rastreamento histoquímico, fitoquímico e biológico de folhas de Plinia cauliflora (DC.) Kausel (Myrtaceae)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Anti-Candida targets and cytotoxicity of casuarinin isolated from Plinia cauliflora leaves in a bioactivity-guided study

In addition to the bio-guided investigation of the antifungal activity of Plinia cauliflora leaves against different Candida species, the major aim of the present study was the search for targets on the fungal cell. The most active antifungal fraction was purified by chromatography and characterized by NMR and mass spectrometry. The antifungal activity was evaluated against five Candida strains according to referenced guidelines. Cytotoxicity against fibroblast cells was determined. The likely targets of Candida albicans cells were assessed through interactions with ergosterol and cell wall composition, porosity and architecture. The chemical major component within the most active antifungal fraction of P. cauliflora leaves identified was the hydrolysable tannin casuarinin. The cytotoxic concentration was higher than the antifungal one. The first indication of plant target on cellular integrity was suggested by the antifungal activity ameliorated when using an osmotic support. The most important target for the tannin fraction studied was suggested by ultrastructural analysis of yeast cell walls revealing a denser mannan outer layer and wall porosity reduced. It is possible to imply that P. cauliflora targeted the C. albicans cell wall inducing some changes in the architecture, notably the outer glycoprotein layer, affecting the cell wall porosity without alteration of the polysaccharide or protein level. © 2013 by the authors.

Estudo da composição química, citotoxicidade e alvos da atividade antifúngica de Melaleuca alternifolia Cheel (Myrtaceae) e de Plinia cauliflora (Mart.) Kausel (Myrtaceae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Predicting soluble solid content in intact jaboticaba [Myrciaria jaboticaba (Vell.) O. Berg] fruit using near-infrared spectroscopy and chemometrics

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Avaliação da atividade antifúngica de extrato de Plinia cauliflora e Endopleura uchi

O uso indiscriminado dos medicamentos sintéticos e a própria evolução dos micro-organismos selecionou espécies extremamente resistentes aos agentes químicos utilizados. Para contornar tal situação e ampliar o arsenal de compostos ativos contra micro-organismos, o estudo de plantas tornou-se uma necessidade crescente. A utilização de fitoterápicos na prevenção e/ou cura de doenças são necessários estudos prévios relativos a aspectos botânicos, farmacognósticos, fitoquímicos, farmacológicos e toxicológicos. Neste trabalho, foram estudados extratos obtidos com as folhas de Plinia cauliflora (Mart.) Kausel e de cascas pulverizadas de Endopleura uchi (Huber) Cuatrec., quanto às propriedades antimicrobianas frente a diferentes linhagens de fungos: Aspergillus niger, Beauveria bassiana, Metarhizium anisopliae, Trichoderma reesei e Trichophyton rubrum. Foram avaliadas as atividades antifúngicas dos materiais vegetais determinando sua Concentração Inibitória Mínima (CIM). Com base nesses dados foi realizada a avaliação da atividade antifúngica observando o crescimento radial dos fungos em placas de Petri contendo meio de cultura incorporado com os materiais vegetais nas concentrações de CIM e 1%, sendo após realizada a mensuração dos diâmetros formados. Trichophyton rubrum foi o micro-organismo que apresentou maior sensibilidade frente ao extrato e as frações de P. cauliflora, com valores de CIM baixos, enquanto que a cepa industrial de Aspergillus niger mostrou-se totalmente resistente a todos os extratos testados. A determinação da atividade antifúngica em placas contendo meio sólido, mostrou uma confirmação dos resultados pois o extrato etanólico, a fração butanólica e a fração aquosa de P. cauliflora, tanto na concentração 1% quanto na obtida pelo CIM, inibiram o crescimento T. rubrum

Potential dietary sources of ellagic acid and other antioxidants among fruits consumed in Brazil: Jabuticaba (Myrciaria jaboticaba (Vell.) Berg)

BACKGROUND: The objectives of this study were (1) to evaluate the content of ellagic acid in fruits consumed by the Brazilian population, including native ones; (2) to further characterize rich sources in relation to ascorbic acid, phenolics contents and in vitro antioxidant capacity; and (3) to study the distribution and effect of ripening stage on ellagitannins content of jabuticaba (Myrciaria jaboticaba). The content of free ellagic acid and ellagic acid derivatives such as ellagitannins was analyzed using high-performance liquid chromatography (HPLC). RESULTS: Ellagic acid was detected in 10 out of a total of 35 fruits analyzed. The content of free ellagic acid in fruits varied from 0.0028 to 0.085 g kg(-1) (FW) and total ellagic acid varied from 0.215 to 3.11 g kg(-1) (FW). All the seven fruits belonging to the Myrtaceae family evaluated in this study presented high contents of ellagitannins in their composition, with jabuticaba, grumixama and cambuci (all native from Brazil) showing the highest total ellagic acid contents. Jabuticaba, the most consumed in Brazilamongthose and already adapted to commercial plantations, contained concentrated phenolics compounds, including ellagitannins, in the peel. Anthocyanins (cyanidin derivatives) increased significantly through ripening of jabuticaba and were not present in the pulp or seeds. Samples collected from three different locations during summer, winter and spring had total ellagic contents varying from 1.88 to 3.31 g kg(-1) (FW). The decrease in ellagic acid content with ripening was more accentuated for pulp (eight times) compared to seeds (2.3 times) and peel (2.0 times). CONCLUSION: These results showed the potential of jabuticaba as dietary source of ellagic acid and reinforced consumption of the whole fruit by the population. (C) 2011 Society of Chemical Industry

Características bioativas, funcionais e efeito protetor do resíduo desidratado de camu-camu (Myrciaria dubia H.B.K. (McVaugh)) sobre doenças degenerativas utilizando modelos in vivo C. elegans

Camu-camu (Myrciaria dubia H.B.K. (McVaugh)) is a native Amazon fruit, recognized worldwide as one of the main natural sources of ascorbic acid. Due to its great acidity, this fruit is generally consumed after processing into juice or as ingredient in food preparations. As a co-product of the camu-camu processing, a significant amount of agroindustrial residue is generated. Despite the studies showing the bioactive value and biological potential of the fruit, few studies have approached the possible processing techniques, transformation and preservation of camu-camu fruits and its agroindustrial pomace. Therefore, the present work has the objective of evaluating two different drying processes applied to camu-camu pomace (peel and seeds with residual pulp), freeze drying and hot air drying, in order to obtain a functional fruit product. This thesis was divided into three stages: the first one shows the studies related to the freeze drying and hot air drying, where we demonstrated the impact of the selected drying techniques on the bioactive components of camu-camu, taking the fresh pomace as the control group. Among the investigated conditions, the groups obtained at 50ºC and 4 m/s (SC50) and 80ºC and 6 m/s (SC80) were selected as for further studies, based on their ascorbic acid final content and Folin-Ciocalteau reducing capacity. In addition to SC50 and SC80, the fresh pomace (RF) and freeze dried (RL) samples were also evaluated in these further stages of the research. Overall, the results show higher bioactive concentration in the RF samples, followed by RL, SC50 and SC80. On the second step of the research, the antioxidant, antimicrobial and antienzymatic activities were evaluated and the same tendency was observed. It was also reported, for the first time in the literature, the presence of syringic acid in dried camu-camu pomace. In the third and final stage of the research, it was investigated the effect of dried camu-camu on aging and neuroprotective disorders, using the in vivo model C.elegans. It was observed that camu-camu extracts were able to modulate important signaling genes relevant to thermal and oxidative stresses (p < 0.05). The polar acid, polar basic and polar neutral fractions obtained from the low molecular extracts of SC50 were able to extend the lifespan of wild type N2 C. elegans in 20% and 13% (p < 0.001). Results also showed that the paralysis induced by the β1-42 amyloid was significantly (p < 0.0001) retarded in CL4176 worms. Similarly, the camu-camu extracts attenuated the dopaminergic induction associated to Parkinson’s disease. Finally, a global analysis of the data presented here reveal that the camu-camu pomace, a co-product obtained from the industrial processing of a native Brazilian fruit, is a relevant natural source of health relevant compounds. This thesis, shows for the first time, the multifunctionality of camu-camu pomace, a natural resource still underexploited for scientific, commercial and technological purposes.

Apomixia determinando a estrutura genética de uma população de Plinia cauliflora no sudoeste do Paraná

CAPES

Crescimento e desenvolvimento de Plinia cauliflora de acordo com a intensidade luminosa em clima subtropical

The jabuticaba fruit tree from classified in the Myrtaceae family and Plinia genre. There are about nine species of this fruit tree, that include as most important, Plinia trunciflora (jabuticaba de cabinho), naturally occurring in southwestern Paraná State, Brazil, P. cauliflora (jabuticaba Paulista or Jabuticaba Açu) and P. jaboticaba (Vell) (jabuticaba sabará), with all the over species producing fruit for the industry or fresh consumption. Nevertheless, there aren‟t commercial orchards with this culture, with highest yield part from extractive. This fact can be combined with lack of technical knowledge for the plants produce in the field. As these species are found in the forest, the first point is whether they can adapt to other light intensity conditions. The aim of this work was to identify the adaptive behavior of jabuticaba fruit seedling and tree when they were put in different light intensities and what this can be considered ideal for the growth, as well as, its influence in the leaves secondary compounds production. Two experiments were conducted, with the first involved with the study of the seedlings and the second with plants in the field. The work was carried out at Universidade Tecnológica Federal do Paraná – Câmpus Dois Vizinhos, Paraná State - Brazil. The experimental design was a completely randomized and a block design with four treatments and four replications of 10 seedlings or two plants per plot, according to nursery or orchard conditions, respectively. The treatments were base according to the light intensity. The treatments used were, 1 - full sun, similar the orchard condition, with 0% shading; 2 - side cover with shade cloth and top with transparent plastic, representing a gap forest condition; 3 - side and top cover with shade cloth, representing stage where the forest canopy is closing, focusing only indirect sunlight; 4 - side and top cover with shade cloth, simulating a closed canopy condition, with PPD (photon flux density) of 10% (90% shading); 5 - side and top cover with shade cloth, simulating a more open canopy condition with PPD 65% (35% shading). The growth and development seedling and plant characteristics were evaluated once by month, as also, during time part in the plants the secondary metabolites leaves, soil activity microbiological and the fresh and dry matter root and shoot and, root length from seedlings. For the growth and development of jabuticaba Açú Paulista seedling recommend to use of side cover with shade cloth and top with transparent plastic, representing a gap forest condition. In orchard, for the growth and development of plants jabuticaba Híbrida tree it was recommended the use of side and top cover with shade cloth of some type. For production of secondary metabolites of leaves, the plant must to be full sunlight condition orchard.

Desempenho vegetativo de diferentes acessos de camucamuzeiro (Myrciaria dubia (H.B.K.) Mc Vaugh) cultivados em terra firme

A Amazônia se constitui enorme fonte de biodiversidade não utilizada pela humanidade. Um exemplo desta biodiversidade é o camucamuzeiro (Myrciaria dubia (H.B.K.) Mc Vaugh), que atualmente desperta grande interesse devido ao potencial de produção de frutos com alto teor de ácido ascórbico, quantidade esta que varia de 2 400 a 3 000 mg/100g de mesocarpo a ate 5 000 mg /100g de casca. O trabalho teve como objetivo avaliar o desempenho vegetativo de 11 acessos coletados em populações naturais e cultivados em terra firme. Os acessos foram coletados no rio Solimões e plantados em Campo Experimental da Embrapa Amazônia Oriental, em Belém, Para, em Latossolo Amarelo textura média, delineamento em blocos ao acaso com duas repetições. As unidades experimentais foram constituídas de cinco planta úteis e competitivas, plantadas em espaçamento de 4m x 4m, com bordadura simples nas extremidades. Foram avaliados dados de altura da planta, diâmetro do caule principal a 60 cm do solo, número de caules, comprimento e largura da folha e área foliar. Detectaram-se diferenças significativas (p<0,05) para diâmetro do caule e comprimento da folha, e altamente significativas (p<0,01) para número de caules, largura da folha e área foliar, indicando que estes são caracteres capazes de discriminar esses acessos. Os resultados demonstraram que é possível cultivar camucamuzeiro em terra firme e que o acesso Solimões 1012 apresentou melhor desempenho nas condições estudadas. Conclui-se que a população Solimões 1012 poderá ser utilizada quando da implantação de pomares de camucamuzeiro.

Germinação in vitro de camu-camuzeiro (Myrciaria dubia (H.B.K.) McVaugh).

O sucesso da aplicação das técnicas de cultura de tecidos é dependente do estabelecimento prévio da cultura que se quer estudar in vitro. Para a produção de plântulas assépticas, um dos principais problemas é o alto índice de contaminação de explantes obtidos diretamente de plantas de campo. Este trabalho objetivou avaliar a percentagem de germinação de sementes de camu-camuzeiro, em dois experimentos de germinação in vitro, visando a obtenção de plântulas assépticas. No primeiro experimento foram utilizadas cinco concentrações diferentes de ácido giberélico (AG3) combinadas a cinco tipos de explantes obtidos a partir de sementes maduras de camu-camuzeiro, em um delineamento experimental inteiramente casualizado em esquema fatorial 5x5. No segundo experimento, o delineamento experimental foi inteiramente casualizado em esquema fatorial 2x3x4, utilizando ausência e presença de carvão ativado a 3g.L-1, sementes em três estádios de maturação e quatro concentrações de AG3 (0; 1; 2 e 4mg.L-1). Os explantes foram inoculados em meio MS com pH 5,8, contendo 20g.L-' de sacarose e 2g.L-1 de phytagel. A incubação foi feita em uma sala com fotoperíodo de 16h/luz/dia e temperatura de 25° ± 2°C, e a característica avaliada foi a percentagem de germinação. Nos dois experimentos, o número de explantes germinados foi baixo e independente do estádio de maturação, da presença de AG3 e de carvão ativado no meio de cultura.

Bioactive compounds and antioxidant capacities of 18 non-traditional tropical fruits from Brazil

The bioactive compounds and antioxidant capacities of polyphenolic extracts of 18 fresh and dry native non-traditional fruits from Brazil were determined using ABTS, DDPH, FRAP and beta-carotene bleaching methods. The study provides an adaptation of these methods, along with an evaluation of the compounds related to antioxidant potential. The results show promising perspectives for the exploitation of non-traditional tropical fruit species with considerable levels of nutrients and antioxidant capacity. Although evaluation methods and results reported have not yet been sufficiently standardised, making comparisons difficult, our data add valuable information to current knowledge of the nutritional properties of tropical fruits, such as the considerable antioxidant capacity found for acerola - Malpighia emarginata and camu-camu - Myrciaria dubia (ABTS, DPPH and FRAP) and for puca-preto - Mouriri pusa (all methods). (C) 2010 Elsevier Ltd. All rights reserved.

Bioactive compounds and antioxidant capacity of exotic fruits and commercial frozen pulps from Brazil

The objective of this work was to characterize exotic fruits (cambuci, araca-boi, camu-camu, jaracatia, araca) and commercial frozen pulps (araca, cambuci, umbu, coquinho, pana, native passion fruit, cagaita) from Brazil in relation to their bioactive compounds contents and antioxidant capacity. Camu-camu (Myrciaria dubia) presented the highest vitamin C and total phenolics contents (397 and 1797 mg/100 g f.w., respectively) and the highest DPPH(center dot) scavenging capacity. Coquinho (Butia capitata) also showed a significant vitamin C content (43 mg/100 g f.w.). Among the commercial frozen pulps, cagaita presented the higher DPPH scavenging activity and inhibition of beta-carotene bleaching. A good correlation between total phenols and DPPH scavenging activity was found for fruits (r = 0.997) and commercial frozen pulps (r = 0.738). However, no correlation was found for total phenols and inhibition of beta-carotene bleaching. Quercetin and kaempferol derivatives were the main flavonoids present in all samples and cyanidin derivatives were detected only in camu-camu. Camu-camu and araca (Psidium guineensis) showed the highest total ellagic acid contents (48 and 63.5 mg/100 g f.w.). All commercial frozen pulps presented lower contents of bioactive compounds and antioxidant capacity than their respective fruits. According to our results, camu-camu and araca might be sources of bioactive compounds.

Pré-melhoramento do camucamuzeiro: estudo de parâmetros genéticos e dissimilaridade

O camucamuzeiro pertence à família Myrtaceae e é espécie em processo de domesticação, encontrada na forma extrativa, a partir de plantas crescendo naturalmente nas margens dos rios e lagos, ou cultivadas em pequenas áreas de terra firme. As estimativas e a compreensão dos parâmetros genéticos desta espécie são importantes para o conhecimento da estrutura genética das populações e para a inferência da diversidade genética presente, além de proporcionar subsídios para predizer os ganhos genéticos e o possível sucesso no programa de melhoramento dessa cultura. Neste sentido, o objetivo deste trabalho foi estimar parâmetros genéticos e a dissimilaridade genética, em acessos existentes no Banco Ativo de Germoplasma de camucamuzeiro, da Embrapa Amazônia Oriental. Para o estudo, foram analisados 46 progênies, colhidos 40 frutos por planta matriz em completo estádio de maturação (frutos com epicarpo totalmente roxo), sendo avaliados sete caracteres morfoagronômicos: peso de fruto (g, PFR), comprimento de fruto (cm, CFR), diâmetro de fruto (cm, DFR), peso da casca (g, PCS), espessura da casca (cm, ECS), número de sementes (n, NSE), peso de sementes (g, PSE). Por meio do Programa Genes, estimaram-se os componentes de variância, herdabilidade e a variabilidade. A importância relativa de caracteres e dissimilaridades entre as progênies, bem como as correlações genéticas entres os caracteres avaliados também foram estudadas. Verificou-se que há dissimilaridade entre os acessos do BAG de camucamuzeiro e que, por causa das correlações significativas entre as variáveis, podem-se adotar métodos de seleção indireta como ferramenta auxiliar no processo de domesticação e melhoramento desta espécie.