Modified atmosphere packaging and dark/light refrigerated storage in green leafy vegetables have impact on nutritional value

The consumption of zeaxanthin (Z) through a vegetable-rich diet is recommended to reduce the progression of age-related macular degeneration. Due to Z’s intrinsic dynamic character that results from its participation in the photoprotective xanthophyll cycle involving the carotenoids violaxanthin, antheraxanthin and zeaxanthin (VAZ), post-harvest handling practices and storage usually retain low amounts of this bioactive compound (compared to the rest of phytochemicals that are, in general, more stable). Thus, the aim of this work was to investigate in important consumed leafy vegetables the effects of different storage conditions on carotenoids (mainly Z) including i) packaging under three modified atmospheres (MAs), ii) light refrigerated supermarket storage and iii) dark refrigerated domestic storage. The results showed that an MA with low O2 and high CO2 enhanced the Z content under light. Moreover, both light and dark refrigerated storage showed dynamic and circadian pigment changes that enhanced the total VAZ pool. These results can contribute to generating practical recommendations for industries, supermarkets, and consumers when high Z content is a nutritional target.RE received a Juan de la Cierva-incorporación grant IJCI-2014-21452. JB is a holder of a PhD fellowship from the Public University of Navarre. This research was supported by research BFU 2010-15021 and CTM2014- 53902-C2-2-P from the Ministry of Education and Science of Spain and the ERDF (FEDER) and research project UPV/EHU IT-1018-16. Technical and human support by Eroski supermarkets and “Fundación Tecnova” is gratefully acknowledged

Activation of violaxanthin cycle in darkness is a common response to different abiotic stresses: a case study in Pelvetia canaliculata

<p>Abstract</p> <p>Background</p> <p>In the violaxanthin (V) cycle, V is de-epoxidized to zeaxanthin (Z) when strong light or light combined with other stressors lead to an overexcitation of photosystems. However, plants can also suffer stress in darkness and recent reports have shown that dehydration triggers V-de-epoxidation in the absence of light. In this study, we used the highly stress-tolerant brown alga <it>Pelvetia canaliculata </it>as a model organism, due to its lack of lutein and its non-photochemical quenching independent of the transthylakoidal-ΔpH, to study the triggering of the V-cycle in darkness induced by abiotic stressors.</p> <p>Results</p> <p>We have shown that besides desiccation, other factors such as immersion, anoxia and high temperature also induced V-de-epoxidation in darkness. This process was reversible once the treatments had ceased (with the exception of heat, which caused lethal damage). Irrespective of the stressor applied, the resulting de-epoxidised xanthophylls correlated with a decrease in Fv/Fm, suggesting a common function in the down-regulation of photosynthetical efficiency. The implication of the redox-state of the plastoquinone-pool and of the differential activity of V-cycle enzymes on V-de-epoxidation in darkness was also examined. Current results suggest that both violaxanthin de-epoxidase (VDE) and zeaxanthin-epoxidase (ZE) have a basal constitutive activity even in darkness, being ZE inhibited under stress. This inhibition leads to Z accumulation.</p> <p>Conclusion</p> <p>This study demonstrates that V-cycle activity is triggered by several abiotic stressors even when they occur in an absolute absence of light, leading to a decrease in Fv/Fm. This finding provides new insights into an understanding of the regulation mechanism of the V-cycle and of its ecophysiological roles.</p

Xanthophyll cycles in the juniper haircap moss (Polytrichum juniperinum) and Antarctic hair grass (Deschampsia antarctica) on Livingston Island (South Shetland Islands, Maritime Antarctica)

The summer climate in Maritime Antarctica is characterised by high humidity and cloudiness with slightly above zero temperatures. Under such conditions, photosynthetic activity is temperature-limited and plant communities are formed by a few species. These conditions could prevent the operation of the photoprotective xanthophyll (VAZ) cycle as low irradiance reduces the excess of energy and low temperatures limit enzyme activity. The VAZ cycle regulates the dissipation of the excess of absorbed light as heat, which is the main mechanism of photoprotection in plants. To test whether this mechanism operates dynamically in Antarctic plant communities, we characterised pigment dynamics under natural field conditions in two representative species: the moss Polytrichum juniperinum and the grass Deschampsia antarctica. Pigment analyses revealed that the total VAZ pool was in the upper range of the values reported for most plant species, suggesting that they are exposed to a high degree of environmental stress. Despite cloudiness, there was a strong conversion of violaxanthin (V) to zeaxanthin (Z) during daytime. Conversely, the dark-induced enzymatic epoxidation back to V was not limited by nocturnal temperatures. In contrast with plants from other cold ecosystems, we did not find any evidence of overnight retention of Z or sustained reductions in photochemical efficiency. These results are of interest for modelling, remote sensing and upscaling of the responses of Antarctic vegetation to environmental challenges.The Spanish Ministry of Science, Innovation and Universities (MICIU/FEDER, EU) and the Basque Government funded this research through the projects CTM2014-53902-C2-2-P, PGC2018-093824-B-C44 and UPV/EHU IT-1018-16. Open Access funding provided thanks to the CRUE-CSIC agreement with Springer Nature

More than just lipid balls: quantitative analysis of plastoglobule attributes and their stress‑related responses

[EN] Main conclusion Plastoglobules are ubiquitous under non-stress conditions and their morphology, closely related to their composition, changes differently depending on the specific stress that the plant undergoes. Plastoglobules are lipoprotein structures attached to thylakoid membranes, which participate in chloroplast metabolism and stress responses. Their structure contains a coating lipid monolayer and a hydrophobic core that differ in composition. Their function in chloroplasts has been studied focussing on their composition. However, we currently lack a comprehensive study that quantitatively evaluates the occurrence and morphology of plastoglobules. Following a literature search strategy, we quantified the main morphological attributes of plastoglobules from photosynthetic chloroplasts of more than 1000 TEM images published over the last 53 years, covering more than 100 taxa and 15 stress types. The analysis shows that plastoglobules under non-stress conditions are spherical, with an average diameter of 100-200 nm and cover less than 3% of the chloroplast cross-section area. This percentage rises under almost every type of stress, particularly in senescence. Interestingly, an apparent trade-off between increasing either the number or the diameter of plastoglobules governs this response. Our results show that plastoglobules are ubiquitous in chloroplasts of higher plants under non-stress conditions. Besides, provided the specific molecular composition of the core and coat of plastoglobules, we conclude that specific stress-related variation in plastoglobules attributes may allow inferring precise responses of the chloroplast metabolism.This work was supported by the project PGC2018-093824-B-C44 from the Ministerio de Ciencia, Innovación y Universidades (MCIU, Spain) and the European Regional Development Fund (ERDF) and grant UPV/EHU IT-1018-16 from the Basque Government (Spain). MIA received a pre-doctoral grant from the Basque Government. We thank J. Miranda-Apodaca for helpful assistance with PCA analysis. Open Access funding provided thanks to the CRUE-CSIC agreement with Springer Nature

Rapid colour changes in Euglena sanguinea (Euglenophyceae) caused by internal lipid globule migration

The accumulation of red pigments under chronic stress is a response observed in most groups of oxygenic photoautotrophs. It is thought that the red pigments in the cell shield the chlorophyll located underneath from the light. Among these red pigments, the accumulation of carotenoids is one of the most frequent cases. However, the synthesis or degradation of carotenoids is a slow process and this response is usually only observed when the stress is maintained over a period of time. In the Euglenophyte Euglena sanguinea, this is due to the accumulation of a large amount of free and esterified astaxanthin (representing 80% of the carotenoid pool). While reddening is a slow and sometimes irreversible process in other phototrophs, reducing the efficiency of light harvesting by chlorophyll, in E. sanguinea it is highly dynamic, capable of shifting from red to green (and vice-versa) in 10-20 min. This change is not due to de novo carotenogenesis, but to the relocation of cytoplasmic lipid globules where astaxanthin accumulates. Thus, red globules migrate from the centre of the cell to peripheral locations when photoprotection is demanded. This protective system seems to be so efficient that other classical mechanisms are not operative in this species. For example, despite the presence and operation of the diadino-diatoxanthin cycle, nonphotochemical quenching (NPQ) is almost undetectable. Since E. sanguinea forms extensive floating colonies, reddening can be observed at much greater scale than at a cellular level, the mechanism described here being one of the fastest and most dramatic colour changes attributable to photosynthetic organisms at cell and landscape level. In sum, these data indicate an extremely dynamic and efficient photoprotective mechanism based on organelle migration more than on carotenoid biosynthesis that prevents excess light absorption by chlorophylls reducing the need for other protective processes related to energy dissipation.This work was supported by the Basque Government [UPV/EHU-GV IT-1018-16] [UPV/EHU PPG17/67 – GV IT-1040-16], and by the Spanish Ministry of Economy and Competitiveness (MINECO) and the European Research and Development Foundation (FEDER) through (i) [CTM2014-53902-C2-2-P] national grant and (ii) a “Juan de la Cierva-Incorporación” postdoctoral grant [IJCI-2014-22489] to BFM

Unravelling metabolic mechanisms behind chloroplast desiccation tolerance: chlorophyllous fern spore as a new promising unicellular model

Se sustituye el preprint por el postprint el 23-11-2020Fern spores are unicellular structures produced by the sporophyte generation that give rise to the haploid gametophyte. When released from the sporangium, spores are desiccation tolerant (DT) in the royal fern (Osmunda regalis) and contain fully developed chloroplasts. As a consequence this type of spores are called chlorophyllous spores (CS). Upon transfer to germination conditions, CS initiate a process of imbibition that suppress DT in 72h, before the germination starts. In parallel to such change in DT, thylakoids undergo a profound remodelling in composition and function. Firstly, sustained quenching of chlorophyll fluorescence is relaxed, giving rise to photochemically active CS, while lipid composition shifts from that of a resting structure to a metabolically active cell. Basically trigalactolipids decreased in favour of monogalactolipids, with a parallel desaturation of fatty acids. Storage lipids such as triacylglycerol were quickly depleted. These results highlight the importance of the structure of thylakoids lipid as a key to protect membrane integrity during desiccation, together with the saturation of fatty acids and the constitutive chlorophyll quenching to prevent oxidative damage. The CS used here, in which the same cell shifts from DT to sensitive strategy in 72h, reveal its their potential as unicellular models for future studies on DT.This work was funded by (i) the Basque Government (research project UPV/EHU IT-1018-16; and Predoctoral Fellowship to MLP); (ii) the Spanish Ministry of Economy and Competitiveness (MINECO) and the ERDF (FEDER) (research project CTM2014-53902-C2-2-P, and Juan de la Cierva-Incorporation fellowship IJCI-2014-22489 to BFM)

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.Marie Curie IEF grant (328370 MELISSA) from the Euro- pean FP7-PEOPLE and a ‘Juan de la Cierva Incorporación’ grant (IJCI-2014-22489) from the Spanish Ministry of Econ- omy and Competitiveness (MINECO) and the European Regional Development Fund ERDF (FEDER). The work was also supported by the project BFU 2010 – 15021 and CTM2014-53902-C2-2-P from the MINECO and the ERDF (FEDER), and by the Basque Government (UPV/EHU-GV IT-624–13 and UPV/EHU-GV IT-1018-16

Photosynthetic responses of trees in high-elevation forests: comparing evergreen species along an elevation gradient in the Central Andes

Plant growth at extremely high elevations is constrained by high daily thermal amplitude, strong solar radiation and water scarcity. These conditions are particularly harsh in the tropics, where the highest elevation treelines occur. In this environment, the maintenance of a positive carbon balance involves protecting the photosynthetic apparatus and taking advantage of any climatically favourable periods. To characterize photoprotective mechanisms at such high elevations, and particularly to address the question of whether these mechanisms are the same as those previously described in woody plants along extratropical treelines, we have studied photosynthetic responses in Polylepis tarapacana Philippi in the central Andes (18 degrees S) along an elevational gradient from 4300 to 4900 m. For comparative purposes, this gradient has been complemented with a lower elevation site (3700 m) where another Polylepis species (P. rugulosa Bitter) occurs. During the daily cycle, two periods of photosynthetic activity were observed: one during the morning when, despite low temperatures, assimilation was high; and the second starting at noon when the stomata closed because of a rise in the vapour pressure deficit and thermal dissipation is prevalent over photosynthesis. From dawn to noon there was a decrease in the content of antenna pigments (chlorophyll b and neoxanthin), together with an increase in the content of xanthophyll cycle carotenoids. These results could be caused by a reduction in the antenna size along with an increase in photoprotection. Additionally, photoprotection was enhanced by a partial overnight retention of de-epoxized xanthophylls. The unique combination of all of these mechanisms made possible the efficient use of the favourable conditions during the morning while still providing enough protection for the rest of the day. This strategy differs completely from that of extratropical mountain trees, which uncouple light-harvesting and energy-use during long periods of unfavourable, winter conditions.This research was carried out with the aid of grants from the Chilean Research Council (FONDECYT 1120965 and FONDAP 15110009) awarded to D.A.C., and BFU 2010-15021 from the Spanish Ministry of Economy and Competitiveness (MINECO) and the European Regional Development Fund ERDF(FEDER) and the Basque Government (UPV/EHU-GV IT-299-07) awarded to J.I.G.-P

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.Peer reviewe

Unraveling the Photoprotective Response of Lichenized and Free-Living Green Algae (Trebouxiophyceae, Chlorophyta) to Photochilling Stress

Lichens and free-living terrestrial algae are widespread across many habitats and develop successfully in ecosystems where a cold winter limits survival. With the goal of comparing photoprotective responses in free-living and lichenized algae, the physiological responses to chilling and photochilling conditions were studied in three lichens and their isolated algal photobionts together as well as in a fourth free-living algal species. We specifically addressed the following questions: (i) Are there general patterns of acclimation in green algae under chilling and photochilling stresses? (ii) Do free-living algae exhibit a similar pattern of responses as their lichenized counterparts? (iii) Are these responses influenced by the selection pressure of environmental conditions or by the phylogenetic position of each species? To answer these questions, photosynthetic fluorescence measurements as well as pigment and low molecular weight carbohydrate pool analyses were performed under controlled laboratory conditions. In general, photochemical efficiency in all free-living algae decreased with increasing duration of the stress, while the majority of lichens maintained an unchanged photochemical activity. Nevertheless, these patterns cannot be generalized because the alga Trebouxia arboricola and the lichen Ramalina pollinaria (associated with Trebouxia photobionts) both showed a similar decrease in photochemical efficiency. In contrast, in the couple Elliptochloris bilobata-Baeomyces rufus, only the algal partner exhibited a broad physiological performance under stress. This study also highlights the importance of the xanthophyll cycle in response to the studied lichens and algae to photochilling stress, while the accumulation of sugars was not related to cold acclimation, except in the alga E. bilobata. The differences in response patterns detected among species can be mainly explained by their geographic origin, although the phylogenetic position should also be considered, especially in some species.FM received a fellowship for PhD students (2012-2015) from the Government of the Basque Country as well as a mobility grant which allowed the collaboration between University of Basque Country (UPV/EHU) and University of Rostock. LG thanks the Deutsche Forschungsgemeinschaft for financial support (DFG GU 1278/1-1). The work was also supported by the project BFU 2010-15021 and CTM2014-53902-C2-2-P from the MINECO and the European Regional Development Fund ERDF (FEDER), and by the Basque Government (UPV/EHU-GV IT-624-13 and UPV/EHU-GV IT-1018-16)