Dehydration and rehydration differently affect photosynthesis and volatile monoterpenes in bryophytes with contrasting ecological traits.

Bryophytes desiccate rapidly when relative humidity decreases. The capacity to withstand dehydration depends on several ecological and physiological factors. Volatile organic compounds (VOCs) may have a role in enhancing tolerance to desiccating bryophytes. However, the functions of VOCs in bryophytes have received little attention so far. We aimed to investigate the impact of a dehydration-rehydration treatment on primary carbon metabolism and volatile terpenes (VTs) in three bryophytes with contrasting ecological traits: Vessicularia dubyana, Porella platyphylla and Pleurochaete squarrosa. First, we confirmed the desiccation sensitivity gradient of the species. Under fully hydrated conditions, the photosynthetic rate (A) was inversely associated with stress tolerance, with a lower rate in more tolerant species. The partial recovery of A in P. platyphylla and P. squarrosa after rehydration confirmed the desiccation tolerance of these two species. On the other hand, A did not recover after rehydration in V. dubyana. Regarding VT, each species exhibited a distinct VT profile under optimum hydration, with the highest VT pool found in the more desiccation-sensitive species (V. dubyana). However, the observed species-specific VT pattern could be associated with the ecological habitat of each species. P. squarrosa, a moss of dry habitats, may synthesize mainly non-volatile secondary metabolites as stress-defensive compounds. On the other hand, V. dubyana, commonly found submerged, may need to invest photosynthetically assimilated carbon to synthesize a higher amount of VTs to cope with transient water stress occurrence. Further research on the functions of VTs in bryophytes is needed to deepen our understanding of their ecological significance.

Acquisition of Desiccation Tolerance Unveiled: Polar Lipid Profiles of Streptophyte Algae Offer Insights.

Terrestrialization by photosynthetic eukaryotes took place in the two branches of green microalgae: Chlorophyta and Charophyta. Within the latter, the paraphyletic streptophytic algae divide into two clades. These are named Klebsormidiophyceae-Chlorokybophyceae-Mesostigmatophyceae (KCM), which is the oldest, and Zygnematophyceae-Coleochaetophyceae-Charophyceae (ZCC), which contains the closest relatives of vascular plants. Terrestrialization required the emergence of adaptations in response to new challenges, such as irradiance, temperature oscillations and water deprivation. In this study, we evaluated lipid composition in species representative of distinct phylogenetic clusters within Charophyta and Chlorophyta. We aim to study whether the inherent thylakoid lipid composition, as well as its adaptability in response to desiccation, were fundamental factors for the evolutionary history of terrestrial plants. The results showed that the lipid composition was similar to that found in flowering land plants, differing only in betaine lipids. Likewise, the largest constitutive pool of oligogalactolipids (OGL) was found only in the fully desiccation-tolerant species Klebsormidium nitens. After desiccation, the content of polar lipids decreased in all species. Conversely, the content of OGL increased, particularly trigalactosyldiacylglycerol and tetragalactosyldiacylglycerol in the ZCC clade. The analysis of the molecular species composition of the newly formed OGL may suggest a different biosynthetic route for the KCM and ZCC clades. We speculate that the appearance of a new OGL synthesis pathway, which eventually arose during the streptophyte evolutionary process, endowed algae with a much more dynamic regulation of thylakoid composition in response to stress, which ultimately contributed to the colonization of terrestrial habitats.

Freezing induces an increase in leaf spectral transmittance of forest understorey and alpine forbs.

Evergreen plants growing at high latitudes or high elevations may experience freezing events in their photosynthetic tissues. Freezing events can have physical and physiological effects on the leaves which alter leaf optical properties affecting remote and proximal sensing parameters. We froze leaves of six alpine plant species (Soldanella alpina, Ranunculus kuepferi, Luzula nutans, Gentiana acaulis, Geum montanum, and Centaurea uniflora) and three evergreen forest understorey species (Hepatica nobilis, Fragaria vesca and Oxalis acetosella), and assessed their spectral transmittance and optically measured pigments, as well as photochemical efficiency of photosystem II (PSII) as an indicator of freezing damage. Upon freezing, leaves of all the species transmitted more photosynthetically active radiation (PAR) and some species had increased ultraviolet-A (UV-A) transmittance. These differences were less pronounced in alpine than in understorey species, which may be related to higher chlorophyll degradation, visible as reduced leaf chlorophyll content upon freezing in the latter species. Among these understorey forbs, the thin leaves of O. acetosella displayed the largest reduction in chlorophyll (-79%). This study provides insights into how freezing changes the leaf optical properties of wild plants which could be used to set a baseline for upscaling optical reflectance data from remote sensing. Changes in leaf transmittance may also serve to indicate photosynthetic sufficiency and physiological tolerance of freezing events, but experimental research is required to establish this functional association.

Protective Strategies of Haberlea rhodopensis for Acquisition of Freezing Tolerance: Interaction between Dehydration and Low Temperature.

Resurrection plants are able to deal with complete dehydration of their leaves and then recover normal metabolic activity after rehydration. Only a few resurrection species are exposed to freezing temperatures in their natural environments, making them interesting models to study the key metabolic adjustments of freezing tolerances. Here, we investigate the effect of cold and freezing temperatures on physiological and biochemical changes in the leaves of Haberlea rhodopensis under natural and controlled environmental conditions. Our data shows that leaf water content affects its thermodynamical properties during vitrification under low temperatures. The changes in membrane lipid composition, accumulation of sugars, and synthesis of stress-induced proteins were significantly activated during the adaptation of H. rhodopensis to both cold and freezing temperatures. In particular, the freezing tolerance of H. rhodopensis relies on a sucrose/hexoses ratio in favor of hexoses during cold acclimation, while there is a shift in favor of sucrose upon exposure to freezing temperatures, especially evident when leaf desiccation is relevant. This pattern was paralleled by an elevated ratio of unsaturated/saturated fatty acids and significant quantitative and compositional changes in stress-induced proteins, namely dehydrins and early light-induced proteins (ELIPs). Taken together, our data indicate that common responses of H. rhodopensis plants to low temperature and desiccation involve the accumulation of sugars and upregulation of dehydrins/ELIP protein expression. Further studies on the molecular mechanisms underlying freezing tolerance (genes and genetic regulatory mechanisms) may help breeders to improve the resistance of crop plants.

Shedding light on the dark side of xanthophyll cycles.

Xanthophyll cycles are broadly important in photoprotection, and the reversible de-epoxidation of xanthophylls typically occurs in excess light conditions. However, as presented in this review, compiling evidence in a wide range of photosynthetic eukaryotes shows that xanthophyll de-epoxidation also occurs under diverse abiotic stress conditions in darkness. Light-driven photochemistry usually leads to the pH changes that activate de-epoxidases (e.g. violaxanthin de-epoxidase), but in darkness alternative electron transport pathways and luminal domains enriched in monogalactosyl diacyl glycerol (which enhance de-epoxidase activity) likely enable de-epoxidation. Another 'dark side' to sustaining xanthophyll de-epoxidation is inactivation and/or degradation of epoxidases (e.g. zeaxanthin epoxidase). There are obvious benefits of such activity regarding stress tolerance, and indeed this phenomenon has only been reported in stressful conditions. However, more research is required to unravel the mechanisms and understand the physiological roles of dark-induced formation of zeaxanthin. Notably, the de-epoxidation of violaxanthin to antheraxanthin and zeaxanthin in darkness is still a frequently ignored process, perhaps because it questions a previous paradigm. With that in mind, this review seeks to shed some light on the dark side of xanthophyll de-epoxidation, and point out areas for future work.

Alpine forbs rely on different photoprotective strategies during spring snowmelt.

Snowmelt in alpine ecosystems brings ample water, and together with above-freezing temperatures, initiates plant growth. In this scenario, rapid activation of photosynthesis is essential for a successful life-history strategy. But, strong solar radiation in late spring enhances the risk of photodamage, particularly before photosynthesis is fully functional. We compared the photoprotective strategy of five alpine forbs: one geophyte not particularly specialised in subnival life (Crocus albiflorus) and four wintergreens differing in their degree of adaptation to subnival life, from least to most specialised: Gentiana acaulis, Geum montanum, Homogyne alpina and Soldanella alpina. We used distance to the edge of snow patches as a proxy to study time-dependent changes after melting. We postulated that the photoprotective response of snowbed specialists would be stronger than of more-generalist alpine meadow species. Fv /Fm was relatively low across wintergreens and even lower in the geophyte C. albiflorus. This species also had the largest xanthophyll-cycle pool and lowest tocopherol and flavonoid glycoside contents. After snow melting, all the species progressively activated ETR, but particularly the intermediate snowbed species G. acaulis and G. montanum. The photoprotective responses after snowmelt were idiosyncratic: G. montanum rapidly accumulated xanthophyll-cycle pigments, tocopherol and flavonoid glycosides; while S. alpina showed the largest increase in plastochromanol-8 and chlorophyll contents and the greatest changes in optical properties. Climate warming scenarios might shift the snowmelt date and consequently alter the effectiveness of photoprotection mechanisms, potentially changing the fitness outcome of the different strategies adopted by alpine forbs.

Born to revive: molecular and physiological mechanisms of double tolerance in a paleotropical and resurrection plant.

Resurrection plants recover physiological functions after complete desiccation. Almost all of them are native to tropical warm environments. However, the Gesneriaceae include four genera, remnant of the past palaeotropical flora, which inhabit temperate mountains. One of these species is additionally freezing-tolerant: Ramonda myconi. We hypothesise that this species has been able to persist in a colder climate thanks to some resurrection-linked traits. To disentangle the physiological mechanisms underpinning multistress tolerance to desiccation and freezing, we conducted an exhaustive seasonal assessment of photosynthesis (gas exchange, limitations to partitioning, photochemistry and galactolipids) and primary metabolism (through metabolomics) in two natural populations at different elevations. R. myconi displayed low rates of photosynthesis, largely due to mesophyll limitation. However, plants were photosynthetically active throughout the year, excluding a reversible desiccation period. Common responses to desiccation and low temperature involved chloroplast protection: enhanced thermal energy dissipation, higher carotenoid to Chl ratio and de-epoxidation of the xanthophyll cycle. As specific responses, antioxidants and secondary metabolic routes rose upon desiccation, while putrescine, proline and a variety of sugars rose in winter. The data suggest conserved mechanisms to cope with photo-oxidation during desiccation and cold events, while additional metabolic mechanisms may have evolved as specific adaptations to cold during recent glaciations.

A field portable method for the semi-quantitative estimation of dehydration tolerance of photosynthetic tissues across distantly related land plants.

Desiccation tolerant (DT) plants withstand complete cellular dehydration, reaching relative water contents (RWC) below 30% in their photosynthetic tissues. Desiccation sensitive (DS) plants exhibit different degrees of dehydration tolerance (DHT), never surviving water loss >70%. To date, no procedure for the quantitative evaluation of DHT extent exists that is able to discriminate DS species with differing degrees of DHT from truly DT plants. We developed a simple, feasible and portable protocol to differentiate between DT and different degrees of DHT in the photosynthetic tissues of seed plants and between fast desiccation (< 24 h) tolerant (FDT) and sensitive (FDS) bryophytes. The protocol is based on (1) controlled desiccation inside Falcon tubes equilibrated at three different relative humidities that, consequently, induce three different speeds and extents of dehydration and (2) an evaluation of the average percentage of maximal photochemical efficiency of PSII (Fv /fm) recovery after rehydration. Applying the method to 10 bryophytes and 28 tracheophytes from various locations, we found that (1) imbibition of absorbent material with concentrated salt-solutions inside the tubes provides stable relative humidity and avoids direct contact with samples; (2) for 50 ml capacity tubes, the optimal plant amount is 50-200 mg fresh weight; (3) the method is useful in remote locations due to minimal instrumental requirements; and (4) a threshold of 30% recovery of the initial Fv /fm upon reaching RWC ≤ 30% correctly categorises DT species, with three exceptions: two poikilochlorophyllous species and one gymnosperm. The protocol provides a semi-quantitative expression of DHT that facilitates comparisons of species with different morpho-physiological traits and/or ecological attributes.

Can Parietin Transfer Energy Radiatively to Photosynthetic Pigments?

The main role of lichen anthraquinones is in protection against biotic and abiotic stresses, such as UV radiation. These compounds are frequently deposited as crystals outside the fungal hyphae and most of them emit visible fluorescence when excited by UV. We wondered whether the conversion of UV into visible fluorescence might be photosynthetically used by the photobiont, thereby converting UV into useful energy. To address this question, thalli of Xanthoria parietina were used as a model system. In this species the anthraquinone parietin accumulates in the outer upper cortex, conferring the species its characteristic yellow-orange colouration. In ethanol, parietin absorbed strongly in the blue and UV-B and emitted fluorescence in the range 480⁻540 nm, which partially matches with the absorption spectra of photosynthetic pigments. In intact thalli, it was determined by confocal microscopy that fluorescence emission spectra shifted 90 nm towards longer wavelengths. Then, to study energy transfer from parietin, we compared the response to UV of untreated and parietin-free thalli (removed with acetone). A chlorophyll fluorescence kinetic assessment provided evidence of UV-induced electron transport, though independently of the presence of parietin. Thus, a role for anthraquinones in energy harvesting is not supported for X. parietina under presented experimental conditions.

Endogenous circadian rhythms in pigment composition induce changes in photochemical efficiency in plant canopies.

There is increasing evidence that the circadian clock is a significant driver of photosynthesis that becomes apparent when environmental cues are experimentally held constant. We studied whether the composition of photosynthetic pigments is under circadian regulation, and whether pigment oscillations lead to rhythmic changes in photochemical efficiency. To address these questions, we maintained canopies of bean and cotton, after an entrainment phase, under constant (light or darkness) conditions for 30-48 h. Photosynthesis and quantum yield peaked at subjective noon, and non-photochemical quenching peaked at night. These oscillations were not associated with parallel changes in carbohydrate content or xanthophyll cycle activity. We observed robust oscillations of Chl a/b during constant light in both species, and also under constant darkness in bean, peaking when it would have been night during the entrainment (subjective nights). These oscillations could be attributed to the synthesis and/or degradation of trimeric light-harvesting complex II (reflected by the rhythmic changes in Chl a/b), with the antenna size minimal at night and maximal around subjective noon. Considering together the oscillations of pigments and photochemistry, the observed pattern of changes is counterintuitive if we assume that the plant strategy is to avoid photodamage, but consistent with a strategy where non-stressed plants maximize photosynthesis.

Diversity of winter photoinhibitory responses: a case study in co-occurring lichens, mosses, herbs and woody plants from subalpine environments.

Winter evergreens living in mountainous areas have to withstand a harsh combination of high light levels and low temperatures in wintertime. In response, evergreens can activate a photoprotective process that consists of the downregulation of photosynthetic efficiency, referred to as winter photoinhibition (WPI). WPI has been studied mainly in woody evergreens and crops even when, in many instances, other functional groups such as lichens or bryophytes dominate in alpine and boreal habitats. Thus, we aimed to (1) assess the occurrence of WPI within overwintering evergreens comprising woody species, herbs, mosses and lichens, (2) compare the recovery kinetics among those groups and (3) clarify the role of thylakoid proteins and pigments in both processes: WPI and recovery. With this aim, WPI was analyzed in 50 species in the field and recovery kineticcs were studied in one model species from each functional group. Results showed that high levels of WPI are much more frequent among woody plants than in any other group, but are also present in some herbs, lichens and mosses. Winter conditions almost always led to the de-epoxidation of the xanthophyll cycle. Nevertheless, changes in the de-epoxidation level were not associated with the activation/deactivation of WPI in the field and did not match changes in photochemical efficiency during recovery treatments. Seasonal changes in thylakoid proteins [mainly D1 (photosystem II core complex protein) and PsbS (essential protein for thermal dissipation)] were dependent on the functional group. The results highlight the diversity of physiological solutions and suggest a physical-mechanical reason for the more conservative strategy of woody species compared with other groups.

Emissions of carotenoid cleavage products upon heat shock and mechanical wounding from a foliose lichen.

Carotenoids constitute a major target of chloroplastic photooxidative reactions, leading to the formation of several oxidized derivatives and cleavage products, some of which are volatile (VCCPs). Among them, ß-cyclocitral (ß-CC), at least, is a retrograde signaling molecule that modulates the activity of many key physiological processes. In the present work, we aimed to study whether ß-CC and other VCCPs are released into the atmosphere from photosynthetic tissues. To overcome stomatal limitations, the foliose chlorolichen Lobaria pulmonaria was used as the model system, and the emissions of biogenic volatiles, induced by heat and wounding stresses, were monitored by proton-transfer reaction time-of-flight mass-spectrometry (PTR-TOF-MS) and gas-chromatography (GC-MS). Prior to stress treatments, VCCPs were emitted constitutively, accounting for 1.3 % of the total volatile release, with ß-CC being the most abundant VCCP. Heat and wounding stresses induced a burst of volatile release, including VCCPs, and a loss of carotenoids. Under heat stress, the production of ß-CC correlated positively with temperature. However the enhancement of production of VCCPs was the lowest among all the groups of volatiles analyzed. Given that the rates of carotenoid loss were three orders of magnitude higher than the release rates of VCCPs and that these compounds only represent a minor fraction in the blend of volatiles, it seems unlikely that VCCPs might represent a global stress signal capable of diffusing through the atmosphere to different neighboring individuals.

Internal and external factors affecting photosynthetic pigment composition in plants: a meta-analytical approach.

Photosynthetic pigment composition has been a major study target in plant ecophysiology during the last three decades. Although more than 2000 papers have been published, a comprehensive evaluation of the responses of photosynthetic pigment composition to environmental conditions is not yet available. After an extensive survey, we compiled data from 525 papers including 809 species (subkingdom Viridiplantae) in which pigment composition was described. A meta-analysis was then conducted to assess the ranges of photosynthetic pigment content. Calculated frequency distributions of pigments were compared with those expected from the theoretical pigment composition. Responses to environmental factors were also analysed. The results revealed that lutein and xanthophyll cycle pigments (VAZ) were highly responsive to the environment, emphasizing the high phenotypic plasticity of VAZ, whereas neoxanthin was very stable. The present meta-analysis supports the existence of relatively narrow limits for pigment ratios and also supports the presence of a pool of free 'unbound' VAZ. Results from this study provide highly reliable ranges of photosynthetic pigment contents as a framework for future research on plant pigments.

Activation of photoprotective winter photoinhibition in plants from different environments: a literature compilation and meta-analysis.

Overwintering plants face a pronounced imbalance between light capture and use of that excitation for photosynthesis. In response, plants upregulate thermal dissipation, with concomitant reductions in photochemical efficiency, in a process characterized by a slow recovery upon warming. These sustained depressions of photochemical efficiency are termed winter photoinhibition (WPI) here. WPI has been extensively studied in conifers and in few overwintering crops, but other plant species have received less attention. Furthermore, the literature shows some controversies about the association of WPI with xanthophylls and the environmental conditions that control xanthophylls conversion. To overview current knowledge and identify knowledge gaps on WPI mechanisms, we performed a comprehensive meta-analysis of literature published over the period 1991-2011. All publications containing measurements of Fv/Fm for a cold period and a corresponding warm control were included in our final database of 190 studies on 162 species. WPI was estimated as the relative decrease in Fv/Fm. High WPI was always accompanied by a high (A + Z)/(V + A + Z). Activation of lasting WPI was directly related to air temperature, with a threshold of around 0°C. Tropical plants presented earlier (at a temperature of >0°C) and higher WPI than non-tropical plants. We conclude that (1) activation of a xanthophyll-dependent mechanism of WPI is a requisite for maintaining photosynthetic structures at sub-zero temperatures, while (2) absence (or low levels) of WPI is not necessarily related to low (A + Z)/(V + A + Z); and (3) the air temperature that triggers lasting WPI, and the maximum level of WPI, do not depend on plant growth habit or bioclimatic origin of species.

Side-effects of domestication: cultivated legume seeds contain similar tocopherols and fatty acids but less carotenoids than their wild counterparts.

BACKGROUND: Lipophilic antioxidants play dual key roles in edible seeds (i) as preservatives of cell integrity and seed viability by preventing the oxidation of fats, and (ii) as essential nutrients for human and animal life stock. It has been well documented that plant domestication and post-domestication evolution frequently resulted in increased seed size and palatability, and reduced seed dormancy. Nevertheless, and surprisingly, it is poorly understood how agricultural selection and cultivation affected the physiological fitness and the nutritional quality of seeds. Fabaceae have the greatest number of crop species of all plant families, and most of them are cultivated for their highly nutritious edible seeds. Here, we evaluate whether evolution of plants under cultivation has altered the integrated system formed by membranes (fatty acids) and lipophilic antioxidants (carotenoids and tocopherols), in the ten most economically important grain legumes and their closest wild relatives, i.e.: Arachis (peanut), Cicer (chickpea), Glycine (soybean), Lathyrus(vetch), Lens (lentil), Lupinus (lupin), Phaseolus (bean), Pisum (pea), Vicia (faba bean) and Vigna (cowpea). RESULTS: Unexpectedly, we found that following domestication, the contents of carotenoids, including lutein and zeaxanthin, decreased in all ten species (total carotenoid content decreased 48% in average). Furthermore, the composition of carotenoids changed, whereby some carotenoids were lost in most of the crops. An undirected change in the contents of tocopherols and fatty acids was found, with contents increasing in some species and decreasing in others, independently of the changes in carotenoids. In some species, polyunsaturated fatty acids (linolenic acid especially), α-tocopherol and γ-tocopherol decreased following domestication. CONCLUSIONS: The changes in carotenoids, tocopherols and fatty acids are likely side-effects of the selection for other desired traits such as the loss of seed dormancy and dispersal mechanisms, and selection for seed storability and taste. This work may serve as baseline to broaden our knowledge on the integrated changes on crop fitness and nutritional quality following domestication.

Evidence for the absence of enzymatic reactions in the glassy state. A case study of xanthophyll cycle pigments in the desiccation-tolerant moss Syntrichia ruralis.

Desiccation-tolerant plants are able to withstand dehydration and resume normal metabolic functions upon rehydration. These plants can be dehydrated until their cytoplasm enters a 'glassy state' in which molecular mobility is severely reduced. In desiccation-tolerant seeds, longevity can be enhanced by drying and lowering storage temperature. In these conditions, they still deteriorate slowly, but it is not known if deteriorative processes include enzyme activity. The storage stability of photosynthetic organisms is less studied, and no reports are available on the glassy state in photosynthetic tissues. Here, the desiccation-tolerant moss Syntrichia ruralis was dehydrated at either 75% or <5% relative humidity, resulting in slow (SD) or rapid desiccation (RD), respectively, and different residual water content of the desiccated tissues. The molecular mobility within dry mosses was assessed through dynamic mechanical thermal analysis, showing that at room temperature only rapidly desiccated samples entered the glassy state, whereas slowly desiccated samples were in a 'rubbery' state. Violaxanthin cycle activity, accumulation of plastoglobules, and reorganization of thylakoids were observed upon SD, but not upon RD. Violaxanthin cycle activity critically depends on the activity of violaxanthin de-epoxidase (VDE). Hence, it is proposed that enzymatic activity occurred in the rubbery state (after SD), and that in the glassy state (after RD) no VDE activity was possible. Furthermore, evidence is provided that zeaxanthin has some role in recovery apparently independent of its role in non-photochemical quenching of chlorophyll fluorescence.

Gesneriads, a Source of Resurrection and Double-Tolerant Species: Proposal of New Desiccation- and Freezing-Tolerant Plants and Their Physiological Adaptations.

Gesneriaceae is a pantropical family of plants that, thanks to their lithophytic and epiphytic growth forms, have developed different strategies for overcoming water scarcity. Desiccation tolerance or "resurrection" ability is one of them: a rare phenomenon among angiosperms that involves surviving with very little relative water content in their tissues until water is again available. Physiological responses of desiccation tolerance are also activated during freezing temperatures, a stress that many of the resurrection gesneriads suffer due to their mountainous habitat. Therefore, research on desiccation- and freezing-tolerant gesneriads is a great opportunity for crop improvement, and some of them have become reference resurrection angiosperms (Dorcoceras hygrometrica, Haberlea rhodopensis and Ramonda myconi). However, their difficult indoor cultivation and outdoor accessibility are major obstacles for their study. Therefore, this review aims to identify phylogenetic, geoclimatic, habitat, and morphological features in order to propose new tentative resurrection gesneriads as a way of making them more reachable to the scientific community. Additionally, shared and species-specific physiological responses to desiccation and freezing stress have been gathered as a stress response metabolic basis of the family.

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.

Physiology of the seasonal relationship between the photochemical reflectance index and photosynthetic light use efficiency.

The photochemical reflectance index (PRI) is regarded as a promising proxy to track the dynamics of photosynthetic light use efficiency (LUE) via remote sensing. The implementation of this approach requires the relationship between PRI and LUE to scale not only in space but also in time. The short-term relationship between PRI and LUE is well known and is based on the regulative process of non-photochemical quenching (NPQ), but at the seasonal timescale the mechanisms behind the relationship remain unclear. We examined to what extent sustained forms of NPQ, photoinhibition of reaction centres, seasonal changes in leaf pigment concentrations, or adjustments in the capacity of alternative energy sinks affect the seasonal relationship between PRI and LUE during the year in needles of boreal Scots pine. PRI and NPQ were highly correlated during most of the year but decoupled in early spring when the foliage was deeply downregulated. This phenomenon was attributed to differences in the physiological mechanisms controlling the seasonal dynamics of PRI and NPQ. Seasonal adjustments in the pool size of the xanthophyll cycle pigments, on a chlorophyll basis, controlled the dynamics of PRI, whereas the xanthophyll de-epoxidation status and other xanthophyll-independent mechanisms controlled the dynamics of NPQ at the seasonal timescale. We conclude that the PRI leads to an underestimation of NPQ, and consequently overestimation of LUE, under conditions of severe stress in overwintering Scots pine, and most likely also in species experiencing severe drought. This severe stress-induced decoupling may challenge the implementation of the PRI approach.

Assessing Plant Pigment Regulation in Circadian Experiments.

Circadian rhythms affect many aspects of a plant's metabolism including, but not limited to, photosynthesis. Here, we provide a complete protocol for determining changes in the composition of photosynthetic pigments (chlorophyll and carotenoids), and we also consider its implementation within circadian experiments. We describe how to design a circadian experiment with the goal of assessing changes in pigment composition. We then perform two consecutive approaches to track changes in pigment composition: indirect noninvasive estimation of pigment composition (by reflectance or fluorescence) followed by direct pigment analysis (by chromatography or spectrophotometry). Finally, we present several considerations regarding data analyses.