Bianca Waléria Bertoni

Antihepatotoxic agent in micropropagated plantlets of Eclipta alba

A protocol for in vitro micropropagation of Eclipta alba (L.) Hassk from nodal segment explants has been established. The maximum number of shoots was obtained after 60 days of culture in Murashige & Skoog (MS) medium supplemented with 4.4 μM benzyladenine. Multiple rooting was achieved using MS medium with 2.4 μM 2-isopentyladenine. Wedelolactone was present in shoots cultured in media containing cytokinins.

Micropropagation of Pothomorphe umbellata via direct organogenesis from leaf explants

The establishment of a micropropagation protocol for Pothomorphe umbellata was carried out using leaf segments cultured on 1/4 strength Murashige and Skoog medium supplemented with 0.5 mg l-1 6-benzyladenine, 0.1 mg l-1 gibberelic acid added with 10 g l-1 sucrose. Rooting was achieved using MS medium devoid of growth regulators. An anatomical study confirmed shoot regeneration via direct organogenesis.

Genetic diversity in natural populations of Jacaranda decurrens Cham. determined using RAPD and AFLP markers

Jacaranda decurrens (Bignoniaceae) is an endemic species of the Cerrado with validated antitumoral activity. The genetic diversity of six populations of J. decurrens located in the State of São Paulo was determined in this study by using molecular markers for randomly amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP). Following optimization of the amplification reaction, 10 selected primers generated 78 reproducible RAPD fragments that were mostly (69.2%) polymorphic. Two hundred and five reproducible AFLP fragments were generated by using four selected primer combinations; 46.3% of these fragments were polymorphic, indicating a considerable level of genetic diversity. Analysis of molecular variance (AMOVA) using these two groups of markers indicated that variability was strongly structured amongst populations. The unweighted pair group method with arithmatic mean (UPGMA) and Pearsons correlation coefficient (RAPD -0.16, p = 0.2082; AFLP 0.37, p = 0.1006) between genetic matrices and geographic distances suggested that the population structure followed an island model in which a single population of infinite size gave rise to the current populations of J. decurrens, independently of their spatial position. The results of this study indicate that RAPD and AFLP markers were similarly efficient in measuring the genetic variability amongst natural populations of J. decurrens. These data may be useful for developing strategies for the preservation of this medicinal species in the Cerrado.

A standardized methanol extract of Eclipta prostrata (L.) L. (Asteraceae) reduces bronchial hyperresponsiveness and production of Th2 cytokines in a murine model of asthma

ETHNOPHARMACOLOGICAL RELEVANCE Eclipta prostrata (L.) L. (Asteraceae) has been used in Brazilian traditional medicine to treat asthma and other respiratory illnesses. AIMS OF THE STUDY To investigate the effects of different doses of a standardized extract of E. prostrata using a murine model of allergen induced asthma. MATERIALS AND METHODS Balb/c mice were sensitized twice with ovalbumin (OVA) administered intraperitoneally and challenged over four alternate days with nasal instillations of OVA solution. The standardized methanol extract of E. prostrata was administered in doses of 100, 250 and 500mgkg-1 concomitantly with nasal instillation over seven consecutive days. Control animals were treated with dexamethasone or saline solution. Bronchial hyperresponsiveness, production of Th1 and Th2 cytokines, allergen sensitization, airway and lung inflammation, mucous secretion and airway remodeling were assessed. RESULTS The concentrations of chemical markers in the standardized methanol extract were 0.02% oroboside, 1.69% demethylwedelolactone and 1.71% wedelolactone. Treatment with 250mgkg-1 of extract, which provided 0.745, 4.22 and 4.30mgkg-1day-1 of oroboside, demethylwedelolactone and wedelolactone, respectively, significantly reduced (P<0.05) respiratory resistance and elastance. Such effects were comparable with those produced by dexamethasone. The total number of inflammatory cells and eosinophils in the bronchoalveolar lavage and the concentrations of interleukin (IL)-4, IL-5 and IL-13 in lung homogenate were significantly reduced (P<0.05) by the methanol extract of E. prostrata. CONCLUSION The results presented herein demonstrate for the first time the anti-inflammatory activity of E. prostrata in a murine model of asthma, thereby supporting the ethnopharmacological uses of the plant.

An ent-kaurane-type diterpene in Croton antisyphiliticus mart.

Croton antisyphiliticus is a medicinal plant widely used in the treatment of microbial infections, especially those affecting the genital tract. Crude extract, fractions and pure compound isolated from roots of this species were investigated to validate their antimicrobial activity against Escherichia coli and Staphylococcus aureus. The compound ent-kaur-16-en-18-oic acid was isolated as a major component (0.7% of crude extract), and its MIC value determined against S. aureus (ATCC 6538) was 250 μg/mL. This is the first phytochemical work on the species monitored with antimicrobial assay.

Determination of lead content in medicinal plants by pre-concentration flow injection analysis-flame atomic absorption spectrometry.

INTRODUCTION Although medicinal plants are widely used throughout the world, few studies have been carried out concerning the levels of heavy metal contaminants present. Such metals are highly toxic to living organisms even in low concentrations owing to their cumulative effect. The present paper describes the the development of a pre-concentration flow injection analysis-flame atomic absorption spectrometric system to determine the lead content in medicinal plants at the ppb level. OBJECTIVE To develop a pre-concentration flow injection analysis-flame atomic absorption spectrometric system to determine the lead content in medicinal plants at the ppb level. METHODOLOGY A pre-concentration flow system was coupled to a flame atomic absorption spectrometer. The plant samples were analysed after nitroperchloric digestion. The proposed system was optimised by evaluating the following parameters: nature, concentration and volume of the eluent solution, elution flow rate, elution efficiency, pre-concentration flow rate and pre-concentration time. RESULTS The proposed system exhibited good performance with high precision and repeatability (RSD < or = 2.36%), excellent linearity (r = 0.9999), low sample consumption (10.5 mL per determination) and an analytical throughput of 55 samples/h. Lead concentrations ranged from 3.37 + or - 0.25 to 7.03 + or - 0.51 microg/g in dry material. This concentration interval is greater than that previously published in the literature. CONCLUSION The inclusion of a pre-concentration column in the flow manifold improved the sensitivity of the spectrometer. Thus, it was possible to determine the analyte at the ng/mL level in sample solutions of medicinal plants. This is a very important accomplishment, especially when the cumulative effect of heavy metals in living organisms is considered.

Micropropagação de Dioscorea multiflora Grised

Dioscorea multiflora is a plant native to southern Brazil that produces diosgenin as a major secondary metabolite, a substance which is used by the pharmaceutical industry for the production of cortisone and substances with contraceptive action. The objective of this work was to optimize the micropropagation protocol of D. multiflora, for the production of seedlings on a commercial scale. Nodal segments subcultured in solid MS medium were transferred for multiplication to MS medium supplemented with BAP (0.01, 0.1, 0.5, 1.0 and 3.0 mg L-1) and MS medium supplemented with 0.1 mg L-1 or 0.5 mg L-1 BAP plus different concentrations of sucrose (2, 4, 6, 8 and 10%). For rooting, the shoots were cultured on MS medium supplemented with IBA (0.1, 0.5, 1.0 and 3.0 mg L-1) and MS medium supplemented with NAA (0.1, 0.5, 1.0 and 3.0 mg L-1). A completely randomized design was used with treatment consisting of 3 replicates with 10 buckets per plot. Seedlings with and without roots were acclimatized in a greenhouse. The best results of multiplication and rooting were obtained in MS medium + 0.1 mg L-1 BAP (80%) and in MS medium + 1.0 mg L-1 IBA (42.6%), respectively. There was no difference in the survival percentage of seedlings in vitro and during ex vitro acclimatization (75%). The micropropagation protocol for production of D. multiflora is effective and can be used for commercial production.

Genetic variability in natural populations of Zeyheria montana mart. from the Brazilian Cerrado

Zeyheria montana, an endemic species of the Bignoniaceae family from the Brazilian Cerrados known for its anti-cancer properties, is widely used as imuno stimulant in the popular medicine and its therapeutic activity must be validated by scientific data. The objective of this work was to evaluate the genetic variability of eight plant populations collected within the state of Sao Paulo, Brazil, via Random Amplification of Polymorphic DNA (RAPD) used as molecular markers. After an optimized protocol for the amplification reaction, nine selected primers generated 105 reproducible bands, indicating up to 60% polymorphism. Analysis of molecular variance (AMOVA) revealed higher genetic variation within populations (84.03%) than among populations (15.97%). The variation values estimated by φ ST (0.160) indicated moderate to high inter population structuration. Levels of similarity inter plants with genetic and geographical distances, estimated by the unweighted pair-group method analysis (UPGMA) clustering and non-metric multidimensional scaling (NMDS) ordination methods and by the Mantel test (-0.2345 p = 0.118) denoted that the structure found follows the island model, which assumes that a single population of infinite size may have initiated the existing populations of Zeyheria montana, with no spatial position correlation. Based on the obtained data, a germplasm bank from individuals representing the species variability was established. Furthermore the information here reported can be of importance to develop strategies for the conservation of Z. montana.

Genetic characterization of fig tree mutants with molecular markers

The fig (Ficus carica L.) is a fruit tree of great world importance and, therefore, the genetic improvement becomes an important field of research for better crops, being necessary to gather information on this species, mainly regarding its genetic variability so that appropriate propagation projects and management are made. The improvement programs of fig trees using conventional procedures in order to obtain new cultivars are rare in many countries, such as Brazil, especially due to the little genetic variability and to the difficulties in obtaining plants from gamete fusion once the wasp Blastophaga psenes, responsible for the natural pollinating, is not found in Brazil. In this way, the mutagenic genetic improvement becomes a solution of it. For this reason, in an experiment conducted earlier, fig plants formed by cuttings treated with gamma ray were selected based on their agronomic characteristics of interest. We determined the genetic variability in these fig tree selections, using RAPD and AFLP molecular markers, comparing them to each other and to the Roxo-de-Valinhos, used as the standard. For the reactions of DNA amplification, 140 RAPD primers and 12 primer combinations for AFLP analysis were used. The selections did not differ genetically between themselves and between them and the Roxo-de-Valinhos cultivar. Techniques that can detect polymorphism between treatments, such as DNA sequencing, must be tested. The phenotypic variation of plants may be due to epigenetic variation, necessitating the use of techniques with methylation-sensitive restriction enzymes.

Geographical variation and quality assessment of Stryphnodendron adstringens (Mart.) Coville within Brazil

Stryphnodendron adstringens (Mart.) Coville belongs to the Mimosoideae subfamily of the Leguminosae, and is also known as Barbatimão. It is extensively used in the traditional Brazilian medicine for its biological properties, which are attributed to the tannin fraction found in the bark. To maintain the genetic diversity of S. adstringens we have accessed twelve populations to establish an exsitu germplasm bank for future collections of seeds. Records on geographical boundary of occurrence, fruits, seeds and levels of extractions were gathered as species descriptors. Aiming the tannin fractions, we have noticed extraction of the bark on 46% of the evaluated trees as a practice that partially removes the bark disrupting wood vessels and causing premature death of the trees. In addition, the quality of seeds was low due to the high incidence of insect attack and immature abortion which compromises the species reproductive pattern. The geographical variation on the tannin content, seed production, predation and abortion was studied in twelve distinct populations located in three different states São Paulo, Goiás and Minas Gerais. Populations with the highest content of phenols and tannins were Campo Alegre and Luislândia. Our data demonstrated that there is a need for conservation of S. adstringens. Plants of the population located in Campo Alegre and Luislândia were considered elite for non-timber products of S. adstringens.