, com a utilização de vasos com capacidade para 6 dm3 preenchidos com porções de solo coletadas na profundidade de 0-20 cm. O delineamento experimental foi o de blocos casualizados em esquema fatorial 8 × 5, com quatro repetições, composto pela combinação entre as espécies e cinco doses do sulfentrazone (0, 200, 400, 800 e 1.600 g ha-1). Foram avaliadas a fitotoxicidade do herbicida, a altura de plantas e a massa da matéria seca da parte aérea e de raízes. As espécies Cajanus cajan e Leucaena leucocephala apresentaram maior tolerância ao sulfentrazone até a dose de 400 g ha-1, mostrando menores sintomas de fitotoxicidade e menores reduções na altura de plantas e no acúmulo de matéria seca, tanto na parte aérea como nas raízes, em relação ao tratamento controle, indicando, com isso, potencial de utilização para posteriores estudos de fitorremediação de sulfentrazone em solo.

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This paper aimed to select species with potential for phytoremediation of soils contaminated with the herbicide sulfentrazone. Eight species were evaluated: Arachis pintoi, Eleusine coracana, Crotalaria spectabilis, Crotalaria ochroleuca, Cajanus cajan, Leucaena leucocephala, Stizolobium cinereum, and Raphanus sativus. The experiment was set-up inside a greenhouse, using pots with a capacity of 6dm3 filled with soil samples collected at a depth of 0-20cm. The experimental design was arranged into randomised blocks in a factorial scheme (8 × 5) with four replications, which consisted of the combination between the species and five doses of sulfentrazone (0, 200, 400, 800, and 1,600g ha-1). The herbicide phytotoxicity, plant heights, and dry masses of shoots and roots were evaluated. The species Cajanus cajan and Leucaena leucocephala had a higher tolerance to sulfentrazone up to a dose of 400g ha-1, showing minor symptoms of phytotoxicity and smaller decreases in plant heights and in dry matter accumulation, both in the shoots and roots, when compared to the control treatment, indicating, thus, the potential to be used for further studies on phytoremediation of sulfentrazone in soil.

RESUMEN

Este trabalho teve como objetivo selecionar espécies com potencial para a fitorremediação de solos contaminados com o herbicida sulfentrazone. Foram avaliadas oito espécies: Arachis pintoi, Eleusine coracana, Crotalaria spectabilis, Crotalaria ochroleuca, Cajanus cajan, Leucaena leucocephala, Stizolobium cinereum e Raphanus sativus. O experimento foi instalado em casa de vegetação, com a utilização de vasos com capacidade para 6 dm3 preenchidos com porções de solo coletadas na profundidade de 0-20 cm. O delineamento experimental foi o de blocos casualizados em esquema fatorial 8 × 5, com quatro repetições, composto pela combinação entre as espécies e cinco doses do sulfentrazone (0, 200, 400, 800 e 1.600 g ha-1). Foram avaliadas a fitotoxicidade do herbicida, a altura de plantas e a massa da matéria seca da parte aérea e de raízes. As espécies Cajanus cajan e Leucaena leucocephala apresentaram maior tolerância ao sulfentrazone até a dose de 400 g ha-1, mostrando menores sintomas de fitotoxicidade e menores reduções na altura de plantas e no acúmulo de matéria seca, tanto na parte aérea como nas raízes, em relação ao tratamento controle, indicando, com isso, potencial de utilização para posteriores estudos de fitorremediação de sulfentrazone em solo.

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This paper aimed to select species with potential for phytoremediation of soils contaminated with the herbicide sulfentrazone. Eight species were evaluated: Arachis pintoi, Eleusine coracana, Crotalaria spectabilis, Crotalaria ochroleuca, Cajanus cajan, Leucaena leucocephala, Stizolobium cinereum, and Raphanus sativus. The experiment was set-up inside a greenhouse, using pots with a capacity of 6dm3 filled with soil samples collected at a depth of 0-20cm. The experimental design was arranged into randomised blocks in a factorial scheme (8 × 5) with four replications, which consisted of the combination between the species and five doses of sulfentrazone (0, 200, 400, 800, and 1,600g ha-1). The herbicide phytotoxicity, plant heights, and dry masses of shoots and roots were evaluated. The species Cajanus cajan and Leucaena leucocephala had a higher tolerance to sulfentrazone up to a dose of 400g ha-1, showing minor symptoms of phytotoxicity and smaller decreases in plant heights and in dry matter accumulation, both in the shoots and roots, when compared to the control treatment, indicating, thus, the potential to be used for further studies on phytoremediation of sulfentrazone in soil.