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O artigo não contém resumo em português. The use of cell and plant tissues culture techniques to produce economically important active metabolites has been growing. Among these substances, azadirachtin (AZA), produced by the neem tree (Azadirachta indica), has received considerable attention due to its bioinsecticide action. The main goal of this work was to analyze the AZA levels in neem cotyledonary calli. The calli were grown in agitated Woody Plant Medium (WPM) liquid medium, supplemented with glucose (Gl), hydrolyzed casein (HC) and methyl jasmonate (MeJ) as elicitor agent. An interaction was observed between these substances, depending on in vitro cultivation time with orbital agitation. The highest concentrations (average of 0.2470 µg g-1) of AZA were produced in the first and second weeks of culture when the cell mass was grown in a medium with 2% Gl v/v, 500 mg L-1 HC and 100 µM of MeJ. This corresponded to approximately 57% of the AZA content stored in the donor plants seeds, used as a source of explants to induce in vitro callus formation. It was concluded that the nutrition, as well as the concentration of MeJ as signal transduction of secondary metabolism in neem cells, might influence the AZA content produced in vitro.

A propagação in vitro tem se destacado como uma técnica efetiva na produção em larga escala de plantas sadias de morangueiro. Neste trabalho estudamos a hiperhidricidade associado a capacidade de multiplicação in vitro na propagação de duas variedades de morangueiro (Fragaria x ananassa Duch. "Dover" e "Burkley"). Plantas mantidas em meio de cultura MS, suplementadas com 1.0 mg L-1 de BA fora individualizadas e transferidas para o mesmo meio com Ágar (6.5 g L-1) ou Phytagel® (2,5 g L-1) e BA em diferentes concentrações (0; 0,5; 1,0; 2,0 e 3,0 mg L-1). Foram realizadas análises bioquímica e anatômicas, além da caracterização morfológica do material hiper-hídrico. A análise dos dados mostrou: a) o incremento na concentração de citocinina aumentou a freqüência de hiper-hidricidade para ambas as variedades; b) concentrações maiores que 2,0 mg L-1 de BA, com a substituição do Ágar pelo Phytagel® induziu a maior formação de ramos hiper-hídricos; e c) a adição de BA induziu o estresse oxidativo, caracterizado pelo incremento da atividade antioxidante e peroxidação de lipídeos, bem como alterações a nível celular, como má formação dos estômatos e células epidérmicas. Concluindo, o meio de cultura contendo 0,5 mg L-1 de BA solidificado com Ágar promoveu menor porcentagem de hiper-hidricidade associado com maiores taxas de proliferação de gemas em morangueiro. In vitro propagation has become an effective practice for large-scale production of strawberry plants. The objective of this study was to evaluate the hyperhydricity and the multiplication capacity of two strawberry varieties (Fragaria x ananassa Duch. 'Dover' and 'Burkley') propagated in vitro. Plants maintained in MS medium supplemented with 1.0 mg L-1 BA were individualized and transferred to the same medium solidified with Agar (6.5 g L-1) or Phytagel® (2.5 g L-1) and BA at different concentrations (0; 0.5; 1.0; 2.0 and 3.0 mg L-1). Biochemical and anatomical analyses were carried out, as well as the analysis of the morphological hyperhydricity characteristics. The analysis of data showed: a) the increase in cytokinin concentration increased hyperhydricity frequency in both varieties; b) at concentrations up to 2.0 mg L-1 BA, the replacement of Agar by Phytagel® induced a higher formation of hyperhydric shoots; and c) the addition of BA induced oxidative stress, which is characterized by increased antioxidant activity and lipid peroxidation, as well as alterations at the cellular level, such as malformation of stomata and epidermal cells. In conclusion, the culture medium containing 0.5 mg L-1 BA solidified with Agar provided lower hyperhydricity percentages in association with higher rates of shoot proliferation in strawberry.

The hypothesis that application of exogenous glycine betaine (GBEX) may attenuate the effects of mild water deficit in leaf gas exchange and lipid peroxidation in Carapa guianensis was examined. For this reason, 110-d old plants were sprayed with 0, 25, and 50 mM GBEX and then subjected to two watering regimes. In the first, irrigation was continuously performed to maintain the soil near to field capacity (watered plants). In the second, irrigation was withheld and water deficit resulted from progressive evapotranspiration (water-stressed plants). Treatment comparisons were assessed when predawn leaflet water potential (Ψpd) of stressed plants reached −1.28 ± 0.34 MPa. Regardless of the watering regime, significant (P

Young Carapa guianensis plants were examined under well-watered (control) and water-deficit conditions with the aim to evaluate possible relationship between diurnal changes in leaflet gas exchange with lipid peroxidation and adjustments in antioxidative responses. Treatment comparisons were assessed when leaflet water potential (Ψw) in water-stressed plants reached around −2.5 ± 0.5 MPa at pre-dawn. Regardless of watering regime, the highest net CO2 assimilation rate and stomatal conductance were recorded until 9:00 h. Control plants showed diurnal increases in transpiration, while it was strongly decreased in water-stressed plants. Diurnal decreases in intercellular to ambient CO2 concentration ratio were just observed in stressed plants. Regardless of watering regime, non-significant changes (P > 0.05) in Ψw and relative water content were registered throughout the day; however, both variables were significantly lower (P

Changes in chloroplastidic pigments, gas exchange and carbohydrate concentrations were assessed during the rapid initial expansion of C. guianensis leaflet. Leaves at metaphyll stage were tagged and assessments were carried out 14, 17, 20, 23, 27, and 31 days later. Pigments synthesis, distribution and accumulation were uniform among leaflet sections (basal, median and apical). Chlorophyll (Chl) a, Chl b, Chl (a+b), and total carotenoids (Car) concentrations were significantly increased after 27 days from metaphyll, and the most expressive increases were parallel to lower specific leaflet area. Chl a/b was lower on day 14 and it was increased on subsequent days. Negative net photosynthesis rate (P N), and the lowest stomatal conductance (g s) and transpiration (E) were registered on day 14, following significant increases on subsequent days. The Chl (a+b) and Chl a effects on P N were more expressive until day 20. Intercellular to ambient CO2 concentration ratio (C i/C a) was higher on day 14 and lower on subsequent days, and no stomatal limitation to CO2 influx inside leaflets was observed. Leaflet temperature was almost constant (ca. 35°C) during leaflet development. Sucrose and starch concentrations were increased in parallel to increases in P N. Altogether, these results highlight the main physiological changes during C. guianensis leaflet expansion and they should be considered in future experiments focusing on factors affecting P N in this species.

In vitro propagation of neem (Azadirachta indica A. Juss.) may offer an efficient alternative to seed propagation of this species. For optimization of in vitro propagation, different basal salt formulations, growth regulators, and culture container sealants (polytetrafluoroethylene hydrophobic membranes [PTFE]) were evaluated. Nodal segments cultured on Murashige and Skoog (MS) medium showed the highest shoot formation per explant (1.67). Explants cultured in flasks containing MS medium with 0.5 mg L−1 benzyladenine, 0.5 mg L−1 kinetin, and 0.05 mg L−1 naphthaleneacetic acid, and sealed with two PTFE membranes, produced the highest number of shoots (4.04). In contrast, explants cultured in flasks without membranes showed leaf chlorosis and senescence. For plant recovery, regenerants were acclimatized in a substrate of coconut fiber and eucalyptus bark (1:1) and showed 80% survival. Our results indicated that the use of flasks with vents was beneficial for in vitro propagation of this important plant.

The ecdysteroid 20-hydroxyecdysone (20E) is a steroid hormone found in insects and few plants. To analyze its temporal and spatial distribution in Pfaffia glomerata (Amaranthaceae), we selected among 71 accessions, the accession 13 which showed the highest root 20E total quantity (1.48 g/plant root). The 20E was constantly detected in flowers, leaves, stems, and roots, but its percentage was variable throughout its development. The highest 20E percentage was found in flowers (0.82%), roots (0.66%), leaves (0.60%), and stems (0.24%). While stems showed the least variable 20E percentage (0.13% - 0.24%), followed by roots (0.42% - 0.66%), and flowers (0.47% - 0.82%). Leaves showed the greatest 20E percentage variation (0.21% - 0.60%). The stem and leaves 20E total amount remained constant, showing a little variation while root 20E total amount increased over time, suggesting that 20E may be accumulated in roots. These findings suggest that 20E has tissue specific functions in plant and may have ecological significance since uncoordinated pulses of 20E are often lethal to insects.