Growth, betacyanin production and bioactive potential of plants of the genus Alternanthera

The medicinal species of the genus Alternanthera with Alternanthera philoxeroides and Alternanthera tenella feature a variety of bioactive compounds, including flavonoids and betacyanins, pigments with antioxidant properties. Micropropagation can contribute to the study of bioactive plants it provides controlled growing conditions and permit the use of exogenous precursors or elicitors of the biosynthesis of substances of interest. Other promising tools in the study of medicinal plants are the tests to verify the bioactive potential of these species, using bioindicators. This work aimed to study the growth, production betacyanins and potential bioactive plant of the genus Alternanthera. In vitro experiments, nodal segments were inoculated on MS medium with different concentrations of tyrosine (0, 25, 50 and 75 μM). Plants were evaluated on the number of shoots and buds, height, root length, fresh weight of part shoots and roots and content betacyanin. In the species A. philoxeroides among the variables analyzed, only to shoot high, the highest concentration, showed a significant decrease. The root was affected from the first concentration occurring a decrease in the length of which the concentration of 75μM no formation of this organ. Betacyanin production in all treatments was higher in stem than leaf. For the testing of bioactive species A. philoxeroides extracts were obtained by maceration of dried leaves and stems, with subsequent liquid-liquid partition with solvents of increasing polarity (hexane, dichloromethane, ethyl acetate, butanol). Germination tests were performed with solutions of 0; 1,0; 2,0 e 3,0mg mL-1 with 30 seeds of lettuce and four replications. The antibacterial activity was evaluated by disc diffusion technique was tested where the sensitivity of five reference strains, with the extracts at a concentration of 100mg mL-1 .The ethyl acetate extract obtained from leaves at the highest concentration influenced only in the germination of lettuce, while the force was significantly affected by all extracts at concentrations of 1.0 mg mL-1. None of the extracts of A. philoxeroides the concentration tested interfered with the growth of the strains used. Variables in vitro growth of A. tenella showed a decrease in their average with increasing concentrations of tyrosine and the concentration of 75μM was toxic. However betacyanin production was enhanced with the addition of 50μM of the compound in the culture medium. These results demonstrate the positive effect of elicitation with the production of tyrosine betacyanin in both species, but applications in high concentrations are deleterious to plant growthAs espécies medicinais do gênero Alternanthera (Amaranthaceae), como Alternanthera philoxeroides e Alternanthera tenella apresentam uma variedade de compostos bioativos, entre eles os flavonoides e as betacianinas, pigmentos com propriedades antioxidantes. A micropropagação pode contribuir no estudo de plantas bioativas, pois proporciona condições de crescimento controladas e admitem o uso de precursores exógenos ou elicitores da biossíntese de substâncias de interesse. Outras promissoras ferramentas no estudo das plantas medicinais são os testes para verificação do seu potencial bioativo, utilizando para isso, organismos bioindicadores. Este trabalho teve o objetivo de estudar o crescimento, a produção de betacianinas e o potencial bioativo de plantas do gênero Alternanthera. Nos experimentos in vitro, segmentos nodais foram inoculados em meio MS com diferentes concentrações de tirosina (0; 25; 50 e 75μM). As plantas foram avaliadas quanto ao número de brotos e gemas, altura, comprimento das raízes, massa fresca da parte aérea e das raízes e teor de betacianina. Na espécie A. philoxeroides dentre as variáveis de parte aérea analisadas somente a altura, na maior concentração, apresentou um decréscimo significativo. A raiz foi afetada a partir da primeira concentração ocorrendo um decréscimo no comprimento sendo que na concentração de 75μM não houve formação desse órgão. A produção de betacianina, em todos os tratamentos, foi maior no caule do que na folha. Para os ensaios de atividade bioativa da espécie A. philoxeroides, os extratos foram obtidos por meio da maceração, de folhas e caule secos, com posterior partição líquido-líquido, com solventes de polaridade crescentes (hexano, diclorometano, acetato de etila, butanol). Os testes de germinação foram realizados com as soluções nas concentrações de 0; 1,0; 2,0 e 3,0mg mL-1, com 30 sementes de alface e quatro repetições. A atividade antibacteriana foi avaliada por técnica de difusão de discos onde foi testado a sensibilidade de cinco cepas de referência, com os extratos na concentração de 100mg mL-1. O extrato acetato de etila obtido de folhas na maior concentração influenciou somente na germinabilidade da alface, enquanto o vigor foi afetado significativamente por todos os extratos a partir da concentração de 1,0mg mL-1. Nenhum dos extratos de A. philoxeroides na concentração testada interferiu no crescimento das cepas utilizadas. As variáveis de crescimento in vitro de A. tenella mostraram diminuição nas suas médias com o aumento das concentrações de tirosina sendo que a concentração de 75μM foi tóxica. Contudo a produção de betacianina foi beneficiada com a adição de 50μM deste composto no meio de cultura. Estes resultados demonstram o efeito positivo da elicitação com tirosina na produção de betacianina, nas duas espécies, porém aplicações em altas concentrações são deletérias para o crescimento das planta

Potential allelopathic and antibacterial activity from Alternanthera philoxeroides

ABSTRACT Alternanthera philoxeroides is a promising species in the search for new molecules with biological effects. To accelerate this bioprospection process of A. philoxeroides, the chosen methods (allelopathy and discdiffusion technique) are excellent alternatives. The purpose of this study was to evaluate the allelopathic effects and verify the antibacterial properties of extracts from A. philoxeroides. To evaluate the antibacterial activity, disc diffusion technique was used with five reference strains. The germination test was conducted with the extracts at concentrations of 0, 1.0, 2.0 e 3.0 mg mL-1 in a petri dish with 30 lettuce seeds and for the growth trial 10 seeds were used. The experimental design was completely randomized in factorial 4 × 2 × 4 (four extracts, two sources, four concentrations), with four replicates.The results indicate that the lettuce germination was influenced by ethyl acetate fraction obtained from leaves of A. philoxeroides at the highest concentrations; however, the germination speed and the radicle growth were significantly affected by all the extracts starting at 1mg mL-1 concentration. Those results demonstrate allopathic effect of different extracts of this plant, although they have not interfered with the strains growth used in this experiment

Production of pigments in Alternanthera sessilis calli mediated by plant growth regulators and light

Among the compounds produced by plants, pigments such as betalains have received attention from both food and pharmaceuticals industries. The Alternanthera sessilis species produces these pigments, though in small quantities, and so it is necessary to increase production. Thus, many studies use elicitors that are capable of triggering physiological or morphological responses in plants. The objective was to establish callus production in A. sessilis grown under different combinations of growth regulators and light qualities and to assess whether these factors can increase betalain and flavonoid production. Leaf and internodal explants in MS medium with different growth regulators were used to obtain calli, which were subsequently transferred to a betacyanin induction medium remaining for 40 days under different light qualities (white, blue, red, and dark). The most suitable treatment for callus formation and subsequent betalain and flavonoid induction was to combine a medium containing 6.7 µmol L-1 2,4-D and 9.0 µmol L-1 BAP and blue light. Physical elicitation by light combined with appropriate concentration of growth regulators on calli can increase production of commercially important metabolites.

Callus induction and betacyanin quantification by HPLC/MS-MS in Alternanthera brasiliana (L.) Kuntze

ABSTRACT The objective of this study was to establish a protocol for callus induction and betacyanin production in plants of Alternanthera brasiliana. Explants of A. brasiliana and five combinations of cytokinin and auxin were used for callus induction. Calli were transferred to a Betacyanin Induction Medium (MIB), composed of MS, with 0.5 mg L-1 of thidiazuron (TDZ) and 1 mg L-1 of naphthaleneacetic acid (NAA), and kept in the light for 45 days. The aspect and intensity of pigments were assessed and total betacyanins were quantified in high performance liquid chromatography (HPLC). The combination of internodal segments and a medium containing equilibrium concentrations of auxins and cytokinins was the most efficient metod to induce calli and increased production of betacyanins. The presence of amaranthine in calli of A. brasiliana justifies its medical use and the consequent need for future studies for the large-scale production of this molecule

Produção de betacianina em erva-de-jacaré cultivada in vitro com diferentes concentrações de sulfato de cobre Betacyanin production in alligator weed, grown in vitro, with different concentrations of copper sulfate

A manipulação da concentração dos microelementos no meio de cultura representa uma boa estratégia para aumentar a produção de metabólitos secundários em cultura de tecidos. Este trabalho teve por objetivo demonstrar a influência do sulfato de cobre na produção de betacianina e na multiplicação de plantas de Alternanthera philoxeroides (erva-de-jacaré) cultivadas in vitro. Segmentos nodais, obtidos de plantas pré-estabelecidas e após três subcultivos, foram inoculados em meio MS, com diferentes concentrações de CuSO4 (0; 25; 75; 125; 175; 200µM). O delineamento experimental utilizado foi inteiramente casualizado, com seis concentrações de CuSO4 e cinco repetições representadas por um frasco contendo quatro explantes. Os resultados foram submetidos à análise de variância, à regressão polinomial e a análises de correlação entre as variáveis morfológicas e a produção de betacianina. Concentrações acima de 75µM diminuíram a altura das plantas, o número de brotos e gemas e o crescimento radicular, enquanto que, na concentração de 125µM, houve a maior produção de massa fresca. A produção de betacianina aumentou 60% em relação ao controle com 175µM de CuSO4. Ocorreu crescimento das plantas em todas as concentrações de CuSO4 testadas, com exceção de 200µM, sendo esta considerada tóxica.The manipulation of concentration of trace elements in the culture medium represents a good strategy to increase the production of secondary metabolites in tissue culture. This study aimed to demonstrate the influence of copper sulphate in the production of betacyanin and the multiplication of plants of Alternanthera philoxeroides (alligator weed) cultured in vitro. Nodal segments, obtained from plants pre-stablished and after three subcultures, were inoculated in basic MS with different concentrations of CuSO4 (0, 25, 75, 125, 175, 200µM). The results were subjected to analysis of variance, polynomial regression and correlation analysis between the morphological variables and the production of betacyanin. The experimental design was completely randomized with six concentration of CuSO4 and five replicates represented by a bottle containing four explants. Concentrations above 75µM decreased the height of the plant, number of shoots and buds and root growth, while the concentration of 125µM resulted in the highest fresh mass production. Betacyanin production increased 60% over control with 175µM CuSO4. Plant growth occurred in all tested concentrations of CuSO4, with the exception of 200µM which was toxic