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- Influence of nutrient availability on drought-induced changes in the activity of antioxidant enzymes in sunflower leavesPublication . David, Maria Manuela; Barrote, Isabel; Correia, Maria JoãoWe aimed to evaluate if plants with different nutrient availability evidenced a different capacity to overcome drought-induced oxidative stress. Membrane peroxidative damages as MDA concentration and the activity of several antioxidant enzymes were determined in leaves of well watered (WW) and water stressed (WS) H. annuus plants grown either with adequate (Adeq Nutr) or limited (Limit Nutr) nutrient regimes. Constitutive capacity to eliminate ROS was not overall changed by growing plants with different nutrient supply regimes, but a diverse enzyme-dependent response was observed. In response to drought SOD and DHAR activity increased only in plants with limited nutrient supply, while in plants with adequate nutrient supply the activity of these enzymes did not change and were constitutively higher, but the activity APX increased by 50%. The subtle observed changes in the activity of the antioxidant enzymes are discussed, given that no oxidative damage was observed.
- The effects of drought on net CO2 uptake, chlorophyll fluorescence and photosynthetic pigments in mature and young leaves of sunflowerPublication . Osório, Maria Leonor; Barrote, Isabel; Osório, Júlio; David, Maria Manuela; Correia, Maria JoãoPhotosynthesis, photosystem II activity and photosynthetic pigments were analysed in mature and Young leaves of sunflower (Helianthus annuus L.) during dehydration and rehydration, in order to determine the influence of leaf age on the sensitivity to drought. Dehydration caused a more pronounced inhibition in photosynthetic rates in mature leaves compared to younger leaves, but 24 h after rewatering the recovery was complete in spite of leaf age. After exposure to drought, mature leaves showed a larger reduction in the actual PSII photochemical efficiency and a higher increase in nonradiative energy dissipation than young leaves. In well-watered plants the de-epoxidation level represented by the ratio A+Z/V+A+Z was higher in young leaves. However drought had opposite effects on both type of leaves: in young leaves A+Z/V+A+Z decreased with water deficit, while in mature leaves increased with soil drying. The results obtained suggest that leaf age influence the “strategy” to avoid irreversible damage of PSII activity under drought and that older leaves are more drought-sensitive than younger leaves of sunflower.
- Effects of water deficit on the activity of nitrate reductase and content of sugars, nitrate and free amino acids in the leaves and roots of sunflower and white lupin plants growing under two nutrient supply regimesPublication . Correia, Maria João; Fonseca, Filomena; Azedo-Silva, João; Dias, Celeste; David, Maria Manuela; Barrote, Isabel; Osório, Maria Leonor; Osório, JúlioThe effects of a two weeks soil drying period on the activity of nitrate reductase (NR; EC 1.6.6.6) were studied on Helianthus annuus L. and non-nodulated Lupinus albus L. plants, growing under two nutrient supply regimes. NR activity was assessed in leaf and root extracts, by measuring the activity of the unphosphorylated active form (NRact), the maximal extractable activity (NRmax) and the activation state. To get insight into potential signalling compounds, nitrate, amino acids and soluble sugars concentrations were also quantified. On both species, foliar NRact and NRmax were negatively affected by soil drying and reduced supply of nutrients, the observed changes in NR activity being linearly-correlated with the depletion of nitrate. Similar results were obtained in the roots of sunflower. Conversely, in white lupin roots NRmax was found to be independent of tissue nitrate concentration. Regardless of the species and organ, the activation state of the enzyme was unaffected by the nutrient supply regime. In well-watered sunflower roots only about 50% of the existing NR was unphosphorylated, but the activation state increased significantly in response to drought. In contrast, lupin roots always exhibited NR activation state values close to 80% or even higher. At the leaf level, NR activation state was hardly changed in response to soil drying. The contribution of changes in the concentrations of soluble 5 sugars and amino acids to explain the observed variations in NR activity are discussed.
- Alterações sazonais da fotossíntese e do ciclo das xantófilas em alfarrobeira (Ceratonia siliqua L.)Publication . Barrote, Isabel; Correia, Maria João; David, Maria ManuelaThis study aimed to follow the seasonal changes in the photosynthetic performance of two Ceratonia siliqua cultivars – Mulata and Mulata do Espargal – growing in a non-irrigated orchard in Algarve (South Portugal). Measurements and sampling (leaf water potential, leaf gas exchange, pigments and dark-adapted chlorophyll fluorescence) were taken in days with similar photon flux density, during the dry season in June and July, and in November after the first autumnal rainfalls (»91 mm) when minimum temperatures droped below 10°C. Photosynthetic rates remained low for both cvs, and did not recover in November despite the observed increase in stomatal conductance and y. Fv/Fm slowly decreased from June to November, specially at midday. This changes in Fv/Fm were attributed mainly to increasing F0 which might reflect structural changes or damages at the chloroplast membranes level. Dry season conditions seemed to have triggered some thermal photoprotective mechanisms, as indicated through the increased ratio A+Z/VAZ. In November, however, the ratio A+Z/VAZ decreased. We suggest that the lack of recovery of A associated with higher F0 measured in both cvs, either at pre-dawn or midday, could be attributed to structural changes within the membranes mediated either by rehydration and/or by chilling temperatures.
- Leaf age effects on photosynthetic activity and sugar accumulation in droughted and rewatered Lupinus albus L. plantsPublication . David, Maria Manuela; Coelho, Dulce; Barrote, Isabel; Correia, Maria JoãoChanges in the photosynthetic rate (A), stomatal conductance (g), water relations, photosynthetic pigments, Rubisco and soluble sugars accumulation were studied in different aged leaves of white lupin during soil drying and following rehydration. In water-stressed plants, A and g sharply declined and recovered only partially after rewatering. The way Ci and A/gchanged with drought was strongly dependent on leaf age; only in the young leaves did A/g increase and Ci decrease. Drought induced accumulation of soluble sugars was also age dependent, decreasing as leaves aged. In response to soil drying, the contents of photosynthetic pigments, total soluble protein and Rubisco protein increased in the young leaves and were either not affected or slightly decreased in the older ones. Rehydration accentuated the losses in pigments and Rubisco in the old leaves of water-stressed plants. These results suggest that the contribution of mesophyll limitations to explain drought inhibition of photosynthesis increases with leaf age, decreasing the ability to recover after rewatering. In young leaves the tolerance of the photosynthetic apparatus to dehydration and rehydration episodes is high and it is associated with high contents of Rubisco and in soluble sugars, particularly hexoses.
- Photosynthetic responses of Lupinus albus to soil water fluctuationsPublication . Barrote, Isabel; Osório, Maria Leonor; Osório, Júlio; David, Maria Manuela; Correia, Maria JoãoLupinus albus plants were grown in 3 dm3 pots in a semi-controled greenhouse. Two water regimes were imposed: water deficit (S, 47% of soil capacity), and control (T, 86% of soil capacity). Plants water status was monitored through foliar relative water content (TRA) and pre-dawn (ypd) and midday water potential (ymd). Gas exchanges, chlorophyll a fluorescence and photosynthetic capacity (Amax) were measured. Pigments and soluble protein were quantified and antioxidant system enzymes activity was determined. Plants under water deficit were rehydrated and the same measurements and sampling were done again after 48 h (R1) and 72 h (R2). Albeit water availability has diminished from 86% to 47% of soil capacity, there weren’t any remarkable changes on S plants. R1 plants which haven’t shown any alterations neither in Amax nor in stomatal conductance (gH2O), exhibited a significant decrease on net photosynthesis (A), reaching negative values, along with a raise in non-photochemical quenching (NPQ). Presumably this raise should be related with an increase in the xanthophyll cycle deepoxidation state. The significantly increase in Superoxide dismutase (SOD) specific activity in R1 and R2 plants might be related with the activation of Mehler-peroxidase reaction which provokes lumen DpH raising. This can provide photosynthesis protection through VAZ cycle and may justify the NPQ increase.
- Fotossíntese e acumulação de ácido abcísico em Lupinus albus sujeito a stress hídricoPublication . Correia, Maria João; David, Maria Manuela; Barrote, IsabelThe objective of the present work was to assess whether the depression of photosynthetic activity in droughted Lupinus albus L. plants was more closely associated with carbohydrate build-up or ABA accumulation. With that purpose we have measured the concomitant drought-induced changes in photosynthetic capacity and the concentrations of Rubisco, chlorophylls, non-structural carbohydrates and ABA in young and old leaves of white lupin plants. Although Rubisco and chlorophylls contents did not decrease with water stress, the photosynthetic capacity was decreased by soil drying, the decline in photosynthesis being more tighly related with ABA accumulation than with sugar content. In contrast to droughted plants, ABA feeding to intact plants resulted in a significant decrease in Rubisco and chlorophyll content. However, no increase in shoot ABA content was detected in ABA-fed plants. Therefore this work is not conclusive as to the correlation between photosynthetic capacity and ABA accumulation in droughted white lupin plants being causal or not.
- 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.
- Interaction between water and nutrient deficiencies in helianthus annuusPublication . Fonseca, Filomena; David, Maria Manuela; Barrote, Isabel; Ferreira, Gisela; Correia, Maria João; Martins-Loução, M. A.; Hips, S. H.The physiological response of plants to water deficits are known to vary according to the conditions of application of drought stress and the rate of development of leaf water deficits. At the whole plant level the effect of the water shess is usually perceived as a decrease in photosynthesis and growth, and is associated with alterations in C and N metabolism (McDonald and Davies, 1996). The decrease in water potential affects transpiration and hence xylem transport of nitrate or reduced N into growing regions. The response of the photo-synthetic apparatus either to water stress or rehydration seems to be dependent "on leaf age (O'Neill, 1983; Wolfe et al., 1988). Degradation of both thylakoid and stromal N-containing compounds can occur in response to water stress, recovery from which may pequire more than a week (Chaves, 1991).