Abstract INTRODUCTION: Despite their widespread usage, synthetic insecticides and larvicides are harmful for controlling disease-causing mosquitoes owing to the development of resistance. The leaves of Eugenia astringens, Myrrhinium atropurpureum, and Neomitranthes obscura were collected from Marambaia and Grumari restingas. The safety and larvicidal efficacy of their extracts were tested against Aedes (Stegomyia) aegypti L. and Simulium (Chirostilbia) pertinax Kollar. METHODS: The dry leaves were subjected to static maceration extraction using 90% methanol. A. aegypti and S. pertinax larvae were exposed to 7.5, 12.5, and 25.0 µL/mL of the extracts (n= 30). The larvicidal activity after 24 h and 48 h, and the mortality, were determined. The median lethal concentration (CL50) was estimated by a Finney's probit model. RESULTS: M. atropurpureum and E. astringens extracts exhibited the strongest larvicidal effects against A. aegypti. M. atropurpureum extracts (25 µL/mL) caused mortalities of over 50% and 100% after 24 h and 48 h, respectively (CL50 = 11.10 and 9.68 ppm, respectively). E. astringens extracts (25 µL/mL) caused mortalities of 50% and 63.33% after 24 h and 48 h, respectively. High concentrations of N. obscura extracts induced a maximum mortality of 46.66% in A. aegypti larvae after 48 h (CL50= 25 ppm). The larvae of S. pertinax showed 100% mortality following exposure to all the plant extracts at all the tested concentrations after 24 h. CONCLUSIONS: The extracts of M. atropurpuerum exhibited the strongest larvicidal activity against A. aegypti. The larvae of S. pertinax were sensitive to all the extracts at all the tested concentrations.

Abstract Phenolic compounds are one of the main groups of secondary metabolites in plants and are known for their antioxidant activity. Rosmarinus officinalis L. (rosemary) contains different phenolic compounds including carnosol, carnosic acid and rosmarinic acid. In Cuba, rosemary cultivation is limited because it is difficult to propagate and has a low yield. As a result, it was removed from the Herbal Medicine National Formulary. However, the National Public Health System has a strong interest in rosemary because of its value as a natural antioxidant medicine. Irrigation with water treated with a static magnetic field (SMF) is a possible strategy to increase rosemary yield. This technology has been applied to accelerate plant growth and increase crop quality. The aim of this study was to evaluate the content of phenolic compounds and antioxidant activity in aqueous leaf extracts from plants irrigated with SMF-treated water in comparison to control plants. Significant differences in phenolic content and antioxidant activity were observed between aqueous extracts of control plants and plants irrigated with SMF-treated water. Therefore, irrigation with SMF-treated water is a promising technology to improve the cultivation of rosemary as a raw material to obtain pharmaceutical products with high antioxidant activities.

A alta demanda por material vegetal de Calendula officinalis para produção de fitoterápicos e cosméticos, configura a técnica de cultura de tecidos vegetais como uma das alternativas para o aprimoramento das culturas em curto período de tempo. Um protocolo de cultura de tecidos foi desenvolvido a partir de segmentos de plântulas de C. officinalis, no intuito de melhorar a proliferação de brotos e o enraizamento. Foi utilizado o meio Murashige e Skoog (MS½N) reduzido à metade das concentrações de NH4NO3 e KNO3, para verificar o efeito de diferentes tipos de explantes (basal, intermediário e apical), meio contendo areia de praia como suporte em substituição do agar, e o efeito de citocininas e auxinas (TDZ, tidiazuron; BAP, 6-benzilaminopurina; AIA, ácido indol-3-acético; AIB, ácido indol-3-butírico; ANA, ácido naftaleno acético) no desenvolvimento vegetal in vitro. Os resultados mostraram pronunciado enraizamento a partir dos explantes apicais, e também maior alongamento dos brotos e número de folhas. O meio sólido foi mais apropriado para as culturas de C. officinalis. A proliferação de brotos mostrou-se dependente de citocininas, com melhores resultados sob efeito de TDZ ou BAP em relação aos outros tratamentos. As plantas regeneradas a partir do meio contendo TDZ apresentaram aspecto vitrificado e deformações na morfogênese. A maior taxa de enraizamento (80%) foi obtida utilizando AIA 0.1 mg L-1. Através da propagação in vitro foram obtidas plantas de C. officinalis saudáveis e com raízes, capazes de serem aclimatizadas, e permitindo contínuo suplemento de matéria-prima.

The high demand for plant material from Calendula officinalis in the production of herbal medicines and cosmetics, turns the technique of plant-tissue culture into one of the alternatives for the improvement of crops over a short period of time. A protocol for tissue culture was developed from segments of seedlings of C. officinalis, in order to improve the proliferation of shoots and roots. We used a Murashige and Skoog (MS½N) medium, reduced to half the concentrations of NH4NO3 and KNO3 to verify the effect of different types of explants (basal, intermediate, and apical), a medium containing beach sand as support instead of agar, and the effect of auxins and cytokinins (TDZ tidiazuron; BAP, 6-benzylaminopurine, IAA, indol-3-acetic acid, IBA, indol-3-butyric acid, NAA, naphthalene-acetic acid) on plant development in vitro. The results showed pronounced rooting from the apical explants, as well as a greater elongation of shoots and number of leaves. The solid medium was more suitable for the C. officinalis cultures. Shoot proliferation was dependent on cytokinins with better results from the influence of TDZ or BAP compared to the other treatments. Plants regenerated from the medium containing TDZ displayed a glazed appearance and morphogenetic deformations. The highest rate for rooting (80%) was obtained using IAA 0.1mgL-1. Through in-vitro propagation, healthy C. officinalis plants were obtained with roots which can acclimatise, allowing the continuous supplement of raw material.

O estabelecimento de cultura in vitro de Polygala paniculata L. permitiu a propagação rápida de matéria-prima vegetal sob condições assépticas e padronizadas. As plantas in vitro foram submetidas à extração e destilação simultâneas (EDS) e a composição química do óleo essencial foi analisada por CG/DIC e CG/EM. Foram avaliados os efeitos de BAP (6-benzilaminopurina) sobre o desenvolvimento in vitro e produção de voláteis em P. paniculata. A principal característica do óleo essencial consistiu na presença marcante de salicilato de metila. Plantas cultivadas em meio controle MS (Murashige & Skoog) produziram mais do que 80% de salicilato, enquanto o uso de BAP, reduziu a produção de salicilato de metila a 71% e 21% para as concentrações de 2 e 4 mg L-1, respectivamente. A cultura de tecidos de P. paniculata pode ser indicada como fonte de salicilato de metila.

The establishment of Polygala paniculata L. tissue culture allowed a rapid propagation of raw plant material under aseptic and standardized conditions. Plantlets were subjected to simultaneous distillation-extraction (SDE) and volatiles composition analyzed by GC/FID and GC/MS. The effect of BAP (6-benzylaminopurine) on in vitro development and volatiles production was evaluated. The main feature of volatile compounds was the significant amount of methyl salicylate. Plantlets cultured on control MS (Murashige & Skoog) medium showed more than 80% of salicylate, while the addition of BAP at 2 and 4 mg L-1 concentrations reduced methyl salicylate production to 71% and 21%, respectively. Tissue culture of P. paniculata proved to be a potential source of methyl salicylate.

Volatiles produced by plantlets of Alpinia zerumbet were obtained by means of simultaneous distillation-extraction (SDE). The effects of indole-3-acetic acid, kinetin, thidiazuron and 6-benzylaminopurine on leaf and root volatile composition obtained by tissue cultures were investigated. A higher content of b-pinene and a lower content of sabinene were observed in leaf volatile of plantlets cultured in control, IAA and IAA+ TDZ media, as compared with those of donor plants. In vitro conditions were favorable to increase caryophyllene content. Volatile compounds from the root were characterized mainly by camphene, fenchyl-acetate and bornyl acetate; which constitute about 60% of total volatile.

Este estudo foi conduzido para investigar os efeitos dos espectros de luz, adição de UV-A, e diferentes concentrações reguladores de crescimento sobre a germinação in vitro de Senecio cineraria DC. As sementes foram desinfestadas e inoculadas em meio MS para avaliação das seguintes qualidades de luz: branca, branca acrescida de UV-A, azul, verde, vermelha ou ausência de luz. A maior germinabilidade foi obtida usando meio MS0 sob luz branca (30%) e MS + 0,3 mg L-1 de AG3 no escuro (30,5%). Sementes de S. cineraria foram indiferentes à luz para germinar. As luzes azul e verde inibiram a germinação. Diferentes concentrações de ácido giberélico (AG3) (0,1; 0,4; 0,6; 0,8; 1,0 e 2,0 mg L-1) e ácido indolacético (AIA) (0,1; 0,3 e 1,0 mg L-1) foram avaliadas sob luz branca e no escuro. Sob luz branca, nenhuma concentração de AG3 aumentou a taxa germinativa. As concentrações de AG3 aprimoraram as respostas de germinação para as sementes mantidas no escuro, exceto na concentração de 1,0 mg L-1. Essas concentrações também aumentaram o tempo médio e reduziram a velocidade de germinação. A taxa de germinação, sob condições de luz e no escuro, foi menor quando se utilizou AIA em comparação com AG3.

This study was carried out to investigate the effects of light spectra, additional UV-A, and different growth regulators on the in vitro germination of Senecio cineraria DC. Seeds were surface-sterilized and inoculated in MS medium to evaluate the following light spectra: white, white plus UV-A, blue, green, red or darkness. The maximum germinability was obtained using MS0 medium under white light (30%) and MS + 0.3 mg L-1 GA3 in the absence of light (30.5%). S. cineraria seeds were indifferent to light. Blue and green lights inhibited germination. Different concentrations of gibberellic acid (GA3) (0.1; 0.4; 0.6; 0.8; 1.0 and 2.0 mg L-1) and indole-3-acetic acid IAA (0.1; 0.3 and 1.0 mg L-1) were evaluated under white light and darkness. No concentration of GA3 enhanced seed germination percentage under white light. However, when the seeds were maintained in darkness, GA3 improved germination responses in all tested concentrations, except at 1.0 mg L-1. Under white light, these concentrations also increased the germination time and reduced germination rate. Germination rate, under light or darkness, was lower using IAA compared with GA3.

Exigências como matérias-primas vegetais fidedignas fazem parte das necessidades da fitoterapia moderna. A propagação in vitro permite a obtenção em massa de genótipos superiores, em curto espaço de tempo. Este estudo teve como objetivo desenvolver um protocolo de organogênese direta e indireta de Phyllanthus tenellus Roxb. Segmentos nodais de plântulas obtidas por germinação in vitro foram introduzidos em meio Murashige e Skoog adicionado de diferentes reguladores de crescimento vegetal: AIA (ácido indol-3-acético), AIB (ácido indol-3-butírico), AG3 (ácido giberélico-3) e CIN (cinetina). A maior taxa de proliferação foi obtida utilizando as combinações: AIB, CIN + AG3 (3,5 mg L-1) e AIB + CIN (2,4 mg L-1). A rizogênese foi intensificada após 40 dias de cultivo, atingindo 100% para todos os meios contendo AIB. A adição de ácido 2,4 diclorofenoxiacético (2,4D) proporcionou os melhores resultados para produção de calos friáveis. A aclimatização foi 100% eficaz para as plantas cultivadas em meio controle, com a diminuição da taxa de sobrevivência para as plantas cultivadas em meios adicionados de reguladores de crescimento.

Exigencies as ethic plant raw material are part of the needs of modern phytotherapy. Micropropagation offers opportunities to obtain mass propagation of superior genotypes in short time. This study aimed to develop a protocol of direct and indirect organogenesis of Phyllanthus tenellus Roxb. Nodal segments from plantlets obtained by in vitro germination were subcultured in modified Murashige and Skoog medium added with different plant growth regulators: IAA (indole-3-acetic acid), IBA (indole-3-butyric acid), GA3 (3-giberelic acid) and KIN (kinetin). The highest proliferation rate was obtained using the combinations: IBA, KIN + GA3 (3.5 mg L-1) and IBA + KIN (2.4 mg L-1). Rooting was intensified after 40 days, reaching 100% for all media with indole-3-butyric acid. Addition of 2,4 dichlorophenoxyacetic acid (2,4D) provided the best results for production of friable calli. Acclimatization was 100% effective for plantlets cultured in control medium, with decrease in survival rate in grown plantlets from media added with growth regulators.

Chemical composition of leaf volatiles of Rosmarinus officinalis and Baccharis dracunculifolia cultured in Southeast of Brazil has been characterized by GC/MS after simultaneous distillation-extraction. The main components in volatiles of these species showed in common α-pinene, myrcene, 1,8 cineole and camphor. Camphor was the major component among volatiles of B. dracunculifolia and R. officinalis with concentrations exceeding 25%. B. dracunculifolia volatiles possessed more sesquiterpenes (21.4%) than R. officinalis (16.7%), such as caryophyllene (1.9%) and α-humulene (0.4%). Lower concentrations of nerolidol and spathulenol were achieved in volatiles of B. dracunculifolia. Considering both species, there was a predominance of monoterpenes.

Atualmente, a hipertensão arterial é uma das maiores causas de morte e de doenças cardiovasculares. Os vasodilatadores são freqüentemente utilizados no tratamento da hipertensão. Extratos hidroalcoólicos de Alpinia purpurata de campo e de A. zerumbet cultivada in vitro sob diferentes reguladores de crescimento vegetal foram ensaiados no leito mesentérico de ratos Wistar. Os extratos de A. purpurata e A. zerumbet produziram efeito vasodilatador com padrão de resposta dose-dependente de duração prolongada. Extratos da espécie A. purpurata tiveram efeito vasodilatador de 87% na dose de 60 μg. O extrato obtido de folhas de A. zerumbet oriundas das culturas mantidas em meio contendo AIA (ácido indol acético) inibiu o relaxamento (17,4%) na dose de 90 μg em relação ao controle (MS0), com o qual foi verificado melhor efeito vasodilatador (60%). Estes resultados estão de acordo com a concentração de fenóis totais que foi 50% menor para os extratos de plantas cultivadas in vitro em AIA. A espécie A. purpurata foi pela primeira vez ensaiada quanto à atividade vasodilatadora. Os resultados obtidos indicaram a presença de substâncias fenólicas provavelmente correlacionadas à ação terapêutica de A. purpurata.

Nowadays, the high blood pressure is one of the main causes of death and cardiovascular diseases. Vasodilator drugs are frequently used to treat arterial hypertension. Experiments were undertaken to determine whether hydroalcoholic extracts obtained from leaves of field-grown Alpinia purpurata and A. zerumbet cultured in vitro under different plant growth regulators induce a vasodilator effect on Wistar rat mesenteric vascular bed pre-contracted with norepinephrine. Plant extracts were able to induce a long-lasting endothelium-dependent vasodilation. Efficiency on activity of A. purpurata reached 87% at concentration of 60 μg. The extract of A. zerumbet maintained in medium containing IAA, induced the relaxation (17.4%) at 90 μg, as compared to the control (MS0) that showed a better vasodilator effect (60%). These results are in agreement with the quantification of phenolic compounds in the extracts, which were 50% lower for those plants cultured in IAA. A. purpurata was assayed for the first time in relation to its vasodilator activity. This paper showed a strong probability of correlation between the pharmacological activities of A. purpurata with their content in phenolic compounds.