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- Effectiveness and consistency of a suite of descriptors to assess the ecological status of seagrass meadows (Posidonia oceanica L. Delile)Publication . Rotini, Alice; Belmonte, A.; Barrote, Isabel; Micheli, C.; Peirano, A.; Santos, Rui; Silva, JoãoThe increasing rate of human-induced environmental changes on coastal marine ecosystems has created a demand for effective descriptors, in particular for those suitable for monitoring the status of seagrass meadows. Growing evidence has supported the useful application of biochemical and genetic descriptors such as secondary metabolite synthesis, photosynthetic activity and genetic diversity. In the present study, we have investigated the effectiveness of different descriptors (traditional, biochemical and genetic) in monitoring seagrass meadow conservation status. The Posidonia oceanica meadow of Monterosso al Mare (Ligurian sea, NW Mediterranean) was subjected to the measurement of bed density, leaf biometry, total phenols, soluble protein and photosynthetic pigment content as well as to RAPD marker analysis. This suite of descriptors provided evidence of their effectiveness and convenient application as markers of the conservation status of P. oceanica and/or other seagrasses. Biochemical/genetic descriptors and those obtained by traditional methods depicted a well conserved meadow with seasonal variability and, particularly in summer, indicated a healthier condition in a portion of the bed (station C), which was in agreement with the physical and sedimentological features of the station. Our results support the usefulness of introducing biochemical and genetic approaches to seagrass monitoring programs since they are effective indicators of plant physiological stress and environmental disturbance.
- Physiological stress indicators in seagrassesPublication . Barrote, Isabel; Santos, Rui; Silva, JoãoWe investigated the combined effects of several environmental stressors in the biochemical defence mechanisms of the intertidal seagrass Zostera noltii and the subtidal seagrasses Zostera marina and Cymodocea nodosa in Ria Formosa coastal lagoon (southern Portugal). During one year Z. noltii was sampled for antioxidant enzymes activity, pigments, soluble protein and malondialdehyde (MDA) in both neap and spring tides. Intertidal seagrasses are periodically exposed to potentially stressful conditions such as desiccation, osmotic stress, high light and very high or freezing temperatures, depending on the season, latitude and tidal amplitude. In Ria Formosa Z. marina and C. nodosa appear frequently in the same meadow and compete for space, light and nutrients. These species are also exposed to stressful environmental conditions such as light or temperature stress and their survival depends on their relative ability to cope with all these stressors. In order to study the effects of low light in Z. marina and C. nodosa, four different levels of shading were imposed on field plots of both species during three weeks. P-I relationships were determined and samples were collected to determine photosynthetic pigments, non-structural carbohydrates, soluble protein, phenols and malondialdehyde concentration. Reactive oxygen species (ROS) are formed as a normal part of the plants metabolism. An increment on ROS formation is a common response to stress and can induce several types of damage, namely lipid peroxidation. MDA is a product of the peroxidation of membrane lipids and thus is commonly used as an indicator of oxidative stress, as is the concentration of phenolic compounds. Carotenoids and antioxidant enzymes are part of the plants’ antioxidative system. Higher content of carotenoids, besides amplifying the photosynthetic useful wavelengths, indicate higher photoprotection. Increased activities of ROS scavenging enzymes are correlated with stress tolerance. The relationships among experimental and environmental data will be discussed.
- Effects of in situ shading on the photophysiology of Zostera marina and Cymodocea nodosaPublication . Silva, João; Barrote, Isabel; Albano, Sílvia; Costa, Monya; Neves, Pedro; Graça, Gonçalo Nuno Santos Neto da; Sharon, Yoni; Beer, S.; Santos, RuiThe effects of light reduction were investigated in natural stands of the seagrasses Zostera marina and Cymodocea nodosa in Ria Formosa coastal lagoon, southern Portugal. Four shading plots and a control were set in each of two neighbouring meadows (2-3 m depth), each dominated by one species. The experiment lasted for 3 weeks, at the end of which the response of plant photosynthesis to light was determined via oxygen electrode measurements. Tissue samples were also analysed for photosynthetic pigment, soluble protein, soluble sugar and malondialdehyde contents. All plants presented a shade-adapted profile, mostly revealed by their biochemical composition. In both species the chlorophyll a/chlorophyll b ratio decreased sharply whereas the total chlorophyll/total carotenoids and the total chlorophyll/soluble protein ratios increased. Soluble protein content was reduced more noticeably in Z. marina. Soluble sugars dropped almost 40% in Z. marina leaves and roots, a more pronounced effect than the one observed in C. nodosa. Overall, Z. marina revealed to be more sensitive than C. nodosa to reductions in incident irradiance, suggesting that it will be more sensitive to human-induced disturbances that result in an increase of water turbidity.
- The effects of epiphytes on light harvesting and antioxidant responses in the seagrass posidonia oceanicaPublication . Costa, Monya; Silva, João; Olivé, Irene; Barrote, Isabel; Alexandre, Ana; Albano, Sílvia; Santos, RuiPosidonia oceanica (L.) Delile is a subtidal seagrass whose leaves are commonly colonized by epiphytes. Epiphytes pose physical barriers to light penetration within the leaves, with possible significant impacts on photosynthesis. Furthermore, epiphytes can indirectly be responsible for leaf chlorosis, necrosis and senescence which are known to be related with the increase of oxygen reactive species (ROS) levels, potentially leading to oxidative stress. The aim of this work was to investigate in situ (i) the effect of epiphytes on the composition and balance of light harvesting pigments in leaves of the naturally growing seagrass P. oceanica, and (ii) evaluate differences in antioxidant responses. Epiphytized and non-epiphytized plants were analyzed to establish potential photosynthetic pigment roleshift between light harvesting and photoprotection functions. The experiments were carried out in Cabo de Gata Natural Park, southern Spain, where epiphytized and non-epiphytized plants can be found at identical depths and light exposure. The results showed that both O2 evolution rate along the day and chlorophyll a/b ratio were higher in non-epiphytized plants, indicating a negative effect of epiphytes on photosynthesis and light harvesting. Although under high irradiance (at solar noon) the xanthophyll cycle was activated in both epiphytized and non-epiphytized leaves, the de-epoxidation-ratio (AZ/VAZ) was lower in epiphytized leaves, due to light attenuation by epiphytes. The antioxidant capacity (TEAC and ORAC essays) and the activity of the antioxidant enzymes ascorbate peroxidase and dehydroascorbate were higher in epiphytized plants, showing that epiphytes can also be a potential source of oxidative stress to P. oceanica. Our results show that despite the light attenuation effect, leaf colonization by epiphytes can also be potentially stressful and reduces plant productivity.
- Physiological responses of Zostera marina and Cymodocea nodosa to light-limitation stressPublication . Silva, João; Barrote, Isabel; Costa, Monya; Albano, Sílvia; Santos, RuiThe effects of light-limitation stress were investigated in natural stands of the seagrasses Zostera marina and Cymodocea nodosa in Ria Formosa coastal lagoon, southern Portugal. Three levels of light attenuation were imposed for 3 weeks in two adjacent meadows (2–3 m depth), each dominated by one species. The response of photosynthesis to light was determined with oxygen electrodes. Chlorophylls and carotenoids were determined by high-pressure liquid chromatography (HPLC). Soluble protein, carbohydrates, malondialdehyde and phenol contents were also analysed. Both species showed evident signs of photoacclimation. Their maximum photosynthetic rates were significantly reduced with shading. Ratios between specific light harvesting carotenoids and the epoxidation state of xanthophyll cycle carotenoids revealed significantly higher light harvesting efficiency of C. nodosa, a competitive advantage in a low light environment. The contents of both soluble sugars and starch were considerably lower in Z. marina plants, particularly in the rhizomes, decreasing even further with shading. The different carbohydrate energy storage strategies found between the two species clearly favour C. nodosa's resilience to light deprivation, a condition enhanced by its intrinsic arrangement of the pigment pool. On the other hand, Z. marina revealed a lower tolerance to light reduction, mostly due to a less plastic arrangement of the pigment pool and lower carbohydrate storage. Our findings indicate that Z. marina is close to a light-mediated ecophysiological threshold in Ria Formosa.
- Seasonal photosynthesis, respiration, and calcification of a temperate Maërl bed in southern PortugalPublication . SORDO, LAURA; Santos, Rui; Barrote, Isabel; Freitas, Cátia; Silva, JoãoRhodolith (maerl) beds are biodiversity hotspots with a worldwide distribution. Maerl is the general term for free-living non-geniculate rhodoliths or coralline red algae. In southern Portugal, maerl beds are mainly composed of Phymatolithon lusitanicum, recently identified as a new species and commonly misidentified as Phymatolithon calcareum. Photosynthesis, respiration, and growth rates of the algae were measured seasonally, as well as the photosynthetic pigment composition. To characterize the seasonal and interannual patterns of key abiotic conditions in the largest described maerl bed of the Portuguese coast, temperature, irradiance, and dissolved oxygen were continuously monitored over a 2-year period. At the bed depth (22 m), temperature ranged between 14 degrees C in winter and 24 degrees C in summer, irradiance varied from 5 to 75 mu.mol m(-2) s(-1) , and dissolved oxygen from 5.8 to 7.25 mg O-2 L-1. We found a strong linear relationship (r(2) = 0.95) between gross primary production (GPP) and relative electron transport rates (rETRs). Both methods led to similar results and an average molar ratio of 0.24. Photosynthesis and respiration increased in summer and decreased in autumn and winter. In the summer of 2013, the growth rates were twofold higher (1.34 mu.mol CaCO3 g(-1) day(-1)) than in the other seasons. In winter and spring, to compensate for light deprivation and low temperature, algae increased their chlorophyll a and carotenoid concentrations while also decreasing their phycobilin concentration, in this case probably due to nutrient limitation. To isolate the role of temperature on the algae's metabolism, the photosynthetic and respiration rates of individual thalli were measured at eight different temperatures in the laboratory (from 12 degrees C to 26 degrees C). Phymatolithon lusitanicum photosynthesis increased twofold after a threshold of 18 degrees C (from 2.2 at 18 degrees C to 3.87 mu mol O-2 m(-2) s(-1) at 20 degrees C), whereas respiration increased fourfold with temperature after a threshold of 22 degrees C (from -0.38 at 18 degrees C to -1.81 (mu mol O-2 m(-2) s(-1) at 24 degrees C). The significant increases on respiration, photosynthetic rates, and maximum growth with temperature reveal that the metabolic rates of P. lusitanicum are highly sensitive to ocean warming.
- Compared photophysiology of native seagrasses with an invasive macroalga in Sydney Harbour, AustraliaPublication . Silva, João; Runcie, J. W.; Barrote, Isabel; Costa, Monya; Santos, RuiThe comparative photophysiology of the seagrasses Zostera capricorni and Halophila ovalis and the invasive macroalgae Caulerpa taxifolia was investigated in a shallow mixed meadow at Chowder Bay (Sydney, Australia), where the three species coexist. This study was developed under the auspices of the COST Action “Seagrasses: from genes to ecosystems”, targeting one of its objectives, the development of innovative devices for the continuous measurement of seagrass photosynthesis. Automated multi-channel chlorophyll fluorometers were deployed for 24-hour periods to examine and compare the changes in the photosynthetic efficiency and energy quenching mechanisms of the 3 species. Tissue samples were collected at predawn and noon, frozen and analysed by HPLC for detailed pigment analysis. The invasive C. taxifolia showed generally higher photosynthetic efficiency than both Z. capricorni and H. ovalis. Both seagrass species showed down-regulation of photosynthesis at noon, evidenced by the low quantum yield and the significant reduction of the antennae pigments between pre-dawn and noon. In contrast, C. taxifolia showed no reduction in antennae pigments or total photosynthetic pigments along the day. While both seagrasses showed 3- to 7-fold increases in the epoxidation state of xanthophyll cycle pigments between pre-dawn and noon, in C. taxifolia there was no significant change. Our results show that C. taxifolia is better adapted to high light conditions than Z. capricorni and H. ovalis, which means that, from the photophysiological point of view, this invasive macroalgae constitutes a serious competitor for seagrasses in shallow areas with high irradiance.