Search

The Pfaffia glomerata, a plant popularly called Brazilian ginseng, is widely used in Brazil for the treatment of various pathologies, including those associated with the Central Nervous System. 20-hydroxyecdysone (20E), a phytosteroid present in this plant, can promote adaptogenic effects in the organism, providing greater body resistance to stressors. This study aimed to evaluate the phytochemical composition and the anticholinesterase, antioxidant, and antiglycation effects of extracts and fractions of aerial parts and roots of P. glomerata, also analyzing their possible cytotoxic effects. The fractions were obtained by partitioning methanol extracts from the aerial part and roots of P. glomerata with hexane, dichloromethane, ethyl acetate, n-butanol, and water. The samples were initially tested in anticholinesterase, antioxidant, and antiglycation assays, and the most promising samples were submitted for cytotoxicity and chromatographic analyses. Mass spectrometry and chromatography methods revealed that 20E was the main compound in the dichloromethane fractions, there being 35% more 20E in the aerial part (APD) than in the roots (RD). Added to the higher concentration of 20E, the APD fraction also presented more promising results than the RD fraction in anticholinesterase and antioxidant analyses, indicating that their effects may be related to the concentration of 20E. These same fractions showed no hemolytic effects but were cytotoxic in high concentrations. These new findings contribute to scientific information about P. glomerata and open more perspectives for the understanding of its therapeutic properties, allowing the association of biological activity with the presence of 20E. [ABSTRACT FROM AUTHOR]

The World Health Organization (WHO) has prioritized developing new drugs against specific bacteria and fungi, such as Enterobacteriaceae and Candida spp. While Pfaffia paniculata is commonly called the "cure-everything", its scientifically proven benefits are limited to anti-inflammatory and antioxidant actions. Therefore, this study aims to determine the spectrum of antimicrobial activity of Pfaffia paniculata and assess its cytotoxicity. Thus, broth microdilution test was conducted according to the CLSI M7-A9 and M27-A3 reference methods. After screening, microbial species with minimum inhibitory concentration (MIC) values were selected for biofilm tests. These tests evaluated biomass using the crystal violet (CV) test, metabolic activity using the MTT assay, and structural analysis via Scanning Electron Microscopy (SEM). Cytotoxicity was evaluated in human gingival fibroblasts (FMM-1). There were reductions of 29.4 and 42.7% in CV and MTT assays for Candida spp. biofilm. S. mutans and P. aeruginosa biofilms showed a decrease of 15.7 and 28.6%, respectively. Cell viability tests indicated 55.1, 56.9, and 65.5% of viability after contact with 1.93, 0.96, and 0.48 mg/mL of the extract, respectively. The P. paniculata extract showed antimicrobial action, displayed MIC values, and antibiofilm action on P. aeruginosa, S. mutans, and C. albicans. The cytotoxicity on the FMM-1 cell line was dose-dependent. Therefore, P. paniculata extract holds significant potential for developing new drugs. [ABSTRACT FROM AUTHOR]

Summary: This study aimed to evaluate the convective drying of a by‐product obtained by hydrodistillation of Pfaffia glomerata roots (PGB) and investigate the utilisation of the resulting powder in the preparation of cereal bars. Drying at 80 °C for 120 min was the selected condition for PGB due to the higher levels of phenolic acids (gallic, coumaric and ferulic) catechin and higher antioxidant activity than drying at 60 °C for 180 min, and, better technological properties and higher β‐ecdysone content than drying at 100 °C for 100 min. There was an increase in fibre content (110%), antioxidant compounds (97%) and compressive strength (377%) and a decrease in lipid content (31%) when the PGB content of cereal bars was increased from 0% to 9%. Colour parameters, purchase intention and most sensory parameters were similar among cereal bar formulations, indicating that PGB is a promising ingredient for the nutritional enrichment of cereal bars. [ABSTRACT FROM AUTHOR]

The World Health Organization (WHO) has prioritized developing new drugs against specific bacteria and fungi, such as Enterobacteriaceae and Candida spp. While Pfaffia paniculata is commonly called the “cure-everything”, its scientifically proven benefits are limited to anti-inflammatory and antioxidant actions. Therefore, this study aims to determine the spectrum of antimicrobial activity of Pfaffia paniculata and assess its cytotoxicity. Thus, broth microdilution test was conducted according to the CLSI M7-A9 and M27-A3 reference methods. After screening, microbial species with minimum inhibitory concentration (MIC) values were selected for biofilm tests. These tests evaluated biomass using the crystal violet (CV) test, metabolic activity using the MTT assay, and structural analysis via Scanning Electron Microscopy (SEM). Cytotoxicity was evaluated in human gingival fibroblasts (FMM-1). There were reductions of 29.4 and 42.7% in CV and MTT assays for Candida spp. biofilm. S. mutans and P. aeruginosa biofilms showed a decrease of 15.7 and 28.6%, respectively. Cell viability tests indicated 55.1, 56.9, and 65.5% of viability after contact with 1.93, 0.96, and 0.48 mg/mL of the extract, respectively. The P. paniculata extract showed antimicrobial action, displayed MIC values, and antibiofilm action on P. aeruginosa, S. mutans, and C. albicans. The cytotoxicity on the FMM-1 cell line was dose-dependent. Therefore, P. paniculata extract holds significant potential for developing new drugs.

Pfaffia glomerata (Spreng.) Pedersen, Amaranthaceae, is found in South America, mainly in Brazil, where it is considered a species of great medicinal interest owing to its popular use as a tonic, aphrodisiac, anti-inflammatory, and analgesic. These properties can be attributed to the presence of the phytosteroid, 20-Hydroxyecdysone (β-ecdysone), the main compound found in its roots. This review aims to provide information about the botanical characteristics, ethnomedicinal uses, the phytochemistry, the biological activities, and the biotechnology of P. glomerata , an important species to local communities and groups researching medicinal plants of South America. The information available on P. glomerata was collected from scientific databases (ScienceDirect, PubMed/MEDLINE, SciELO, and Scopus) until June 7, 2023, using the search terms " Pfaffia glomerata ", " Pfaffia glomerata (Spreng.) Pedersen", and "Brazilian ginseng". The review includes studies that evaluated the botanical, ethnopharmacological, and phytochemical aspects, biological properties, nutraceutical uses, and the application of biotechnology for improving the biosynthesis of metabolites of interest. A total of 207 studies were identified, with 81 articles read in full. Seventy-six studies were included for qualitative synthesis. Overall, 40 compounds belonging to different classes are presented in this review, including ecdysteroids, triterpenes, saponins, flavonoids, anthraquinones, tannins, coumarins, alkaloids, and polysaccharides. Among them, flavonoids, anthraquinones, tannins, coumarins, and alkaloids were only putatively identified. β-Ecdysone, triterpenes, saponins, and polysaccharides are the chemical components most frequently identified and isolated from P. glomerata and possibly responsible for ethnopharmacological use and the biological activities of this species, with important in vitro and in vivo activities, such as anti-inflammatory, antidepressant, aphrodisiac, analgesic, gastroprotective, antioxidant, and prebiotic. This review summarizes discussions about the P. glomerata species, highlighting its ethnopharmacological, chemical, biotechnological, and nutraceutical importance. New scientific studies on this species are encouraged in the search for new therapeutic molecules with pharmaceutical potential and nutraceutical applications. [Display omitted] [ABSTRACT FROM AUTHOR]

[Display omitted] • β-ecdysone was obtained from an in vitro cultured of Pfaffia glomerata. • The roller bottle system was favourable to the adventitious root of P. glomerata. • Presence of β-ecdysone in culture of P. glomerata means adaptation to the system. • Highest accumulation of β-ecdysone occurred after 8 weeks in a roller bottle system. β-ecdysone is the main compound of commercial interest produced by the roots of Pfaffia glomerata (Spreng.) Pedersen, and is used in herbal medicines and dietary supplements. In-vitro root culture facilitates the continuous production of high-quality compounds, and uses sustainable production techniques. Adventitious roots of P. glomerata were cultivated in a roller bottle system and an orbital gyratory system, both in the absence of light. The quantification of β-ecdysone by Liquid Chromatography-Mass Spectrometry/Mass Spectrometry (LC–MS/MS) revealed that the highest accumulation of metabolites occurred after 8 weeks in the roller bottle culture system, wherein 1.81 μg of β-ecdysone was recovered from 30 mg of dry root. These results suggest that adventitious root culture in-vitro may be an alternative strategy for β-ecdysone production, and demonstrates the potential use of bioreactors for practical applications. [ABSTRACT FROM AUTHOR]

Pfaffia glomerata has potential pharmacological and medicinal properties due to the production of a secondary metabolite known as the phytoecdysteroid 20-hydroxyecdysone (20E). There have been increasing efforts for massive in vitro propagation of Pfaffia plants due to high extractivism and overharvesting of this species. Research on the species has shown that photoautotrophic cultivation can improve the production of 20E. In addition, other abiotic factors such as the formulations of culture media can influence the morphophysiological behavior of the plants in vitro. Therefore, the objective of this study was to analyze the morphological and physiological performances of P. glomerata plants in different formulations of culture media, under photoautotrophic and photomixotrophic propagation conditions. Six medium formulations, the Driver and Kuniyuki medium (DKW), Correia et al. medium (JADS), Murashige and Skoog medium (MS), Quoirin and Lepoivre medium (QL), Rugini medium (OM), and Woody Plant medium (WPM), all supplemented with DKW vitamins, 100 mg L−1 myo-inositol, 6.5 g L−1 agar, and with or without 3% (w/v) sucrose, were evaluated. Cultures were maintained at 25 ± 2°C, with a 16 h-photoperiod under 60 μmol m−2 s−1 of irradiance under a fluorescent lamp for 50 d. Results showed that the presence or absence of sucrose, and the different nutritional formulations influenced growth, photosynthetic pigment content, endogenous levels of sugars, leaf morphology, levels of 20E, and transport of water and minerals in P. glomerata. Notably, OM, DKW, QL, and WPM media promoted higher production of 20E under photomixotrophic growth conditions. [ABSTRACT FROM AUTHOR]

The determination of the best reference gene is essential to improve and guarantee the accuracy of the qPCR technique. Thus, the objective of this work was to evaluate the normalization genes efficiency for qRT-PCR studies of Pfaffia glomerata, a species with marked medicinal and industrial interests due to the production of the phytoecdysteroid 20-hydroxyecdysone (20-E). We have selected four candidates as reference genes in P. glomerata: elongation factor-1α (EF-1α), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), spectrin-like (SPT) and α-tubulin (TUA), and tested their expression stabilities as affected by abiotic factors (salt stress, drought stress, irradiance, photoperiod and CO2 enrichment), using the following methods: NormFinder, geNorm, BestKeeper, ΔCt and RefFinder. Also, the Phantom gene, which belongs to the 20-E biosynthesis pathway, was targeted to validate the most stable reference gene for each abiotic factor. The GAPDH was the most stable gene under all assessed abiotic factors, as well as the most stable when data from all experiments were taken together, which was confirmed by all the software used. The use of more or less stable reference genes for normalization significantly changes the interpretation of the qPCR data, evidencing the importance of choosing the most appropriate housekeeping gene for each expression assay. Based on our results, we recommend GAPDH to be used for normalization of qPCR expression data in P. glomerata in diverse abiotic conditions. This work is the first report on the validation of reference genes in P. glomerata and will be fundamental for further gene expression studies in this important medicinal species. Key message: GAPDH is the most stable gene to be used for the normalization in qPCR analyzes in [Pfaffia glomerata (Spreng.) Pedersen] under the different abiotic factors. [ABSTRACT FROM AUTHOR]

Pfaffia glomerata is a medically important species because it produces the phytoecdysteroid 20-hydroxyecdysone (20-E). However, there has been no ready-to-use transcriptome data available in the literature for this plant. Here, we present de novo transcriptome sequencing of RNA from P. glomerata in order to investigate the 20-E production as well as to understand the biochemical pathway of secondary metabolites in this non-model species. We then analyze the effect of photoautotrophy on the production of 20-E genes phylogenetically identified followed by expression analysis. For this, total messenger RNA (mRNA) from leaves, stems, roots, and flowers was used to construct indexed mRNA libraries. Based on the similarity searches against plant non-redundant protein database, gene ontology, and eukaryotic orthologous groups, 164,439 transcripts were annotated. In addition, the effect of photoautotrophy in two genes putatively involved in the 20-E synthesis pathway was analyzed. The Phantom gene (CYP76C), a precursor of the route, showed increased expression in P. glomerata plants cultured under photoautotrophic conditions. This was accompanied by increased production of this metabolite indicating a putative involvement in 20-E synthesis. This work reveals that several genes in the P. glomerata transcriptome are related to secondary metabolism and stresses, that genes of the P450 family participate in the 20-E biosynthesis route, and that plants cultured under photoautotrophic conditions promote an upregulated Phantom gene and enhance the productivity of 20-E. The data will be used for future investigations of the 20-E synthesis pathway in P. glomerata while offering a better understanding of the metabolism of the species. [ABSTRACT FROM AUTHOR]

The production of plant seedlings has traditionally used polyethylene bags, which are thrown out in the soil or burned after transplant because the large amount of organic material attached to the bags makes recycling difficult. Additionally, when a seedling is taken from the bag for transplant, there is the risk of root damage, which compromises the plant’s development. In this study, we developed biodegradable bags to be used in seedling production, and we verify their influence on the development of Brazilian ginseng (Pfaffia glomerata (Spreng) Pedersen), when the plant is planted without being removed from the bag. Both black and white biodegradable bags remained intact throughout the seedling production period (60 days). After being transplanted into containers (240 days), they were completely biodegraded, and there was no significant difference between the dry mass of these plants and that of plants that were transplanted without the bags. The plants that were cultivated without being removed from the polyethylene bags had root development difficulties, and the wrapping showed no signs of degradation. The use of biodegradable films is an alternative for the production of bags for seedlings, as these can then be transplanted directly into the soil without removing the bag, reducing the risk of damage to the roots during the moment of transplant.

Tendo como objetivo avaliar a eficiência de absorção, translocação e uso de nitrogênio (N) e fósforo (P) pela Pfaffia glomerata, foi conduzido o presente trabalho em casa de vegetação no delineamento inteiramente casualizado com três repetições. Os tratamentos foram dispostos em esquema fatorial 5 x 5, sendo cinco doses de N (0, 125, 250, 375 e 500 mg vaso-1) e cinco doses de P (P2O5) (0, 125, 250, 375 e 500 mg vaso-1). Os resultados evidenciaram que a eficiência de absorção de N e P foi influenciada pelas doses de N e P utilizadas no experimento apresentando interação entre os fatores em estudo. A eficiência de translocação do N foi consideravelmente menor que a de P, provavelmente pela maior concentração de compostos nitrogenados nas raízes em detrimento da parte aérea. Observou-se maior eficiência de uso do P devido à menor concentração desse elemento na planta. A absorção de N e de P foi influenciada pela dose desses elementos no solo, sendo possível a estimativa das doses desses nutrientes que proporcionem maiores eficiências de absorção, translocação, uso e teor total.Aimed at evaluating the efficiency of nitrogen (N) and phosphorus (P) uptake, translocation and use by Pfaffia glomerata, the present study was conducted in greenhouse, in completely randomized design with three replicates. Treatments were displayed in 5 x 5 factorial design, with five N levels (0, 125, 250, 375 and 500 mg pot-1) and five P (P2O5) levels (0, 125, 250, 375 and 500 mg pot-1). Results evidenced that N and P uptake efficiency was influenced by the N and P levels used in the experiment, showing interaction among the studied factors. N translocation efficiency was considerably lower than that of P, probably due to the lower concentration of nitrogenous compounds in the roots than in the shoot. P use efficiency was higher due to the lower concentration of this element in the plant. N and P uptake was influenced by the level of these elements in the soil, being possible to estimate the levels of these nutrients which provide higher uptake, translocation and use efficiency besides total content.

Objetivou-se com este trabalho avaliar o efeito da incorporação ao solo de cama-de-frango e de fósforo sobre o crescimento e a produção da fáfia. Os tratamentos foram cinco doses de cama-de-frango semidecomposta (0, 5, 10, 15 e 20 t ha-1), com e sem o uso de fósforo (200 kg ha-1 de P2O5, na forma de superfosfato triplo), arranjados em esquema fatorial 5 x 2, no delineamento experimental blocos casualizados, com quatro repetições. As alturas das plantas variaram em função das doses da cama-de-frango, sendo maiores (138,50 cm) sob 20 t ha-1. As produções de massas frescas e secas de folhas, caules e raízes foram influenciadas significativamente pela interação cama-de-frango e fósforo. As maiores produções de massa fresca e seca de folhas (597 kg ha-1 e 100 kg ha-1, respectivamente) foram obtidas com a dose de 20 t ha-1 de cama-de-frango, sem o uso de fósforo. Para caules, as maiores produções de massa fresca e seca (7.757 kg ha-1 e 2.633 kg ha-1, respectivamente) foram obtidas com a dose de 20 t ha-1 de cama-de-frango, com o uso de fósforo. As maiores produções de massa fresca e seca de raízes (9.065 kg ha-1 e 377 kg ha-1, respectivamente) foram obtidas com a dose de 20 t ha-1 de cama-de-frango, sem o uso de fósforo. Os teores médios de N, P e K das folhas foram de 8,60; 0,88 e 4,46 g kg-1, respectivamente. Os teores de Ca e Mg foram máximos (7,56 e 5,12 g kg-1, respectivamente) com o uso de 20 t ha-1 de cama-de-frango, independente do uso de P. Nas raízes os teores médios de N, P e Mg foram de 2,74; 0,27 e 0,96 g kg-1, respectivamente. O teor de K reduziu linearmente com as doses de cama-de-frango e o de Ca foi máximo com o uso de 20 t ha-1. Para se obter maiores produções de massas frescas e secas de folhas e raízes, as plantas de fáfia devem ser cultivadas com cama-de-frango (20 t ha-1) incorporada ao solo, sem necessidade de adição de fósforo.The aimed of this study was to evaluate the effect of of chicken manure and phosphorus incorporated to the soil on growth and yield of fáfia. Treatments were five doses of chicken manure (0, 5, 10, 15 and 20 t ha-1), with and without the use of phosphorus (200 kg ha-1 of P2O5, in triple superphosphate form), arranged in 5 x 2 factorial scheme in randomized block design with four replications. Plant heights varied as a function of chicken manure doses, which the highest (138.50 cm) under 20 t ha-1. Yields of fresh and dried weight of leaves, stems and roots were influenced significantly by the interaction chicken manure x phosphorus. The highest yields of fresh and dried leaves (597 kg ha-1 and 100 kg ha-1, respectively) were obtained with the dose of 20 t ha-1 of chicken manure, without the use of phosphorus. For stems, the highest yields of fresh and dried weight (7.757 kg ha-1 and 2.633 kg ha-1, respectively) were obtained with the dose of 20 t ha-1 of chicken manure, with the use of phosphorus. The highest yields of fresh and dried weight of roots (9.065 kg ha-1 and 377 kg ha-1, respectively) were obtained with the dose of 20 t ha-1 of chicken manure, without the use of phosphorus. Average contents of N, P and K of leaves were 8.60; 0.88 and 4.46 g kg-1, respectively. Contents of Ca and Mg were maximum (7.56 and 5.12 g kg-1, respectively) with the use of 20 t ha-1 of chicken manure, independent on the use of phosphorus. In roots, average contents of N, P and Mg were 2.74, 0.27 and 0.96 g kg-1, respectively. K content reduced linearly with chicken manure doses and of Ca was maximum with the use of 20 t ha-1. For obtaining the highest yields of fresh and dried weight of leaves and roots, the fáfia plants must be grown with chicken manure (20 t ha-1) incorporated to the soil within the use of phosphorus.

Pfaffia glomerata ocorre em vários estados do Brasil e países limítrofes da região Sul às margens de rios e nas orlas das matas de galerias, é espécie hidrófita e heliófita. As raízes de espécies do gênero Pfaffia são usadas na medicina popular brasileira, especialmente como tônico, afrodisíaco e no controle do diabete. O objetivo deste trabalho foi estabelecer um banco de germoplasma in vitro de Pfaffia glomerata. O experimento em delineamento inteiramente casualizado foi conduzido com seis tratamentos: 1) MS + 2% de sacarose + 4% de sorbitol; 2) MS/2 + 2% de sacarose + 4% de sorbitol; 3) MS + 2% de sacarose + 4% de sorbitol + 2 mg L-1 de pantotenato de cálcio; 4) MS/2 + 2% de sacarose + 4% de sorbitol + 2 mg L-1de pantotenato de cálcio; 5) MS + 2% de sacarose + 3% de manitol + 2 mg L-1de pantotenato de cálcio; 6) MS/2 + 2% de sacarose + 3% de manitol + 2 mg L-1de pantotenato de cálcio. Os resultados obtidos foram submetidos à análise de variância e ao teste de separação de médias de Scott Knott. Os tratamentos um, três e quatro apresentaram, significativamente, o maior número de segmentos nodais por haste, quando comparados com os tratamentos dois, cinco e seis. O tratamento dois foi o mais indicado para a conservação in vitro da espécie por ter promovido menor crescimento das plantas (altura de 3,1±1,9 cm), alto índice de sobrevivência, 100% de explantes com brotação e o maior número de brotos por explante, após seis meses de cultivo. Todas as plântulas produziram raízes e não houve formação de calos, também não ocorreu hiperhidricidade nos tratamentos avaliados. As plantas aclimatizadas apresentaram 100% de sobrevivência no ambiente ex vitro. A manutenção de acessos de P. glomerata no banco de germoplasma in vitro é viável tanto do ponto de vista da conservação quanto economicamente.Pfaffia glomerata occurs in several states of Brazil and its neighboring countries in the south region at riverbanks and gallery forests. It is a hydrophyte and heliophyte species. The roots of the genus Pfaffia are used in Brazilian folk medicine especially as tonic, aphrodisiac and to control diabetes. The aim of this work was to establish an in vitro germplasm bank for Pfaffia glomerata. The experiment was carried out in completely randomized design with six treatments: 1) DM + 2% sucrose + 4% sorbitol; 2) DM/2 + 2% sucrose + 4% sorbitol; 3) DM + 2% sucrose + 4% sorbitol + 2 mg L-1 calcium pantothenate; 4) DM/2 + 2% sucrose + 4% sorbitol + 2 mg L-1 calcium pantothenate; 5) DM + 2% sucrose + 3% mannitol + 2 mg L-1 calcium pantothenate; 6) DM/2 + 2% sucrose + 3% mannitol + 2 mg L-1 calcium pantothenate. Results were subjected to analysis of variance and Scott Knott test for mean grouping. Treatments 1, 3 and 4 had a significantly larger number of nodal segments per stem, compared to Treatments 2, 5 and 6. Treatment 2 was the most appropriate for the in vitro conservation of this species since it led to the lowest growth (3.1±1.9 cm height), high survival rate, 100% explants with sprouting, and the largest number of sprouts per explant after six months of culture. All seedlings produced root and showed no formation of calluses or hyperhydricidity under the evaluated treatments. Acclimatized plants showed 100% survival in the ex vitro environment. Maintaining P. glomerata accessions in an in vitro germplasm bank is viable both economically and for conservation.

Conduziu-se este trabalho, com o objetivo de avaliar a propagação sexuada e assexuada de Pfaffia glomerata (ginseng brasileiro). As sementes foram pré-embebidas por 24 horas em KNO3 1%; ácido giberélico 50, 100 e 200 mg L-1 ; emágua e as sementes que não receberam tratamento serviram como testemunha. A semeadura foi em placas de petri sobre duas folhas de papel de filtro e a incubação a 25ºC, sob iluminação constante e na ausência de luz. As estacas com º15 cm foram imersas durante 6 horas em IAA 250 e IBA 500 mg L-1, ácido bórico 1% e em água e plantadas em embalagem plástica, contendo terra+areia (1:1). O primeiro experimento foi conduzido em delineamento inteiramente casualizado em esquema fatorial 2 (níveis de luz) x 6 (tratamento de pré-embebição) em 4 repetições de 20 sementes. O segundo em DIC com 5 tratamentos, 4 repetições de 15 estacas por repetição. As sementes de ginseng brasileiro necessitam de tratamento de pré-embebição para acelerar a germinação, sendo a imersão em KNO3 o método mais eficiente, proporcionando 63% de germinação. Apresentam fotoblastismo neutro, com germinação média de 45%. As estacas de ginseng brasileiro imersas apenas em água apresentaram 100% de enraizamento, maior número de brotos (2,5) e maior comprimento de raiz (8,6 cm). A espécie apresenta potencial para ambos os métodos de propagação.The objective of this study was to evaluate sexual and asexual propagation of Pfaffia glomerata (Brazilian Ginseng). The seeds were pre-soaked for 24 hours in KNO3 1% , gibberellic acid 50, 100, 200 mg L-1 , and in water. Untreated seeds served as control. Sowing was accomplished in petri dishes on 2 sheets of filter paper, at 25ºC incubation, under constant illumination, and in the absence of light. The cuttings with º1,5 cm were immersed for 6 hours in IAA 250 and 500mg L-1, boric acid1% ,and in water. Soon after they were planted in plastic containers with soil + sand (1:1). The first experiment was in factorial design 2 (light levels) x 6 (pre-soak treatment) in 4 repetitions of 20 seeds per repetition. The second in ERD in 5 treatments, 4 repetitions of 15 cuttings per repetition. Brazilian Ginseng seeds need pre-soak treatment to accelerate germination. We found immersion in KNO3 the most efficient method, providing 63% germination. They presented neuter photoblastism with an average germination of 45%. The Brazilian Ginseng cuttings immersed in water presented 100% rooting, a higher number of sprouts (2,5), and greater length (8,6 cm). The species presents potential for both propagation methods.

Pfaffia glomerata (Spreng.) Pedersen, conhecida como ginseng brasileiro, é uma planta extensivamente usada na medicina popular em decorrência de possuir propriedades fitoterápicas. O objetivo deste trabalho foi verificar a presença de β-ecdisona nas raízes e partes aéreas de plantas in vivo e in vitro provenientes de dois acessos (BRA e JB-UFSM) de Pfaffia glomerata através da cromatografia em camada delgada (CCD). Nas plantas in vivo, as manchas cromatográficas demonstraram que a β-ecdisona está presente na raiz e na parte aérea. Os acessos não apresentaram diferenças em relação ao perfil de manchas cromatográficas. A análise em CCD não detectou a presença de β-ecdisona nas plantas cultivadas in vitro.Pfaffia glomerata (Spreng.) Pedersen, known as Brazilian ginseng, is a plant extensively used in folk medicine due to its phytotherapic characteristics. The aim of this study was to evaluate the presence of β-ecdysone in the roots and shoots of in vivo and in vitro cultured plants of two sources (BRA and JB-UFSM) of Pfaffia glomerata using Thin-Layer Chromatography (TLC). For in vivo cultured plants, TLC plates showed that β-ecdysone was present in the roots and shoots. Sources showed no differences concerning TLC plates. TLC analysis did not detect β-ecdysone in the in vitro cultured plants.

Pfaffia glomerata (Spreng.) Pedersen, conhecida como ginseng brasileiro, é muito utilizada pela medicina popular devido suas propriedades fitoterápicas. Este trabalho teve como objetivo avaliar o efeito de concentrações de BAP e de TDZ na propagação in vitro de dois acessos (BRA e JB-UFSM) desta espécie. Segmentos nodais, provenientes de plantas assépticas, foram cultivados em meio Murashige e Skoog (MS) suplementado com 0, 1 e 5 μM de BAP ou TDZ. Após 30 dias, as plantas foram transferidas para meio MS não suplementado com citocinina. Observou-se que a organogênese in vitro de P. glomerata é genótipo-dependente. O acesso BRA mostrou um maior potencial para a propagação in vitro em relação ao JB-UFSM. O cultivo dos segmentos nodais do acesso BRA em meio com 1 μM de TDZ, seguido pelo subcultivo dos brotos para meio isento de citocininas mostrou ser um método viável para a propagação in vitro devido à alta taxa de multiplicação e o bom desenvolvimento das plantas. No acesso JB-UFSM, o cultivo dos segmentos nodais em meio não acrescido de citocinina mostrou ser o método mais adequado para a produção de um grande número de plantas com alta qualidade.The plant Pfaffia glomerata (Spreng.) Pedersen, known as Brazilian ginseng, is extensively used in folk medicine due to its phytotherapic characteristics. This work aimed to evaluate the effect of BAP and TDZ concentrations on the in vitro propagation of two sources (BRA and JB-UFSM) of this species. Nodal segments originated from aseptically grown plants were cultured on Murashige and Skoog (MS) medium supplemented with 0, 1 and 5 μM BAP or TDZ. After 30 days, the plants were transferred to MS medium without cytokinin. The in vitro organogenesis of P. glomerata is genotype-dependent. The source BRA had greater potential for in vitro propagation than JB-UFSM. The culture of BRA nodal segments on medium with 1 μM TDZ, followed by subcultivation of sprouts on cytokinin-free medium, showed to be a viable method for in vitro propagation due to high multiplication rate and good plant development. In the source JB-UFSM, the culture of nodal segments on medium without cytokinin was the best method to obtain a large number of high-quality plants.

Este trabalho teve por objetivo caracterizar as exigências nutricionais de mudas de ginseng brasileiro (Pfaffia glomerata (Spreng.) Pedersen) em Argissolo Vermelho distrófico arênico pela técnica do nutriente faltante. Plantas de P. glomerata produzidas in vitro e aclimatizadas ex vitro foram usadas como material inicial para o experimento em casa de vegetação. Aos 10 dias após o transplante para o solo, através de soluções nutritivas, foram realizadas as adubações correspondentes a nove tratamentos: controle (sem adubação), adubação completa (com macro e micronutrientes) e as omissões individuais de N, P, K, Ca, Mg, S e dos micronutrientes (Zn, B, Cu, Fe, Mn e Mo). Aos 60 dias de cultivo, avaliaram-se os parâmetros de crescimento das plantas. O número de folhas por planta foi o parâmetro de crescimento mais afetado pela ausência dos nutrientes N, K, S e P na adubação. O efeito da omissão dos nutrientes na adubação foi diferenciado quanto à partição de biomassa entre os órgãos da planta. A biomassa acumulada pela parte aérea obedeceu à seguinte ordem decrescente, em relação à omissão do nutriente: Mg @ micronutrientes > Ca @ adubação completa > P > S > K > N @ controle. Já a biomassa radicular obedeceu à seguinte ordem decrescente: Mg @ adubação completa ³ P ³ micronutrientes ³ S ³ Ca > N > K @ controle. Plantas jovens de P. glomerata cultivadas em Argissolo Vermelho distrófico arênico apresentaram grande redução no crescimento pela ausência de N, K, S e P na adubação. As omissões de Mg e dos micronutrientes na adubação não foram limitantes para o crescimento das plantas jovens.This study aimed to characterize the nutritional demands of the young plants of Brazilian ginseng (Pfaffia glomerata (Spreng.) Pedersen) in a Paleudalf soil by means of the missing nutrient technique. Plantlets of P. glomerata produced in vitro and acclimatized ex vitro were used as start matter for the glasshouse experiment. Ten days after plant transfer to soil, fertilizations (by means of nutritive solutions) were done corresponding to nine treatments: control (without fertilization), complete fertilization (with macro and micronutrients), and with the omissions of N, P, K, Ca, Mg, S and micronutrients (Zn, B, Cu, Fe, Mn and Mo). At 60 days of cultivation, several growth parameters were evaluated. The number of leaves per plant was the growth parameter most affected by omission of N, K, S and P in the fertilization. The effect of nutrient omission in the fertilization on biomass partitioning between plant organs were differentiated. Accumulated shoot biomass obeied the following decreasing order, in relation with the nutrient omission: Mg @ micronutrients > Ca @ complete fertilization > P > S > K > N @ control. While the root biomass obeied the following decreasing order: Mg @ complete fertilization ³ P ³ micronutrients ³ S ³ Ca > N > K @ control. Young plants of P. glomerata grow in a Paleudalf soil showed greater reduction in growth by omission of N, K, S and P in the fertilization. The omission of Mg and micronutrients in the fertilization were not limiting to the growth of young plants.

O presente trabalho objetivou avaliar o crescimento de ginseng brasileiro (Pfaffia glomerata (Spreng.) Pedersen) cultivado in vitro sob dois níveis de nitrogênio e sacarose, durante seis subculturas sucessivas e aclimatização. Como fonte de explantes, utilizou-se segmentos nodais de plantas previamente estabelecidas in vitro. Foram testados dois tratamentos: 100% da concentração padrão de N do meio MS + 30g L-1 sacarose; e 50% da concentração padrão de N do meio MS + 45g L-1 sacarose. Durante as seis subculturas sucessivas, os explantes de plantas cultivadas por 30 dias em ambos os tratamentos foram transferidos, respectivamente, para a subcultura seguinte nos mesmos tratamentos. Ao final da 6ª subcultura, as plântulas foram transferidas para substrato Plantmax® Hortaliças e submetidas à aclimatização em sala de crescimento. Durante as subculturas, observou-se flutuações nas respostas de crescimento das plântulas às duas condições nutricionais testadas. A altura da maior brotação, a média da altura das brotações e o total de segmentos nodais por plântula, respectivamente em quatro, quatro e três subculturas, foram maiores em 100% da concentração padrão de N do meio MS + 30g L-1 sacarose do que em 50% da concentração padrão de N do meio MS + 45g L-1 sacarose. Por outro lado, as matérias secas de raízes, da parte aérea e do total das plântulas, respectivamente em duas, duas e três subculturas, foram maiores em 100% da concentração padrão de N do meio MS + 30g L-1 sacarose. Concluiu-se que o aumento da concentração de sacarose associado à redução da concentração do N estimula a produção de biomassa, enquanto o meio MS padrão estimula o crescimento em altura e número de segmentos nodais. As diferenças quanto à produção de biomassa observadas no cultivo in vitro não se refletiram no processo de aclimatização ex vitro, onde o meio MS padrão proporcionou maior crescimento em altura e número de folhas por planta.The present research was aimed at evaluating the growth of brazilian ginseng (Pfaffia glomerata (Spreng.) Pedersen) in vitro cultured plantlets under two levels of nitrogen and sucrose, during six successive subcultures and acclimatization. Two nutritional treatments were tested: nitrogen at 100% strength of the MS medium + 30g L-1 sucrose; and nitrogen at 50% strength + 45g L-1 sucrose. During six successive subcultures, the explants of 30-day-old plantlets grown under both treatments were transferred, respectively to the next subculture on the same treatment conditions. At 6th subculture, plantlets were transferred to substrate Plantmax® Hortaliças and submitted to acclimatization in a growth chamber. Throughout the subcultures, fluctuations in growth response due to nutritional conditions tested were observed. The height of the higher sprout, the average height of sprouts, and the total number of nodal segments per plantlet, respectively for four, four and three subcultures were greater in 100% strength of the MS medium + 30g L-1 sucrose than in 50% strength + 45g L-1 sucrose. On the other hand, dry matter of roots, of shoots and of the whole plant, respectively for two, two and three subcultures, were greater in 50% strength + 45g L-1 sucrose. In conclusion, the increase of sucrose associated with reduction of N level in the culture medium enhanced the biomass production, while the MS at full strength enhanced the growth based in sprout length and number of nodal segments. These differences concerning the biomass production observed during in vitro culture did not persisted in the acclimatization process ex vitro, where MS at full strength give rise greater growth in sprout length and number of leaves per plant.

Pfaffia glomerata (Spreng) Pedersen is a medicinal plant largely used as an adaptogenic herb. The species is naturally found in Africa and Americas and is the subject of pharmaceutical and commercial interest, mainly due to the accumulation of β-ecdysone in its roots. Brazil is considered the greatest supplier of P. glomerata in the world. Due to the morphological similarity of its roots to those of Panax ginseng (Korean ginseng), the species became known as the Brazilian ginseng. In this study, we performed the cytogenotoxic evaluation of P. glomerata root extract in rodent bone marrow micronucleus and Allium cepa assays. Taking into consideration the therapeutic dosage of P. glomerata , three methanolic extract dilutions were used in the Micronucleus assay. Each animal was treated with two consecutive doses with a 24-h interval. To assess the cytogenotoxicity of the extract, 2000 polychromatic erythrocytes per animal were analyzed. For the Allium cepa assay, onion seeds were exposed during 24 h to six methanolic extract dilutions. Cell cycle and chromosomal alterations were quantified. The results of the Micronucleus assay showed genotoxic effects for all treatments investigated, considering the increase in the number of micronucleated cells. In the A. cepa assay the methanolic extract induced alterations in the cell cycle. Aneugenic and clastogenic effects were observed. The results indicate cytogenotoxic activity of P. glomerata extract in animal and plant cells. Thus, the ingestion of P. glomerata for medicinal purposes should be done carefully. The results also lead to the possibility of investigating the potential of the species in the research of prospective compounds for cancer treatment and for isolation of biologically active substances. [ABSTRACT FROM AUTHOR]

The present study aimed to verify the effects of induced polyploidization on Pfaffia glomerata regarding its 20-hydroxyecdysone (20E) production both in vitro and under greenhouse conditions, its in vitro photoautotrophic potential, and its ex vitro biomass accumulation and photosynthetic performance. Synthetic polyploidization efficiently produced individuals with increased in vitro photoautotrophic potential and ex vitro biomass accumulation, although photosynthetic rates per leaf area did not vary between diploids and tetraploids. Among the five tetraploids tested (P28, P60, P68, P74, and P75), P28 showed significantly increased biomass both in vitro and ex vitro when compared with diploid plants, whereas the other tetraploids did not differ significantly from the diploids in terms of biomass accumulation. Although photosynthetic rates per unit leaf area remained constant among all the plants tested, P28 showed a significantly greater total leaf area, which may have resulted in an increase in net photosynthesis on a whole-plant basis. Under greenhouse conditions, the 20E content in the tetraploid P28 was 31% higher than that in diploid plants, and the final 20E mass per plant produced by P28 ex vitro was approximately twice that produced by diploid plants. Accumulation of 20E in vitro did not follow the same pattern observed among the plants ex vitro; instead, greater accumulation was observed in diploid plants. The induction of polyploidy in P. glomerata appears to be a promising strategy for producing plants with higher biomass accumulation and 20E production ex vitro, in addition to its higher in vitro photoautotrophic potential. [ABSTRACT FROM AUTHOR]

The mineral nutritional homeostasis in response to different concentrations of Hg (0, 25 and 50µM) was evaluated in Pfaffia glomerata plant. The exposure to the highest level of Hg (50µM) caused a decreasing in shoot and root fresh weights of 15.5% and 20%, respectively. Both shoot and root Hg concentrations increased linearly with increasing external Hg concentrations. Ca concentration decreased in shoot only at 50µM Hg, whereas shoot K and Mg concentrations decreased at both 25 and 50µM Hg, when compared to the control. A significant decrease in Cu, Zn, Fe and Mn concentrations in plants exposed to Hg was observed, but most Zn, Mn, and Cu in the roots. On the other hand, P concentration increased in both root and shoot of plants exposed at 25 and 5µM Hg, whereas Na concentration increased only in the root at 25 and 50µM Hg exposure. In general, tissue nutrient concentrations in P. glomerata plantlets exposed to Hg were significantly decreased, which indicates that the Hg may cause alteration on the mineral nutritional homeostasis of this species. [ABSTRACT FROM AUTHOR]

Ethnaopharmacologial relevance In South America, the β-ecdysone ecdysteroid has been found in species of the genus Pfaffia Mart. Due to the similar morphology of its roots to the Panax ginseng C. A. Mey. (Korean ginseng), some species of this genus has been known as Brazilian ginseng and have been used as tonic and aphrodisiac, as well as for the treatment of diabetes and rheumatism. Aim of the study Here we report a cytogenotoxic evaluation of β-ecdysone (a natural ecdysteroid found in plants) in Rodent Bone Marrow Micronuclei and Allium cepa Assays. Materials and methods Three β-ecdysone (pure) concentrations (based in human therapeutic dosage) were used in the Micronucleus Assay. The animals were treated during two consecutive days. Micronucleated cells were counted in 2000 polychromatic erythrocytes per animal. For A. cepa L. Assay, one β-ecdysone concentration was analyzed. The onions bulbs were exposed for 24 h. Results The Micronucleus Assay showed genotoxic effects for all treatments, expressed by an increase of micronucleated cells. In A. cepa L. Assay, cell abnormalities associated to the malfunction/non-formation of mitotic spindle (aneugenic effect) and chromosomal bridges (clastogenic effect) were observed. Conclusions The results indicate a cytogenotoxic activity of β-ecdysone. Therefore, the popular use of Pfaffia and others species containing β-ecdysone should be considered with caution. [ABSTRACT FROM AUTHOR]

[Objective] This study aimed to explore the pharmacognostical features of Brazilian ginseng (Pfaffia paniculata), and to provide a theoretical basis for its development and utilization. [Method] Morphological traits of Brazilian ginseng root were observed with traditional methods. Then, the root, stem and leaf of Brazilian ginseng were sliced, and observed under a microscope. The physical and chemical properties of root extract were detected. [Result] Brazilian ginseng root is elongated and cylindrical, with obvious longitudinal ridges and sparse root marks, some irregularly shaped lenticels on the surface. The cork layer of crosssection has five or more rows of cork cells. The abnormal vascular bundles are arranged in two to three rings. There are two to three central vascular bundles. The wood fibers in powder are long, with one bifurcate end. The vessels have bordered pits. The sand crystals are scattered or in the parenchyma surrounding the fibers. The mucilage cells are oval and widely exist. Physical and chemical experiments showed that the root of Brazilian ginseng contains sterols or triterpenes and saponins. [Conclusion] The results will provide a theoretical basis for the pharmacological identification of Brazilian ginseng. [ABSTRACT FROM AUTHOR]

Aluminum (Al) toxicity is a limiting factor for crop production in acid soils, which cover approximately 60% of the Brazilian territory. This study aimed to evaluate the effects of Al on growth and tissue Al concentration of two Pfaffia glomerata accessions (BRA and JB/UFSM). Plantlets were grown in a hydroponic system with five Al concentrations (0, 50, 100, 150 and 200mg L-1) for 7 days. Most of the evaluated parameters presented significant interaction between both P. glomerata accessions and Al levels in nutrient solution and, in general, Al treatments negatively affected plant growth, especially roots. Moreover, BRA accession showed higher Al accumulation in its tissues than JB/UFSM and, consequently in BRA accession the growth was impaired substantially. Furthermore, the results suggest that, between P. glomerata accessions studied, BRA is less appropriated for medicinal uses when grown in soils with high Al levels, due to the higher accumulation of tissue Al content. [ABSTRACT FROM AUTHOR]

This study aimed to investigate whether the in vitro photoautotrophic potential and 20 hydroxyecdysone (20E) production of six Pfaffia glomerata accessions could be correlated to photosynthetic performance and biomass accumulation under ex vitro conditions in a greenhouse environment. Our results showed that P. glomerata accessions displayed varying photosynthetic rates that may have resulted in differential biomass accumulation both in vitro and ex vitro. Accessions A4, A13 and A43 showed the highest dry weight under in vitro photoautotrophic conditions, coinciding with the higher photosynthetic rates of the same accessions in greenhouse. They also showed the highest 20E mass per plant ex vitro. Production of 20E in vitro also varied among accessions, with the highest total mass achieved by accessions A4 and A43. In addition to having the potential for optimizing propagation by reducing the duration of the in vitro culture stage and lessening death rate during acclimatization, the use of these genotypes with higher in vitro photoautotrophic potential can guarantee highly productive plants ex vitro because the biomass accumulation pattern observed in vitro matched that under greenhouse conditions. [ABSTRACT FROM AUTHOR]

Abstract: Hebanthe eriantha (Poir.) Pedersen (Amaranthaceae), which is known as Brazilian ginseng is widely used in folk medicine as an aphrodisiac and antidiabetic tonic. The anti-tumor activity, attributed to the pfaffic acid present in roots of H. eriantha, is responsible for the great interest in the commercialization of this species. In Brazil, the species H. eriantha is mainly used in commercial preparations, although other plants of the genus Pfaffia and Hebanthe have been marketed as Pfaffia paniculata or Brazilian ginseng. The pfaffic acid present in the roots is mainly conjugated with sugars (pfaffosides) and can be used as an active marker of H. eriantha, which helps to differentiate this species from others marketed as Brazilian ginseng. The main objective of this study was to develop and validate a liquid chromatographic method to quantify pfaffic acid in the roots of H. eriantha. The extraction and hydrolysis conditions were optimized using an univariate and experimental design, respectively, and the quantification of pfaffic acid by high performance liquid chromatography with diode-array detection (HPLC-DAD) was validated. This method was used to evaluate the pfaffic acid content in 30 different genotypes of the species from a germplasm collection. The content of pfaffic acid ranged from 0.97 to 4.29% (w/w) on a dry weight basis. [Copyright &y& Elsevier]

The possible role of zinc (Zn) to reverse the oxidative stress caused by mercury (Hg) was investigated in Pfaffia glomerata plantlets. Thirty-day-old acclimatized plantlets of P. glomerata were exposed to four treatments: control, 50 μM Zn, 50 μM Hg and 50 μM Zn + 50 μM Hg for 9 days. In Zn + Hg treatment, shoot and root Hg concentrations were 59 and 24% smaller than that plants exposed to 50 μM Hg added alone. An increase in the Zn concentration in the shoot of plants exposed to Zn + Hg occurred, although in the roots Zn concentration was not altered, when compared to the control. Fresh and dry weights, as well as the activity of δ-aminolevulinic acid dehydratase (δ-ALA-D) in Hg-treated plants were significantly reduced. Percentage survival, fresh and dry weights and δ-ALA-D activity of plants treated by 50 μM Zn + 50 μM Hg were greater than of that treated by Hg alone. Moreover, Zn treatment reduced the lipid peroxidation caused by Hg, being this effect related to increased root superoxide dismutase activity, and shoot catalase and ascorbate peroxidase activities. In conclusion, the presence of Zn in the substrate caused a significant reduction in the oxidative stress induced by Hg. [ABSTRACT FROM AUTHOR]

Pfaffia glomerata is a medicinal plant widely distributed in Brazil, which is considered the world's greatest supplier of P. glomerata roots. Among active ingredients contained in this plant, the steroid β-ecydisone (20E) is the most important compound extracted from roots. This steroid presents therapeutic properties for the treatment of diabetes and haemorrhoids, besides having bioenergy, tonic and aphrodisiac effects. The root-knot nematode Meloidogyne spp. is a major limiting factor in root production. Recent studies showed resistance of accessions of P. glomerata to Meloidogyne incognita. The aims of this work were: i) to correlate the concentration of 20E with resistance and susceptibility of P. glomerata accessions to M. incognita in inoculated and non-inoculated plants; ii) to study the effect of the parasitism of M. incognita on the concentration of the steroid 20E in the roots; and iii) to clarify resistance mechanisms by comparing the response of a highly resistant UFV with a highly susceptible accession (Farm) to nematode infection. The concentration of 20E in the healthy susceptible Farm accession was significantly higher than in the healthy resistant UFV accession, showing that the resistance mechanism was not related to 20E concentrations. Plants of the Farm accession infected with M. incognita showed higher levels of 20E than the non-infected control. A positive and significant Pearson correlation coefficient was observed between 20E concentrations and gall indexes. Resistance of UFV to the root-knot nematode M. incognita was associated with unidentified factors that limited nematode penetration or emigration of second-stage juveniles and with post-penetration responses, including the hypersensitive response. Giant cells were sometimes found in the resistant cultivar, but displayed a highly vacuolated and degraded cytoplasm with thinner cell walls than those induced in the susceptible accession. Microscope observations under UV light showed a strong autofluorescence, suggesting that phenolic compounds may be involved in ginseng UFV resistance. [ABSTRACT FROM AUTHOR]

Abstract: Brazilian Ginseng extracts of two species, Pfaffia paniculata and Pfaffia glomerata, were obtained by supercritical fluid extraction (SFE) with CO2 and by low-pressure solvent extraction (LPSE) with methanol, hexane and ethanol. The SFE assays were conducted at pressures of 100, 200 and 300bar, and temperatures of 30 and 50°C. The qualitative chemical compositions of the extracts were determined by thin layer chromatography (TLC). One of the active principles of interest from P. glomerata extract, β-ecdysone, was identified and quantified by HPLC. The antioxidant activities of Brazilian Ginseng extracts were determined by the coupled reaction of linolenic acid and β-carotene. For P. paniculata, the highest SFE yield was obtained at 200bar/50°C (0.22%, dry basis—d.b.), while the best extraction condition for P. glomerata was obtained at 200bar/30°C (0.18%, d.b.). The higher extract yields obtained by LPSE were 2.0% and 5.8% (w/w, d.b.) for P. paniculata and P. glomerata, respectively, both obtained with methanol as extraction solvent. From the overall extraction curve of P. glomerata, it was possible to obtain the kinetic parameters of extraction; the duration of the CER (constant extraction rate) period was determined as 134min. The TLC plates showed the possible presence of flavonoids in the ethanolic extract for both Pfaffia species. The antioxidant activity analysis detected that LPSE extracts had higher activity than SFE extracts. [Copyright &y& Elsevier]

The aim of this study was to investigate both a mass in vitro propagation system and the β-ecdysone content in roots and aerial parts of Pfaffia glomerata and Pfaffia tuberosa. Nodal segments of two genotypes (BRA and JB-UFSM) of P. glomerata, originated from aseptically grown plants, were cultivated on hormone-free Murashige and Skoog medium. For the proliferation of P. tuberosa shoots, nodal segments, originated from aseptically grown plants, were either cultivated on hormone-free Murashige and Skoog (MS) medium or were supplemented with 1.0 µM thidiazuron (TDZ); the elongation and rooting of these plants were carried out on MS medium without TDZ. Plantlets of both species were acclimatized and transferred to field conditions. The β-ecdysone content in the plants was determined by high performance liquid chromatography. The BRA genotype showed a greater in vitro proliferation rate and β-ecdysone content than that of the JB-UFSM genotype. The culture of nodal segments of P. tuberosa on medium with 1.0 µM TDZ with subsequent subcultivation of shoots on hormone-free medium was shown to be a suitable method for micropropagation due to the high multiplication rate and good plant development. Both species showed good adaptation to ex vitro conditions. The β-ecdysone content in micropropagated P. tuberosa was similar to that found in field-grown plants. For both species, the aerial parts accumulated higher β-ecdysone content than roots. These results reveal that micropropagation is a successful, alternative method for rapid plant multiplication of both species of Brazilian ginseng. Furthermore, this study demonstrates that these two species have a potential for cultivation that is associated with high β-ecdysone production. [ABSTRACT FROM AUTHOR]

Abstract: Pfaffia paniculata (Brazilian ginseng) roots and/or its extracts have shown anti-neoplastic, chemopreventive, and anti-angiogenic properties. The aim of this work was to investigate the chemopreventive mechanisms of this root in mice submitted to the infant model of hepatocarcinogenesis, evaluating the effects on cellular proliferation, apoptosis, and intercellular communication. Fifteen-day-old BALB/c male mice were given, i.p., 10μg/g of the carcinogen N-nitrosodiethylamine (DEN). Animals were separated into three groups at weaning and were given different concentrations of powdered P. paniculata root (0%, 2%, or 10%) added to commercial food for 27 weeks. Control group (CT) was not exposed to the carcinogen and was given ration without the root. After euthanasia, the animals’ liver and body weight were measured. Liver fragments were sampled to study intercellular communication, molecular biology, and histopathological analysis. Cellular proliferation was evaluated by immunohistochemistry for PCNA, apoptosis was evaluated by apoptotic bodies count and alkaline comet technique, and intercellular communication by diffusion of lucifer yellow dye, immunofluorescence, western blot and real-time PCR for connexins 26 and 32. Chronic treatment with powdered P. paniculata root reduced cellular proliferation and increased apoptosis in the 2% group. Animals in the 10% group had an increase in apoptosis with chronic inflammatory process. Intercellular communication showed no alterations in any of the groups analyzed. These results indicate that chemopreventive effects of P. paniculata are related to the control of cellular proliferation and apoptosis, but not to cell communication and/or connexin expression, and are directly influenced by the root concentration. [Copyright &y& Elsevier]

Abstract: We have previously reported a reduction in the accumulation of ascitic fluid in Ehrlich tumor-bearing mice following treatment with the powdered roots of Pfaffia paniculata. The aim of this study was to investigate which extracts from these roots presented antineoplastic properties. Thus, the effects of the ethanolic extract, butanolic residue, or aqueous residue from Pfaffia paniculata on animal survival and tumor growth in mice bearing this tumor were studied. Butanolic residue-treated mice survived longer than untreated mice. This result points to an antineoplastic effect exerted by the butanolic fraction from the roots of P. paniculata on this tumor model. [Copyright &y& Elsevier]

Objective: To evaluate the free radical scavenging and cytotoxic activities of the butanolic (BuOH) extract, methanolic (MeOH) extract and 20-hydroxyecdysone extracted from the roots of Pfaffia glomerata. Materials and methods: Pfaffia glomerata roots were collected, powdered and extracted with methanol by maceration at room temperature. The extract was concentrated under vacuum, yielding a residue, followed by a butanol extraction. The 20-hydroxyecdysone (EC) was obtained by chromatographic separation of the BuOH fraction. An amount of 10 mg of each dry extract and EC was dissolved in 0.1% dimethyl sulphoxide-phosphate-buffered-saline solution (DMSO-PBS) and screened for their capabilities on scavenging thiobarbiturate reactive substances (TBARS). The antioxidant activity of each extract was determined in vitro by measuring malonyldialdehyde (MDA) and 4-hydroxynonenal (4-HNE) in erythrocyte ghosts treated with ferric-ascorbate. The investigation has also included the cytotoxicity measurement by Trypan blue exclusion test and tetrazolium reduction assay in mice peritoneal macrophages. Results: The free radical scavenging activity of EC was higher than that present in the BuOH fraction. The MeOH extract showed a remarkable pro-oxidant activity. The EC-free radical reaction-inhibition was almost twice of that of the control α-Tocopherol (αT). The Trypan blue exclusion assay confirmed toxicity of the MeOH extract, whose lethality surpassed 80% of the treated macrophages after 1 h of 0.01 mg exposure per 106 cells. Conclusions: The present study shows the antioxidant effect of the Brazilian Ginseng. The scavenging effect was evidenced for EC as well the BuOH fraction. The MeOH extract showed cytotoxicity on mice peritoneal macrophages. Such toxicity is probably due to ginsenosides present in this latter fraction and warrants further toxicological evaluation of the Brazilian Ginseng roots. [ABSTRACT FROM AUTHOR]

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. 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. 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. 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). Image 1 [ABSTRACT FROM AUTHOR]

Zinc (Zn) is a micronutrient, but its excessive concentration can impair plant growth and development. Fertilizers, liming materials, pesticides and fungicides containing Zn have contributed to increase its concentration in agricultural soils. The aim of the present study is to evaluate the effect of Zn excess on the non-enzymatic (anthocyanin and β-ecdysone) and enzymatic (superoxide dismutase-SOD and guaiacol peroxidase-GPX) antioxidant system of two P. glomerata accessions (JB and GD) grown in hydroponic system and soil, under short- and long-term exposure times. Three Zn levels (2, 100 and 200 μM) and two short-term exposure times (7 and 14 d) were tested in the hydroponic experiment. Three Zn levels (2, 100 and 200 mg kg−1) and two long-term exposure times (34 and 74 d) were tested in the soil experiment. The effects of Zn excess on P. glomerata accessions depended on the growth system and exposure time. Zinc excess in both tested growth systems resulted in significant change in the tissue oxidative process (MDA concentration) in both accessions, as well as broadened the antioxidant system response, which was based on antioxidant enzymes (SOD and GPX) and secondary metabolites (anthocyanins and β-ecdysone). The highest anthocyanin concentration was observed in accession JB, which was grown in hydroponics, but tissue anthocyanin concentration increased in both accessions, regardless of growth medium and exposure time. The β-ecdysone concentration in the roots increased in both accessions, but accession GD was more responsive to Zn excess. There was significant physiological variation in P.glomerata accessions in response to Zn excess. • Shoots had higher Zn concentration than roots, regardless of cultivation system. • Shoot anthocyanin concentration increased regardless of exposure time to Zn excess. • Root β-ecdysone concentration increased as Zn concentration increased. • There was physiological variability in P.glomerata accessions to due Zn excess. • P.glomerata has moderate tolerance to Zn excess. [ABSTRACT FROM AUTHOR]

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. Unlabelled Image • A 1R:1B ratio increases content and production of 20E by 30–40% in P. glomerata. • A 1R:1B ratio increases biomass, anthocyanins, and lignification of vascular tissue. • A 1R:3B ratio improves photosynthesis, electron transport, and antioxidant activity. • Primary metabolism responds differently to spectral quality in Ac22 and Ac43 plants. [ABSTRACT FROM AUTHOR]