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- How to monitor the acclimatization of micropropagated plants - From in vitro to the field?Publication . Osório, Maria Leonor; Gonçalves, Sandra; Coelho, N.; Romano, Anabela; Osório, JúlioThe high mortality experienced by plants during ex vitro transplantation stage is the major bottleneck in large scale application of micropropagation. The abnormal physiological and anatomical characteristics of micropropagated plantlets require that they should be gradually acclimatized to the environment of the greenhouse or field. Environmental factors, especially relative humidity and irradiance, play a critical role in both physiological and biochemical functions, namely water relations, gas exchange, photosynthetic efficiency and water oxidation, during transition from in vitro to ex vitro conditions. Throughout this period of transition and acclimatization substantial changes in those characteristics are necessary to achieve a successful micropropagation. This adaptation should be accurately monitored using relevant physiological parameters including pigment content, chlorophyll a fluorescence imaging, net photosynthetic rate, transpiration rate, and stomatal conductance. Growth traits and oxidative stress markers, such as electrolyte leakage, lipid peroxidation and hydrogen peroxide content, should be also analyzed. These parameters must be assessed during the course of ex vitro acclimatization in several periods and compared with those of plants growing in their natural habitat. The performance plants of Tuberaria major, an endangered species endemic from the Algarve region (Portugal), during their acclimatization through indoor-to-outdoor conditions was monitored using several physiological and biochemical traits as indicators. In view of the results, we concluded that the transplantation protocol described can be used for restoration purposes, contributing to the preservation of the species. © ISHS 2013.
- Differences in Al tolerance between Plantago algarbiensis and P. almogravensis reflect their ability to respond to oxidative stressPublication . Martins, Neusa; Osório, Maria Leonor; Gonçalves, Sandra; Osório, Júlio; Romano, AnabelaWe evaluated the impact of low pH and aluminum (Al) on the leaves and roots of Plantago almogravensis Franco and Plantago algarbiensis Samp., focusing on energy partitioning in photosystem II, H2O2 levels, lipid peroxidation, electrolyte leakage (EL), protein oxidation, total soluble protein content and antioxidant enzyme activities. In both species, Al triggered more changes in oxidative metabolism than low pH alone, particularly in the roots. We found that Al increased the levels of H2O2 in P. algarbiensis roots, but reduced the levels of H2O2 in P. almogravensis leaves and roots. Neither low pH nor Al affected the spatial heterogeneity of chlorophyll fluorescence, the maximum photochemical efficiency of PSII (F-v/F-m), the actual quantum efficiency of PSII (I center dot(PSII)) or the quantum yields of regulated (I center dot(NPQ)) and nonregulated (I center dot(NO)) energy dissipation, and there was no significant change in total soluble protein content and EL. In P. algarbiensis, Al increased the carbonyl content and the activities of superoxide dismutase (SOD) and catalase (CAT) in the roots, and also CAT, ascorbate peroxidase and guaiacol peroxidase activities in the leaves. In P. almogravensis, Al reduced the level of malondialdehyde in the roots as well as SOD activity in the leaves and roots. We found that P. almogravensis plantlets could manage the oxidative stress caused by low pH and Al, whereas the P. algarbiensis antioxidant system was unable to suppress Al toxicity completely, leading to the accumulation of H2O2 and consequential protein oxidation in the roots.
- Influence of transient shade periods on the effects of drought on photosynthesis, carbohydrate accumulation and lipid peroxidation in sunflower leavesPublication . Correia, Maria João; Osório, Maria Leonor; Osório, Júlio; Barrote, Isabel; Martins, Mónica; David, Maria ManuelaThe effects of a slow-imposing two-weeks soil drying period, and subsequent re-watering, on leaf water potential (Ψ), gas exchange rates, chlorophyll fluorescence and on the concentrations of malondialdehyde (MDA) and non-structural carbohydrates (starch, hexose and sucrose) were determined in mature leaves of sunflower plants growing under controlled environmental conditions. To assess how transient shade periods, associated with increased cloud cover, may influence drought-induced effects on carbon assimilation, measurements were carried out both in plants kept under the photosynthetic photon flux density (PPFD) prevailing during the growth period and stress imposition (750 μmol m-2 s-1), and in plants subjected to a 5 hours-long period under a lower PPFD (200 μmol m-2 s-1). In plants kept under high PPFD, Ψ, stomatal conductance (g), net CO2 uptake rate (A), the quantum yield of photosystem II electron transport (Φe), the photochemical efficiency of open PSII reaction centres (Fv’/Fm’) and * Corresponding author. Tel: +351 289800932; fax: +351 289818419. E-mail address: jcorreia@ualg.pt (M.J. Correia) 1 the diurnal accumulation of total non-structural carbohydrates (TNC) were significantly depressed at the end of the soil drying period, whereas non-photochemical quenching (NPQ), the concentrations of MDA and the predawn pools of soluble sugars were found to increase. Under high-light level, drought-induced effects on lipid peroxidation, chlorophyll fluorescence parameters and gas exchange rates were fully reversed upon re-watering. However, the inhibition of diurnal accumulation of TNC still persisted two days following re-watering, suggesting that carbohydrate export rates were enhanced following stress relief. An overall positive effect was found upon transferring water-stressed plants to low light level, as indicated by the increases in Ψ, intrinsic water use efficiency (A/g), Φe and Fv’/Fm’, as well as the reversal of drought-induced enhancement of both NPQ and MDA concentration. Despite g being similar in shaded well-watered and re-watered plants, the latter exhibited net CO2 uptake rates below those found in well-watered leaves under the same light conditions, together with a diurnal decrease in the concentration of TNC that was mainly attributable to the depletion of starch and sucrose. These results indicate that contrasting with the positive effects of shading on water-stressed plants, low PPFD may negatively affect the recovery of net photosynthesis following stress relief.
- Influence of enhanced temperature on photosynthesis, photooxidative damage, and antioxidant strategies in Ceratonia siliqua L. seedlings subjected to water deficit and rewateringPublication . Osório, Maria Leonor; Osório, Júlio; Vieira, Ana; Gonçalves, Sandra; Romano, AnabelaPredicted future climatic changes for the Mediterranean region give additional importance to the study of photooxidative stress in local economic species subjected to combined drought and high-temperature conditions. Under this context, the impact of these stresses on photosynthesis, energy partitioning, and membrane lipids, as well as the potential ability to attenuate oxidative damage, were investigated in Ceratonia siliqua L. Two thermal regimes (LT: 25/18ºC; HT: 32/21ºC) and three soil water conditions (control, water stress, and rewetting) were considered. HT exacerbated the adverse effects of water shortage on photosynthetic rates (PN) and PSII function. The decrease in PN was 33% at LT whereas at HT it was 84%. In spite of this, the electron transport rate (ETR) was not affected, which points to an increased allocation of reductants to sinks other than CO2 assimilation. Under LT conditions, water stress had no significant effects on yield of PSII photochemistry (ΦPSII) and yields of regulated (ΦNPQ) and nonregulated (ΦNO) energy dissipation. Conversely, drought induced a significant decrease of ΦPSII and a concomitant increase of ΦNO in HT plants, thereby favouring the overproduction of reactive oxygen species (ROS). Moreover, signs of lipid peroxidation damage were detected in HT plants, in which drought caused an increase of 40% in malondialdehyde (MDA) content. Concurrently, a marked increase in proline content was observed, while the activities of catalase (CAT) and ascorbate peroxidase (APX) were unaffected. Despite the generation of a moderate oxidative stress response, C. siliqua revealed a great capability for photosynthetic recovery 36 h after rewatering, which suggests that the species can cope with predicted climate change.