Raquel Esteban

Thermal energy dissipation and xanthophyll cycles beyond the Arabidopsis model

Thermal dissipation of excitation energy is a fundamental photoprotection mechanism in plants. Thermal energy dissipation is frequently estimated using the quenching of the chlorophyll fluorescence signal, termed non-photochemical quenching. Over the last two decades, great progress has been made in the understanding of the mechanism of thermal energy dissipation through the use of a few model plants, mainly Arabidopsis. Nonetheless, an emerging number of studies suggest that this model represents only one strategy among several different solutions for the environmental adjustment of thermal energy dissipation that have evolved among photosynthetic organisms in the course of evolution. In this review, a detailed analysis of three examples highlights the need to use models other than Arabidopsis: first, overwintering evergreens that develop a sustained form of thermal energy dissipation; second, desiccation tolerant plants that induce rapid thermal energy dissipation; and third, understorey plants in which a complementary lutein epoxide cycle modulates thermal energy dissipation. The three examples have in common a shift from a photosynthetically efficient state to a dissipative conformation, a strategy widely distributed among stress-tolerant evergreen perennials. Likewise, they show a distinct operation of the xanthophyll cycle. Expanding the list of model species beyond Arabidopsis will enhance our knowledge of these mechanisms and increase the synergy of the current studies now dispersed over a wide number of species.

Photoprotective implications of leaf variegation in E. dens-canis L. and P. officinalis L

Variegated leaves occur rarely in nature, but there are some species, primarily in the forest understory, that possess this characteristic. We recently studied two variegated plants: Erytronium dens-canis L., which is characterised by a pattern of red patches and Pulmonaria officinalis L., with light green spots. These non-green areas could attenuate light reaching mesophyll cells with respect to green sections. The aim of the study was to verify whether such red and light green parts are more photoprotected than green ones and if this trait could be of adaptive value. Red patches in E. dens-canis were due to a single layer of red cells in the upper parenchyma, which accumulated anthocyanins. Light green spots in P. officinalis were caused by the presence of loosely arranged cells instead of a well-established layer of packed cells in the palisade parenchyma. Chlorophyll fluorescence imaging was performed under light treatment, showing a greater decrease of photochemical efficiency in red and light green patches than in green sections. Differences in the extent of photochemical efficiency among patches were not attributable to different activation of the xanthophyll cycle. These observations failed to confirm our initial hypothesis, but they questioned the physiological reason for this higher sensitivity in red and light green patches of photosynthetic tissues. Chlorophyll fluorescence imaging was therefore performed in the field. The same pattern of photochemical efficiency was maintained only in E. dens-canis. The current results demonstrate that in both species the benefits of variegation, if any, are different from enhanced photosynthetic performance.

Distribution and evolutionary trends of photoprotective isoprenoids (xanthophylls and tocopherols) within the plant kingdom.

The earliest land photosynthesis would have increased the risk of photo-oxidations and the demand of anti-oxidative protection. In this work, we aimed to determine the evolutionary trends in photoprotection across a wide representation of the plant kingdom and to verify whether the non-ubiquitous lutein-epoxide (Lx) cycle is a polyphyletic or an ancient character. Carotenoids and alpha-tocopherol (alpha-toc) were analysed by HPLC in 266 species. Phylogenetic analyses of the presence of photoprotective compounds and zeaxanthin-epoxidase (ZE) sequences were performed. Violaxanthin-cycle pigments (VAZ) and alpha-toc were taxonomically ubiquitous. Ancient groups showed higher contents of VAZ than vascular plants, while alpha-toc showed the opposite pattern. Lutein-epoxide was present in 45% of the species. It showed a remarkable variation across groups but with a clear increasing trend from algae to basal angiosperms. Lutein-epoxide was also related to woody trait and leaf longevity. No correlation between the presence of Lx and recurrent mutations in ZE sequences, including the duplications, was found. Thus, there is an evolutionary trend to increase the content of alpha-toc and to decrease the total amount of VAZ pigments. Absence of Lx in algae discards an ancestral origin. Present results are also inconsistent with a polyphyletic origin of Lx in angiosperms.

Physical factors driving intertidal macroalgae distribution: physiological stress of a dominant fucoid at its southern limit

Climate change is driving species range shifts worldwide. However, physiological responses related to distributional changes are not fully understood. Oceanographers have reported an increase in ocean temperature in the northwest Iberian Peninsula that is potentially related to the decline in some cold-temperate intertidal macroalgae in the Cantabrian Sea, namely Fucus serratus. Low tide stress could also play a role in this decline. We performed one mensurative (in situ) and two manipulative (in culture) experiments designed to evaluate the interactive effects of some physical factors. The first experiment analysed field response to low tide stress in marginal (mid-Cantabrian Sea and northern Portugal) versus central (Galicia) populations of F. serratus. Then a second experiment was performed that utilized either harsh or mild summer conditions of atmospheric temperature, irradiance, humidity, and wind velocity to compare the responses of individuals from one marginal and one central population to low tide stress. Finally, the combined effect of sea temperature and the other factors was evaluated to detect interactive effects. Changes in frond growth, maximal photosynthetic quantum yield (Fv/Fm), temperature, and desiccation were found. Three additive factors (solar irradiation, ocean and air temperatures) were found to drive F. serratus distribution, except under mildly humid conditions that ameliorated atmospheric thermal stress (two additive factors). Mid-Cantabrian Sea temperatures have recently increased, reaching the inhibitory levels suggested in this study of F. serratus. We also expect an additive secondary contribution of low tide stress to this species decline. On the northern Portugal coast, ocean warming plus low tide stress has not reached this species’ inhibition threshold. No significant differential responses attributed to the population of origin were found. Mechanistic approaches that are designed to analyse the interactive effects of physical stressors may improve the levels of confidence in predicted range shifts of species.

Effectiveness of arbuscular mycorrhizal fungi (AMF) for inducing the accumulation of major carotenoids, chlorophylls and tocopherol in green and red leaf lettuces

Previous studies demonstrated that arbuscular mycorrhizal fungi (AMF) can induce the accumulation of carotenoids, phenolics, anthocyanins and some mineral nutrients in leaves of lettuce (Lactuca sativa L.) thus enhancing its nutritional quality. Our objectives were to know which carotenoids were the most accumulated in leaves of mycorrhizal lettuces and to assess the effect of AMF on tocopherols’ levels in leaves of lettuce plants. AMF always enhanced growth and, in most cases, increased the levels of all major carotenoids, chlorophylls and tocopherols in green and red leaf lettuces. Since these molecules are also important nutraceuticals, mycorrhization emerges as reliable technique to enhance the nutritional value of edible vegetables. These results are compared with other methods developed to improve nutritional quality.

Photoprotective responses of Mediterranean and Atlantic trees to the extreme heat-wave of summer 2003 in Southwestern Europe

Summer 2003 was extremely hot in Europe. High light in combination with heat and drought exacerbates the generation of photo-oxidative stress. Under these conditions photoprotective responses can be critical for plant survival. Photoprotection was analysed in 2003 in several Mediterranean and Atlantic woody species. These data were compared with previous summers (1998, 1999 and 2001) to evaluate the potential acclimation for each species. A pattern of changes consisting on a decrease in chlorophyll, ascorbate and Fv/Fm and an increase in tocopherol, xanthophyll cycle pigments (VAZ) and de-epoxidation index was regularly observed. Acclimation potential was measured by the use of the plasticity index for each parameter. Mediterranean species were more plastic than Atlantic ones. The latter were unable to increase antioxidant pools to the same extent or to down-regulate the efficiency of light energy conversion. These results indicate that most Mediterranean species are able to perform an efficient acclimation to heat stress, whilst Atlantic species will be more affected by climate warming.

Dark induction of the photoprotective xanthophyll cycle in response to dehydration.

Some plants tolerate tissue dehydration. Dehydration conditions suppress photosynthesis, exacerbating photooxidative stress. In this study, fern samples were collected from the field, desiccated in darkness, and subsequently re-watered. During dark dehydration, zeaxanthin (Z) was formed and maximal photochemical efficiency of PS II was strongly reduced. Rehydration in the dark reversed these effects. Violaxanthin de-epoxidase was responsible for the dark formation of Z as illustrated by its complete inhibition by DTT. Nonetheless, its activity was not affected by nigericin, indicating that Z formation in the dark could be a process independent of the transmembrane pH-gradient into the thylakoids. Synthesis de novo of Z was rejected after blocking carotenogenesis with norfluorazon. Dark formation of Z was also observed in dehydrating leaves of desiccation-intolerant plants, which seems to indicate that this is a phenomenon scattered among different taxa within the plant kingdom. Plants may trigger this mechanism during dehydration, for chlorophyll protection during desiccation, and for faster acclimation when rehydrating conditions return. Violaxanthin de-epoxidation to form Z is typically a light-dependent process, but the formation induced solely by dehydration might represent an anticipatory mechanism for preventing early morning photodamage in desiccation-tolerant plants such as the fern Ceterach officinarum.

Carotenoid composition in Rhodophyta: insights into xanthophyll regulation in Corallina elongata

Macroalgae possess several photoprotection mechanisms, including xanthophyll cycles. Among these cycles, the VAZ (violaxanthin–anteraxanthin–zeaxanthin) cycle controls the interconversion of epoxidated xanthophyll (V) to de-epoxidated forms anteraxanthin and zeaxanthin. This conversion modulates the amount of excess energy that it is dissipated as heat. Presence of functional xanthophyll cycles in Rhodophyta is controversial. In this work we investigated the presence of xanthophyll cycles in red algae. Carotenoid composition of 13 red macroalgae collected in northern Spain was studied by HPLC. This screening showed that carotenoid composition is a conservative trade in rhodophytes, with very few exceptions to the general pattern formed by β-carotene and one to three xanthophylls: anteraxanthin, zeaxanthin or lutein. Corallina elongata and Jania rubens were the only algae containing anteraxanthin as the main xanthophyll. The first species was selected to study whether any truncated xanthophyll cycle between anteraxanthin and zeaxanthin could operate under photoinhibitory conditions. Upon illumination, xanthophyll composition remained stable, without any short-term light-induced de-epoxidation of anteraxanthin to zeaxanthin. However, an inverse relation between both xanthophylls was found. In addition, a seasonal trend of changes in anteraxanthin and zeaxanthin was observed when pigment composition was studied in field samples over the course of a year. We concluded that these variations were more likely due to differential rates of synthesis and degradation of xanthophylls than to the operation of a xanthophyll cycle. The great amount of anteraxanthin in C. elongata may play an structural stability role on light-harvesting complexes.

High irradiance induces photoprotective mechanisms and a positive effect on NH4+ stress in Pisum sativum L.

Photosynthesis provides plant metabolism with reduced carbon (C) but is also the main source of oxidative stress in plants. Likewise, high doses of NH(4)(+) as sole N source have been reported to be toxic for most plants, resulting in reduced plant growth and restricting C availability. The combination of high photosynthetic photon flux densities (PPFD) and NH(4)(+) nutrition may provide higher C availability but could also have a detrimental effect on the plants, therefore the objective of this study is to evaluate whether NH(4)(+) induces photo-oxidative stress that is exacerbated under high light conditions. Pea plants (Pisum sativum cv. sugar-snap) were grown hydroponically with NH(4)(+) (0.5, 2.5, 5 and 10 mM) under high (750 micromol photons m(-2)s(-1)) or low PPFD conditions (350 micromol photons m(-2)s(-1)). High PPFD contributes to a higher tolerance to ammonium by pea plants, as it originated higher biomass content due to higher photosynthetic rates. However, a deficit of N (0.5 and 2.5 mM NH(4)(+)) under high PPFD conditions caused an antioxidant response, as indicated by increased photoprotective pigment and chloroplastic superoxide dismutase contents. Plants grown with higher doses of N and high PPFD showed less need for photoprotection. An increase in the specific leaf weight (SLW) ratio was observed associated not only with high PPFDs but also with the highest NH(4)(+) dose. Overall, these results demonstrate that, despite the activation of some photoprotective responses at high PPFD, there were no photoinhibitory symptoms and a positive effect on NH(4)(+) toxicity, thus suggesting that the harmful effects of NH(4)(+) are not directly related to the generation of photo-oxidative stress.

Short‐ and long‐term modulation of the lutein epoxide and violaxanthin cycles in two species of the Lauraceae: sweet bay laurel (Laurus nobilis L.) and avocado (Persea americana Mill.)

Short- and long-term responses of the violaxanthin (V) and lutein epoxide (Lx) cycles were studied in two species of Lauraceae: sweet bay laurel (Laurus nobilis L.) and avocado (Persea americana L.). The Lx content exceeded the V content in shade leaves of both species. Both Lx and V were de-epoxidised on illumination, but only V was fully restored by epoxidation in low light. Violaxanthin was preferentially de-epoxidised in low light in L. nobilis. This suggests that Lx accumulates with leaf ageing, partly because its conversion to lutein is limited in shade. After exposure to strong light, shade leaves of avocado readjusted the total pools of alpha- and beta-xanthophyll cycles by de novo synthesis of antheraxanthin, zeaxanthin and lutein. This occurred in parallel with a sustained depression of F(v)/F(m). In Persea indica, a closely related but low Lx species, F(v)/F(m) recovered faster after a similar light treatment, suggesting the involvement of the Lx cycle in sustained energy dissipation. Furthermore, the seasonal correlation between non-reversible Lx and V photoconversions and pre-dawn F(v)/F(m) in sun leaves of sweet bay supported the conclusion that the Lx cycle is involved in a slowly reversible downregulation of photosynthesis analogous to the V cycle.