Cultura de tecidos e análise fitoquímica de Arachis repens HANDRO.

Os representantes do gênero Arachis são conhecidos como amendoim, e suas sementes possuem grande quantidade de óleos e proteínas, podendo ser consumidas cruas. A espécie mais cultivada do gênero é Arachis hypogaea, que possui grande importância comercial. Outras espécies são também utilizadas para controle das ervas daninhas e cobertura do solo, assim como ornamentais e na alimentação. Arachis repens, popularmente conhecida como grama amendoim, é utilizada como planta ornamental, na formação de pastagens, como forragem e para cobertura do solo em substituição a várias espécies comuns de grama. Diversas substâncias bioativas têm sido identificadas em A. hypogaea. Contudo, estas substâncias ainda não foram descritas em outras espécies do gênero. O objetivo deste trabalho foi o estabelecimento de sistemas de cultivo in vitro para Arachis repens, visando à micropropagação e à análise comparativa da produção de resveratrol em extratos alcoólicos dos materiais produzidos in vitro, em comparação com as plantas in vivo. Segmentos nodais, internodais e foliares foram excisados de plantas in vitro e inoculados em meio MS suplementado com diferentes concentrações de BAP, TDZ, AIA e KIN, utilizados isoladamente ou em combinação. Os explantes apresentaram respostas distintas de acordo com o regulador de crescimento utilizado. Na presença de BAP foi observada a formação de calos compactos, com formação de brotos em diferentes taxas. Na presença de TDZ, segmentos nodais e internodais formaram calos compactos em todas as concentrações. Segmentos nodais cultivados na presença de diferentes concentrações de AIA em combinação com KIN apresentaram a formação de calogênese. Foi também realizada uma avaliação da atividade antioxidante e a presença de fenóis totais em extratos de diferentes materiais in vivo e produzidos in vitro. Nas análises por meio de CLAE, foi possível detectar a presença de resveratrol tanto nos materiais in vivo, quanto os produzidos in vitro. Dessa forma, o trabalho forneceu resultados preliminares sobre as possibilidades de utilização de A. repens para a produção de metabólitos especiais.

In vitro morphogenesis and cell suspension culture establishment in Piper solmsianum C. DC. (Piperaceae)

In vitro morphogenesis and cell suspension culture establishment in Piper solmsianum C. DC. (Piperaceae)). Piper solmsianum is a shrub from Southeast Brazil in which many biologically active compounds were identified. The aim of this work was to establish a cell suspension culture system for this species. With this in mind, petiole and leaf explants obtained from in vitro plantlets were cultured in the presence of different plant growth regulator combinations (IAA, NAA, 2,4-D and BA). Root and indirect shoot adventitious formation, detected by histological analysis, was observed. Besides the different combinations of plant growth regulators, light regime and the supplement of activated charcoal (1.5 mg.l(-1)) were tested for callus induction and growth. Cultures maintained in light, on a 0.2 mg.l(-1) 2,4-D and 2 mg.l(-1) BA supplemented medium, and in the absence of activated charcoal, showed the highest calli fresh matter increment. From a callus culture, cell suspension cultures were established and their growth and metabolite accumulation studied. The achieved results may be useful for further characterization of the activated secondary metabolites pathways in in vitro systems of P. solmsianum.

Cultura de embrião e indução de brotos in vitro para micropropagação do pinhão-manso

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

Induction and Morpho-Ultrastructural Analysis of Organogenic Calli of a Wild Passionfruit

This work studied a new protocol for organogenic calli induction and characterization of the morphology and ultrastructure of callogenesis in leaf explants of Passiflora gibertii N. E. Brown, a native passion fruit species from Brazil. Calli induction was performed in different growth conditions (light and dark), different MS medium salt concentrations (MS and MS half strength) and the presence or absence of coconut water. The leaf explants maintained in the dark were more responsive to bud formation. In order to reduce spending on in vitro culture, the most suitable induction medium for P. gibertii organogenesis could, therefore be the MS half strength salt concentration medium maintained in the dark. The addition of coconut water to the culture medium was essential for both calli induction and bud formation. The morphological and ultrastructural features of the organogenic calli were isodiametric cells, characterized by an organized cellular system, nucleus with prominent nucleoli, presence of starch grains and dense cytoplasm rich in endoplasmic reticulum. The scanning electron microscopy demonstrated that buds were present on these calli.

Somatic embryogenesis from ovaries of sweet orange cv. Tobias

Callogenesis, somatic embryogenesis, and regeneration were obtained from tissues of unfertilized ovaries of sweet orange (Citrus sinensis Osbeck.) cv. Tobias. The influence of two modified basal media, woody plant medium (WPM) and N6 medium, to induce callus formation from pistils was determined. Overall, high frequencies of callogenesis were observed when either medium was used. However, initial culture of explants in WPM medium followed by transfer of callus to N6 medium resulted in higher frequency of callus induction (of 2.30 callus per explant that were larger than 0.5 cm in size), and of subsequent development of embryogenic callus (10%). A total of 125 somatic embryos were obtained. After 6 months of culture, 72% of somatic embryos germinated into plantlets. These plantlets were subsequently micrografted in vitro, and then acclimatized. Ploidy of these plants were determined using flow cytometry and TRAPS molecular markers were used to confirm their maternal origin.

In vitro organogenesis of Eucalyptus grandis: effects of boron and calcium

The in vitro organogenesis of woody species plays an essential role in the improvement of forest products by providing saplings with high commercial value. Furthermore, mineral nutrition plays an important role in the induction of organogenic responses. The objective of this study was to evaluate the effects of boron and calcium in the organogenesis of nodal segments from seedlings of Eucalyptus grandis growing under in vitro conditions. The concentration of boron and calcium in MS medium was modified to induce organogenic responses in 45-day-old nodal segments used as explants. After 60 days, the fresh weight, dry weight, ratio of fresh and dry weight, relative water content and relative matter content accumulated by the explants were evaluated. The concentrations of boron and calcium in the culture medium influenced the in vitro organogenic control of Eucalyptus grandis. Reduced combinations of boron and calcium induced callus formation and dry matter accumulation in the explants. A boron concentration of 100% (1.10 mg L-1) combined with 100% (119.950 mg L-1) and 200% (239.900 mg L-1) of calcium, and 200% (2.20 mg L-1) of boron combined with 100% (119.950 mg L-1) of calcium allowed the induction of well-developed buds, which can be used for the regeneration of micro-plants.

Dedifferentiation of Leaf Cells and Growth Pattern of Calluses of Capsicum annuumcv. Etna.

Background:In vitrocell suspension cultivation systems have been largely reported assafe and standardized methods for production of secondary metabolites with medicinaland agricultural interest.Capsicum annuumis one of the most widely grown vegetablein the world and its biological activities have been demonstrated against insects, fungi,bacteria and other groups of organisms. The determination of procedures for thededifferentiation of cells into callus cells and the subsequent study of the callus growthpattern are necessary for the establishment of cellsuspensions and also to subsidizestudies regarding the bioactivity of its secondarymetabolites. To date, no study hasdescribed the development of protocols for callus induction inC. annuumL. cv. Etna. Objective:The objective of this study was to establish a protocol for dedifferentiationof leaf cells of the cultivarC. annuumcv. Etna and to determine the growth pattern ofthe calluses with a focus on the deceleration phase, when the callus cells must besubcultured into a liquid medium in order to establish cell suspension cultivationsaiming at the production of secondary metabolites.Results:The treatment that resultedin the highest %CI, ACCC and callus weight was thecombination of 4.52 μ M 2,4-D +0.44 μ M BA. The calluses produced were friable andwhitish and their growth patternfollowed a sigmoid shape. The deceleration phase started on the 23rdday of cultivation.Conclusion:Callus induction in leaf explants ofC. annuumcv. Etnacan be achieved inMS medium supplemented with 4.52 μ M 2,4-D + 0.44 μ MBA, which results in highcellular proliferation; in order to start a cell suspension culture, callus cells on the 23rdday of culture should be used.