1H HR-MAS NMR and chemometric methods for discrimination and classification of Baccharis (Asteraceae): A proposal for quality control of Baccharis trimera.

Baccharis trimera is a species recognized by health agencies and recommended by the Brazilian Pharmacopoeia by having medicinal properties. In this work, HR-MAS NMR spectroscopy combined with chemometric tools, such as Principal Components Analysis (PCA), Partial Least Squares Discriminant Analysis (PLS-DA) and Soft Independent Modeling by Class Analogy (SIMCA), were used to evaluate the quality control and authenticity of commercial samples of Baccharis, as well as to discriminate B. trimera samples from other species of the Caulopterae section (B. articulata, B. trimera, B. junciformis, B. milleflora, and B. myriocephala). The high morphological similarity of these species makes it difficult for their identifications and discriminations, even by taxonomists. Different PCA pre-processing (autoscaling, Pareto scaling, and mean centering) allowed to discriminate B. trimera and B. myriocephala from the other species, mainly due to the presence of carquejyl acetate, indicated their chemical similarity. The 1H HR-MAS NMR spectral profile offers the possibility of tracking not only the chemical markers, such as the presence of carquejyl acetate, which can also be helpful in the B. trimera authentication/identification. The application of classification methods in standard samples revealed that PLS-DA models showed better performance on the calibration and validation sets than SIMCA model. However, PLS-DA and SIMCA applied to commercial samples showed that none of the commercial samples were classified as B. trimera, which suggested the lack of strict quality control regarding these products. The methodology developed in the present work might contribute to chemotaxonomy of the genus Baccharis.

Essential Oils of Five Baccharis Species: Investigations on the Chemical Composition and Biological Activities.

This paper provides a comparative account of the essential oil chemical composition and biological activities of five Brazilian species of Baccharis (Asteraceae), namely B. microdonta, B. pauciflosculosa, B. punctulata, B. reticularioides, and B. sphenophylla. The chemical compositions of three species (B. pauciflosculosa, B. reticularioides, and B. sphenophylla) are reported for the first time. Analyses by GC/MS showed notable differences in the essential oil compositions of the five species. α-Pinene was observed in the highest concentration (24.50%) in B. reticularioides. Other major compounds included α-bisabolol (23.63%) in B. punctulata, spathulenol (24.74%) and kongol (22.22%) in B. microdonta, ß-pinene (18.33%) and limonene (18.77%) in B. pauciflosculosa, and ß-pinene (15.24%), limonene (14.33%), and spathulenol (13.15%) in B. sphenophylla. In vitro analyses for antimalarial, antitrypanosomal, and insecticidal activities were conducted for all of the species. B. microdonta and B. reticularioides showed good antitrypanosomal activities; B. sphenophylla showed insecticidal activities in fumigation bioassay against bed bugs; and B. pauciflosculosa, B. reticularioides, and B. sphenophylla exhibited moderate antimalarial activities. B. microdonta and B. punctulata showed cytotoxicity. The leaves and stems of all five species showed glandular trichomes and ducts as secretory structures. DNA barcoding successfully determined the main DNA sequences of the investigated species and enabled authenticating them.

Screening antimycobacterial activity of Baccharis dracunculifolia, Centella asiatica, Lantana camara and Pterodon emarginatus / Atividade anti-micobacteriana de Baccharis dracunculifolia, Centella asiatica, Lantana camara e Pterodon emarginatus

ABSTRACT The permanent investigation of new antimycobacterial drugs is necessary for the eradication programs of tuberculosis and other mycobacterium-related diseases. The aim of the present study is to search for new sources of antimycobacterial drugs using plant materials. In this study, 11 plant materials (extracts, essential oils and some fractions) obtained from 4 species of medicinal plants traditionally used as general therapeutics for different illnesses and specifically as treatment of tuberculosis, were evaluated using the microplate resazurin assay against 2 species of the Mycobacterium tuberculosis Complex and 3 nontuberculous mycobacteria. The results showed the hexane extract and the essential oil from fruits of Pterodonemarginatus (Vogel) as potential sources of antimycobacterial drugs against 4 species of tested mycobacteria. The hexane fraction of methanol extract from leaves of Centella asiatica also presented significant mycobacterial growth inhibition, but against M. chelonae only. In conclusion, it was possible to contribute to the antimycobacterial investigations by presenting three new samples of plants with significant antimicrobial activity against four Mycobacteriumspp and suggest future studies about the antimycobacterial properties of fruits from P. emarginatus.

RESUMO A investigação permanente de novas drogas antimicobacterianas é necessária no programa de erradicação da tuberculose e de outras doenças relacionadas com micobactérias. O objetivo deste estudo foi buscar novas fontes de drogas antimicobacterianas usando material vegetal. Neste estudo, 11 materiais de base vegetal (extratos, óleos essenciais e algumas frações) foram avaliados contra 5 espécies de micobactérias. Estes materiais foram obtidos a partir de 4 espécies de plantas medicinais tradicionalmente utilizadas como terapêutica geral para diferentes doenças e, especificamente, no tratamento de tuberculose (Baccharis dracunculifolia, Centella asiatica, Lantana camara, Pterodon emarginatus). Os ensaios foram realizados em microplacas com resazurina contra duas espécies do Complexo Mycobacteriumtuberculosis e 3 espécies de micobactérias não tuberculosas. Os resultados mostraram o extrato hexânico e o óleo essencial de frutos de P.emarginatus como potenciais fontes para drogas antimicobacterianas contra quatro espécies de micobactérias testadas. A fração hexânica do extrato metanólico das folhas de C. asiatica também apresentou significativa inibição do crescimento de micobactérias apenas contra M.chelonae. Em conclusão, foi possível contribuir para as investigações de antimicobacterianos por apresentar três novas amostras de plantas com atividade antimicrobiana significativa contra quatro Mycobacterium spp e sugerir a realização de estudos futuros sobre as propriedades antimicobacterianas de frutos de P. emarginatus.

Antioxidant properties of phenolic compounds from Baccharis articulata and B. usterii.

As a continuation of our research on the Baccharis genus, the evaluation of the antioxidant effect by TRAP and TBARS assays of three isolated compounds from n-butanol fractions of B. articulata and B. usterii is reported. The structures of these compounds were established as 4-O-beta-D-glucopyranosyl-3,5-dimethoxybenzyl-methanol (1), 5-O-[E]-dicaffeoylquinic acid (2), and 7-hydroxy-5,4'-dimethoxyflavone (3). In the TRAP assay it was possible to observe an antioxidant effect of both n-butanol fractions at 1.25 microg/mL. Among the isolated compounds, compound 2 displayed a remarkable contribution to the total antioxidant capacity of the n-butanol fraction of B. usterii. Moreover, the n-butanol fractions of both species, compounds 1 and 2 at 40.0 microg/mL were efficient in protection of lipid peroxidation in the TBARS experiment. They are promising lead compounds for use in medicinal chemistry studies.

Influence of plant genetic diversity on interactions between higher trophic levels.

While the ecological consequences of plant diversity have received much attention, the mechanisms by which intraspecific diversity affects associated communities remains understudied. We report on a field experiment documenting the effects of patch diversity in the plant Baccharis salicifolia (genotypic monocultures versus polycultures of four genotypes), ants (presence versus absence) and their interaction on ant-tended aphids, ants and parasitic wasps, and the mechanistic pathways by which diversity influences their multi-trophic interactions. Five months after planting, polycultures (versus monocultures) had increased abundances of aphids (threefold), ants (3.2-fold) and parasitoids (1.7-fold) owing to non-additive effects of genetic diversity. The effect on aphids was direct, as plant genetic diversity did not mediate ant-aphid, parasitoid-aphid or ant-parasitoid interactions. This increase in aphid abundance occurred even though plant growth (and thus aphid resources) was not higher in polycultures. The increase in ants and parasitoids was an indirect effect, due entirely to higher aphid abundance. Ants reduced parasitoid abundance by 60 per cent, but did not affect aphid abundance or plant growth, and these top-down effects were equivalent between monocultures and polycultures. In summary, intraspecific plant diversity did not increase primary productivity, but nevertheless had strong effects across multiple trophic levels, and effects on both herbivore mutualists and enemies could be predicted entirely as an extension of plant-herbivore interactions.

Baccharis rufescens Spreng. var. tenuifolia (DC. ) Baker: contribuição ao estudo farmacognóstico / Baccharis rufescens Spreng. var. tenuifolia (DC. ) Baker: contribution to the pharmacognostic study

Baccharis rufescens Spreng. var. tenuifolia (DC.) Baker pertence à família Asteraceae e é usada na medicina tradicional como estomáquico e hepatoprotetor. Um estudo fitoquímico mostrou a presença de flavonoides e triterpenos, sendo que os extratos clorofórmico e metanólico de folhas apresentaram-se ativos no bioensaio de toxicidade sobre Artemia salina Leach. Além disso, extratos clorofórmicos evidenciaram a presença de peróxidos, sugerindo sua aplicação no tratamento da malária (Schenkel at al., 2002; Montanher et al., 2002; Moreira et al., 2003). Considerando a importância farmacológica de B. rufescens var. tenuifolia, o presente trabalho objetivou o estudo morfoanatômico e histoquímico do caule e da folha dessa espécie, a fim de fornecer subsídios farmacognósticos para o controle de qualidade. O material botânico foi submetido às técnicas usuais empregadas na microscopia de luz e microscopia eletrônica de varredura. Folha anfiestomática, presença de estômatos anomocíticos, tricomas glandulares capitados bisseriados, tricomas tectores flageliformes simples unisseriados, dutos secretores associados ao floema, calota de fibras perivasculares e cristais de oxalato de cálcio do tipo estiloide e prismático na região medular do caule foram as principais características observadas que auxiliam na identificação do táxon.

Baccharis rufescens Spreng. var. tenuifolia (DC.) Baker belongs to the Asteraceae family. It is used for liver and stomach problems in traditional medicine. Previous phytochemical data reported flavonoid and triterpene contents. Chloroform and methanol extracts of the leaves showed activity in the bioassay of brine shrimp. Chloroform extracts showed the presence of peroxides that can be used to treat malaria (Schenkel et al., 2002; Montanher et al., 2002; Moreira et al., 2003). Considering the pharmacological importance of B. rufescens var. tenuifolia, the purpose of this paper was to perform the anatomical analysis of aerial vegetative parts of Baccharis rufescens var. tenuifolia in order to provide pharmacognostic data for quality control. The plant material was studied by the usual methods of light and scanning electron microscopy. Amphistomatic leaves, anomocytic stomata, biseriate capitate glandular trichomes, uniseriate simple flagelliform non-glandular trichomes, secretory ducts associated to the phloem, perivascular fiber cap, calcium oxalate as prismatic and styloid crystals in the pith of the stem were reported as the mainly anatomical data for B. rufescens var. tenuifolia.Ouvir Ler foneticamente.

Interaction between Apis mellifera L. and Baccharis dracunculifolia DC, that favours green propolis production in Minas Gerais.

In Minas Gerais, green propolis is produced from the collection of resinous substance found in shoot apices of Baccharis dracunculifolia. The aim of this study was to evaluate the biological parameters associated with the interaction Apis mellifera x Baccharis dracunculifolia, to elucidate the supply of resin for green propolis production in Minas Gerais. We selected male and female individuals of two populations of Baccharis dracunculifolia located on São Judas Tadeu Farm - FSJT, in the municipality of Betim, MG and the Experimental Garden of the Ezequiel Dias Foundation - HORTO, located in an urban area in Belo Horizonte, MG. We made weekly observations, from June 2007 to June 2008, and evaluated in both populations: richness and abundance of insect visitors; resin collecting visits of Apis mellifera; presence of Baccharopelma dracunculifoliae galls; growth of individuals and phenological phases. Statistical analyses were made using R software. The rainy season showed the highest number of visitors. A. mellifera collected resin in shoot apices of Baccharis dracunculifolia from August to April, only in the FSJT population, where galls of B. dracunculifoliae were also present. Ovoposition of gall inductor on host plants occurs during the rainy season, when there is a peak of visitants and resin collecting visits of honeybees. This fact stimulates plant defense strategies against parasitoids and predators, which includes the production of several secondary metabolites, and ultimately reduces competition for food by inhibiting the attack of other phytophagous insects, not adapted to the chemical environment of plant tissues. Green propolis production in Minas Gerais is related to the abundant supply of resin by Baccharis dracunculifolia, when they are parasitised by B. dracunculifoliae galls. They induce plant production of defense exudates, which attract Apis mellifera bees to collect resin and consequently favour the production of green propolis.

Interaction between Apis mellifera L. and Baccharis dracunculifolia DC, that favours green propolis production in Minas Gerais / Interação entre Apis mellifera L. e Baccharis dracunculifolia DC que favorecem a produção de própolis verde no estado de Minas Gerais

In Minas Gerais, green propolis is produced from the collection of resinous substance found in shoot apices of Baccharis dracunculifolia. The aim of this study was to evaluate the biological parameters associated with the interaction Apis mellifera x Baccharis dracunculifolia, to elucidate the supply of resin for green propolis production in Minas Gerais. We selected male and female individuals of two populations of Baccharis dracunculifolia located on São Judas Tadeu Farm - FSJT, in the municipality of Betim, MG and the Experimental Garden of the Ezequiel Dias Foundation - HORTO, located in an urban area in Belo Horizonte, MG. We made weekly observations, from June 2007 to June 2008, and evaluated in both populations: richness and abundance of insect visitors; resin collecting visits of Apis mellifera; presence of Baccharopelma dracunculifoliae galls; growth of individuals and phenological phases. Statistical analyses were made using R software. The rainy season showed the highest number of visitors. A. mellifera collected resin in shoot apices of Baccharis dracunculifolia from August to April, only in the FSJT population, where galls of B. dracunculifoliae were also present. Ovoposition of gall inductor on host plants occurs during the rainy season, when there is a peak of visitants and resin collecting visits of honeybees. This fact stimulates plant defense strategies against parasitoids and predators, which includes the production of several secondary metabolites, and ultimately reduces competition for food by inhibiting the attack of other phytophagous insects, not adapted to the chemical environment of plant tissues. Green propolis production in Minas Gerais is related to the abundant supply of resin by Baccharis dracunculifolia, when they are parasitised by B. dracunculifoliae galls. They induce plant production of defense exudates, which attract Apis mellifera bees to collect resin and consequently favour the production of green propolis.

A própolis verde é produzida pelas abelhas Apis mellifera no estado de Minas Gerais, Brasil, a partir da coleta de substância resinosa dos ápices vegetativos de Baccharis dracunculifolia. O objetivo deste trabalho foi avaliar os parâmetros biológicos associados à interação Apis mellifera x Baccharis dracunculifolia, visando elucidar o fornecimento de resina da espécie para a produção de própolis verde em Minas Gerais. Foram selecionados indivíduos masculinos e femininos em duas populações de Baccharis dracunculifolia localizadas na Fazenda São Judas Tadeu – FSJT, município de Betim, MG, e no Horto Experimental da Fundação Ezequiel Dias – HORTO, em área urbana no município de Belo Horizonte, MG. Foram observados semanalmente, entre junho de 2007 e junho de 2008, a riqueza e abundância de insetos visitantes; visitação de Apis mellifera para coleta de resina; presença de galhas de Baccharopelma dracunculifoliae; crescimento dos indivíduos de Baccharis dracunculifolia e fenofases. Para análise estatística utilizou-se o software R. Em ambas as populações, a estação chuvosa foi a que apresentou maior número de visitantes. As abelhas Apis mellifera, coletaram resina nos ápices vegetativos de Baccharis dracunculifolia entre agosto e abril, e somente na população FSJT, onde ocorreram galhas de B. dracunculifoliae. Neste período a planta é ovopositada pelo indutor da galha que estimula a produção de diversos metabólitos secundários, como proteção ao ataque de parasitoides e predadores, e reduz a competição por alimento ao inibir o ataque de outros insetos fitófagos. A produção de própolis verde em Minas Gerais está relacionada à oferta abundante de resina pela planta Baccharis dracunculifolia, parasitadas por essas galhas. A atração das abelhas Apis mellifera para coleta de resina e produção de própolis verde se dá pela produção de exsudatos de defesa pela planta induzida pela presença da galha.

Analysis of propolis from Baccharis dracunculifolia DC. (Compositae) and its effects on mouse fibroblasts.

This paper confirms Baccharis dracunculifolia DC. (Compositae) as the main botanical source of the propolis from southeastern Brazil (state of São Paulo) investigated to ascertain specific biological activity in relation to mouse NIH-3T3 fibroblasts, skin cells directly involved in the cicatrization processes. Flavonoid and total phenolic compounds were determined by spectrophotometry, and chemical composition by HPLC; the chromatographic profile, characterized largely by flavonoids and aromatic acids, was found to be qualitatively similar to that of Baccharis dracunculifolia DC. The adsorption of phenolic compounds in the propolis to skin powder was also investigated, and 68% of these compounds adsorbed to the skin powder. At concentrations from 0.12 to 7.81 microg/ml, the propolis revealed no statistical significant differences from its control solutions; however, at concentrations of 31.25 microg/ml or more, the propolis was toxic to NIH-3T3 cells. Thus, the propolis from Baccharis dracunculifolia DC. (Compositae) presents an in vitro concentration-dependent toxicity on mouse NIH-3T3 fibroblasts.

Numerical taxonomy characterization of Baccharis genus species by ultraviolet-visible spectrophotometry.

Numerical taxonomy characterization of Baccharis genus species was performed using ultraviolet-visible spectrophotometry. The aim was to present a more convenient, more practical, more economic and faster method based on chemometric methods and UV-vis absorbance to give the most information about species identity and discrimination, especially when their classification has been doubtful. Three Baccharis species: B. genistelloides Persoon var. trimera (Less.) DC, B. milleflora (Less.) DC, and B. articulata (Lam.) Persoon were included in the study. With the help of principal component analysis (PCA) and cluster analysis (CA), we could characterize the three species. Application of soft independent modeling of class analogy (SIMCA) and K-nearest neighbor (KNN) methods on a training set of 65 extracts resulted in models that correctly classified all samples of an independent validation set, eight samples of B. genistelloides Persoon var. trimera (Less.) DC and one sample donated by Prof. Alarich Schultz Herbarium, Porto Alegre-RS, Brazil.

Differentiation of species of the Baccharis genus by HPLC and chemometric methods.

Phytochemical investigation of the aerial parts of three Baccharis species (Asteraceae family) was performed using HPLC and chemometric methods, with the objective of distinguishing between three morphologically very similar species: Baccharis genistelloides Persoon var. trimera (Less.) DC, B. milleflora (Less.) DC and B. articulata (Lam.) Persoon. With the help of Principal Component Analysis (PCA) and variance weights, it was possible to characterize the chromatographic profiles of the alcoholic extracts of the three species. Application of Soft Independent Modeling of Class Analogy (SIMCA) and K-Nearest Neighbor (KNN) methods on a training set of 74 extracts resulted in models that correctly classified all eight samples in an independent test set.