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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.

Abscisic acid (ABA) is an important carotenoid-derived phytohormone that plays essential roles in plant response to biotic and abiotic stresses as well as in various physiological and developmental processes. In Arabidopsis, ABA biosynthesis starts with the epoxidation of zeaxanthin by the ABA DEFICIENT 1 (ABA1) enzyme, leading to epoxycarotenoids, e.g., violaxanthin. The oxidative cleavage of 9-cis-epoxycarotenoids, a key regulatory step catalyzed by 9-CIS-EPOXYCAROTENOID DIOXYGENASE, forms xanthoxin that is converted in further reactions mediated by ABA DEFICIENT 2 (ABA2), ABA DEFICIENT 3 (ABA3), and ABSCISIC ALDEHYDE OXIDASE 3 (AAO3) into ABA. By combining genetic and biochemical approaches, we unravel here an ABA1-independent ABA biosynthetic pathway starting upstream of zeaxanthin. We identified the carotenoid cleavage products, i.e., apocarotenoids, β-apo-11-carotenal, 9-cis-β-apo-11-carotenal, 3-OH-β-apo-11-carotenal, and 9-cis-3-OH-β-apo-11-carotenal as intermediates of this ABA1-independent ABA biosynthetic pathway. Using labeled compounds, we showed that β-apo-11-carotenal, 9-cis-β-apo-11-carotenal, and 3-OH-β-apo-11-carotenal are successively converted into 9-cis-3-OH-β-apo-11-carotenal, xanthoxin, and finally into ABA in both Arabidopsis and rice. When applied to Arabidopsis, these β-apo-11-carotenoids exert ABA biological functions, such as maintaining seed dormancy and inducing the expression of ABA-responsive genes. Indeed, the transcriptomic analysis revealed a high overlap of differentially expressed genes regulated by β-apo-11-carotenoids and ABA, but also suggested that these compounds exert ABA-independent regulatory activities. Taken together, our study identifies a biological function for the common plant metabolites β-apo-11-carotenoids, extends our knowledge about ABA biosynthesis and provides new insights into plant apocarotenoid metabolic networks.

The aim of this study was to evaluate the effects of melatonin on photoinhibition under chilling stress in cucumber seedlings and to inquire into any mechanisms of mitigation. Under chilling stress, the net photosynthetic rate declined dramatically but the decline was significantly mitigated by irrigation with a melatonin solution. Possible mechanisms for this mitigation are that melatonin accelerates xanthophyll de-epoxidation by upregulating the transcription of the violaxanthin de-epoxidase gene (CsVDE) and down-regulating that of the zeaxanthin cyclase gene (CsZE) during chilling. There was also a rise in non-photochemical quenching (NPQ) if seedlings were pretreated with melatonin before chilling. The efficient operation of the xanthophyll cycle helped consume excessive excitation energy in photosystem (PS) Ⅱ and so protected the photosynthetic system. Melatonin also modulated the partitioning of absorbed excitation energy in PS Ⅱ as evidenced by alleviation of the decrease in quantum yield of photochemical energy conversion in PS Ⅱ under chilling stress, by alleviation of the rise in quantum yield of non-regulated, non-photochemical energy loss in PS II and by increasing the regulated non-photochemical energy loss in PS II. This study presents a new understanding of the mechanisms through which melatonin mitigates photoinhibition by modulating the partitioning of absorption energy in PS Ⅱ based on the xanthophyll cycle.

The interest in bioactive compounds from microalgae is increasing since they have medicinal and nutritional areas. The present work aims to evaluate the potential pharmaceutical interest of extracts from three eustigmatophyte strains from the Coimbra Collection of Algae (ACOI): Chlorobotrys gloeothece, Chlorobotrys regularis and Characiopsis aquilonaris. Antioxidant and antiproliferative activities were determined as well as chlorophyll a, carotenoid and phenolic total contents. In addition, major pigments and sterols were identified and quantified. The three strains were grown until the stationary phase and then the biomass was extracted. Antioxidant activity was measured by TEAC, DPPH and FRAP assays and antiproliferative effect was assessed by the MTT method on MCF-7, PC-3 and NHDF cells. The pigment and phenolic total contents were determined by spectrophotometry. Of these strains, C. aquilonaris showed the highest antioxidant activity measured by TEAC and FRAP assays (23.98 ± 0.01 μmol TE eq g−1 DW and 42.57 ± 0.04 μmol TE eq g−1 DW, respectively), a selective effect in reduting MCF-7 cells proliferation and a larger amount of chlorophyll a, carotenoids and phenolic content (18.40 ± 0.00 μg chlorophyll a mg−1 DW, 2.27 ± 0.00 mg carotenoids g−1 DW and 6.23 ± 0.01 mg GAE g−1 DW, respectively). A positive correlation between chlorophyll a and TEAC assay was observed, as well as between carotenoids and TEAC and FRAP assays, suggesting these compounds as important contributors to significant antioxidant activity. Violaxanthin, cholesterol and stigmasterol were present in larger amount in C. aquilonaris while C. regularis showed a higher amount of β-carotene. These results suggest that these three ACOI eustigmatophytes are promising for applications in the improvement of human health, particularly in cancer prevention and treatment.

This study aimed to optimize the key parameters of extraction methods and to increase the recovery yields of intact xanthophylls (violaxanthin, zeaxanthin, astaxanthin) from microalgae (Chlorella luteoviridis). An effective, simple, and fast extraction protocol is described. It consists of a grinding pretreatment followed by a microwave-assisted extraction, using ethanol 90% as an environmentally preferable extraction solvent. Xanthopylls were quantified using high performance liquid chromatography. Irradiation time of 6 s only resulted in the extraction of violaxanthin (4.479 ± 0.009 mg/g), astaxanthin (4.154 ± 0.013 mg/g), and zeaxanthin (4.776 ± 0.120 mg/g). The described protocol seems to be the fastest extraction method of xantophylls compared to the literature and could be an advantage for industrial scale, while saving time and energy.

The toxic potential of Chattonella is associated with a high production of reactive oxygen species (ROS). Chattonella species can tolerate high irradiance levels but seems not to be efficient in the induction of nonphotochemical chl a fluorescence (NPQ) under light stress conditions. Therefore, we postulated that high ROS production of this microalgal group is related to the lack of effective photoprotection mechanisms. We compared the NPQ induction, xanthophyll cycle interconversion (XC), and the production of the ion superoxide (O2 - ) in Chattonella marina var. antiqua, Chattonella sp., and C. marina acclimated to 43 (LL) and 300 µmol photons · m2 · s-1 (HL). We also evaluated the photosynthetic characteristics of the three strains. Photosynthesis saturated at relative high irradiances (above 500 µmol photons · m2 · s-1 ) in LL and HL Chattonella strains. For the first time, we documented the conversion of diadinoxanthin into diatoxanthin in microalgae that have violaxanthin as the major XC carotenoid. The slow NPQ induction indicated that qE (fast component of NPQ) was not present, and this process was related to the interconversion of XC pigments. However, the quenching efficiency (QE) of deepoxidated xanthophylls was low in the three Chattonella strains. The strain with the lowest QE produced the highest amount of a O2 - . Therefore, ROS production in Chattonella seems to be related to a low expression of XC-related thermal PSII dissipation.

We examined light quality-dependent regulation of photoprotection and antioxidant properties in rice seedlings. Seedlings grown under red or green light-emitting diodes (LEDs) showed declines in quantum yield of electron transport through PSII, maximum relative electron transport rates, and photochemical quenching compared to the white and blue LED, but no difference in maximum photochemical efficiency of PSII was found. Blue LED treatment resulted in greater increases in not only nonphotochemical quenching, but also zeaxanthin formation, expressions of β-carotene hydroxylase and violaxanthin de-epoxidase, and anthocyanin compared to other LED treatments. By contrast, green and red LEDs increased activities of superoxide dismutase and ascorbate peroxidase as well as expression levels of catalase (CAT)a and CATb to white and blue LEDs. Green and red LEDs also greatly upregulated expressions of phytochrome (PHY) A, PHYB, PHYC, and cryptochrome 2 compared to white and blue LEDs, whereas blue LED upregulated CRY1. Our results demonstrate that specific wavelengths of different LEDs differentially influence photosynthetic performance, photoprotection, and antioxidant properties, possibly through regulating expressions of phytochrome and cryptochrome genes in a light-quality-dependent manner.

Petal colour is considered as an important trait and major visual signal for insect‐pollinated plants and landscaping. In previous study, we isolated and identified two genes (BnPC1 and BnPC2) that control the orange petal colour (OPC) in rapeseed. To identify the molecular markers linked to the orange petal colour, we constructed a B. napus linkage map with four newly developed simple sequence repeat (SSR) markers and nine insertion–deletion (InDel) markers using BC₅II, a near isogenic line population segregated at the BnPC2 locus. The BnPC2 gene was flanked by Indel‐3 and Indel‐20 and was found to represent a 27‐kb (3.721–3.748 Mb) region on A09 of B. napus. High‐pressure liquid chromatography analysis revealed that, compared to yellow petals, orange petals exhibit significantly increased zeaxanthin, β‐carotene and lutein contents and a relatively decreased violaxanthin content. Expression profiling confirmed that BnaA09.ZEP (zeaxanthin epoxidase) is an important candidate gene for BnPC2 and is predominantly expressed in floral tissues. Overall, our results provide a basis for understanding the molecular mechanisms underlying the orange petal colour trait in B. napus.

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.

Pigments are a fundamental part of the microalgal cell, with chlorophylls at the center of photosynthesis and carotenoids as accessory pigments. The characteristic pigment profile of the Eustigmatophyceae was one of the evidences which contributed to the segregation of this class of organisms from the Xanthophyceae. Recent findings indicate that eustigmatophyte carotenoids are interesting compounds with laboratory-scale proven health-promoting effects but only a few studies dedicate to the characterization of the pigment profile of the class. In the present work, extracts of 27 eustigmatophyte strains held at the Coimbra Collection of Algae (ACOI) were prepared and analyzed by HPLC-DAD. Results showed a typical eustigmatophyte pigment profile present in all strains, namely, chlorophyll a, violaxanthin, vaucheriaxanthin diester, β,β-carotene, and other minor carotenoids. Violaxanthin was the most abundant carotenoid, achieving nearly half the total pigment content in Monodopsis unipapilla ACOI 2938, and also in two studied Vischeria strains, representing ca. 70% of carotenoids in both strains. The second major carotenoid was β,β-carotene, with the highest production of this commercially important carotenoid detected in Pseudostaurastrum sp. ACOI 3413 (24% of total pigments. Characiopsis cf. saccata ACOI 481 and Characiopsis minuta ACOI 2423 had the highest amount of vaucheriaxanthin diester (19% of total pigments). To our best knowledge, this represents the first report where the pigment profile of eustigmatophytes with stipitate Characiopsis-like morphology is characterized. The results reveal the high potential use of eustigmatophytes as sources of naturally derived carotenoids.

Field-grown grapevines are often exposed to multiple environmental stresses, which challenges wine-growers to develop sustainable measures to sustain vine growth, yield, and quality. Under field conditions this task is demanding, due to differences in the magnitudes of stresses and associated plant responses. In this study we explored the hypothesis that kaolin-particle film application improves grapevine photoprotection through the regulation of xanthophyll cycle genes, limiting the thermal dissipation of excess energy under harsh environmental conditions. Hence, we selected two grapevine varieties, Touriga-Nacional (TN) and Touriga-Franca (TF), grown in the Douro Demarcated Region, and evaluated changes in light dissipation mechanisms, xanthophyll cycle components, and the expression of xanthophyll cycle genes during the 2017 summer season. The results showed that, from veraison to ripening, kaolin triggered the up-regulation of violaxanthin de-epoxidase (VvVDE1) and zeaxanthin epoxidase (VvZEP1) genes, indicating optimised regulation of the xanthophyll cycle. Kaolin treatment also decreased chlorophyll (Chla, Chlb, Chl(a+b)) and carotenoid (Car) accumulation under increasing summer stress conditions in both varieties and lowered the non-photochemical quenching (NPQ) of grapevines on ripening, suggesting a long-term response to summer stress. In addition, kaolin-treated grapevines showed increased Chla/Chlb and lower Chl(a+b)/Car ratios, displaying some features of high light adapted leaves. Overall, this study suggests that kaolin application enabled grapevines to benefit from fluctuating periods of summer stress by managing chlorophyll and carotenoid content and limiting down-regulation of both photochemistry and photoinhibition processes. Under Mediterranean field conditions, kaolin application can be considered an efficient method of minimising summer stress impact on grapevines., This work was supported by National Funds by FCT - Portuguese Foundation for Science and Technology, under the project UIDB/04033/2020, the Clim4Vitis project – “Climate change impact mitigation for European viticulture: knowledge transfer for an integrated approach”, funded by the European Union's Horizon 2020 Research and Innovation programme, under grant agreement no. 810176, and by the project I&D&I AgriFood XXI, operation NORTE-01-0145-FEDER-000041, co-funded by the European Regional Development Fund (FEDER) through NORTE 2020 (Programa Operacional Regional do Norte, 2014/2020). Sara Bernardo acknowledges the financial support provided by the FCT-Portuguese Foundation for Science and Technology (PD/BD/128273/2017) under the Doctoral Programme “Agricultural Production Chains – from Fork to Farm”.

Light, or visible radiation, serves as a source of energy for photosynthesis of plants and most algae. In addition, light and ultraviolet radiation (UV-A and UV-B) act as a biological signal, triggering several cellular processes that are mediated by photoreceptors. The aim of this study was to evaluate the physiological and biochemical responses of Osmundea pinnatifida driven by different radiations through putative photoreceptors. For this, O. pinnatifida was grown under different radiation treatments composed by high intensity of light emitted by a low pressure sodium lamp (SOX), aiming to saturate photosynthesis, which was supplemented by low intensities of visible (red, green and blue) and ultraviolet radiation (UV-A and UV-B), in order to activate photoreceptors. Growth rates, photosynthesis, antioxidant activity, polyphenols, soluble proteins, phycobiliproteins, mycosporine-like amino acids (MAAs) and carotenoids were evaluated during the experiment. Complementary UV-A radiation positively influenced growth rates after 15 days of experiment, although the presence of a peak of blue light in this treatment can also have contributed. UV-B radiation increased the concentration of zeaxanthin and chlorophyll a. The blue light caused the accumulation of chlorophyll a, violaxanthin, phycoerythrin and polyphenols on different days of the experiment. Phycoerythrin also increased under green and red light conditions. Our results showed that some compounds can be modulated by different radiation, and the involvement of photoreceptors is suggested. In red algae, photoreceptors sensitive to red, green and blue light have been identified, however little is known about UV photoreceptors. The presence of photoreceptors sensitive to UV radiation in O. pinnatifida is discussed.

Chickpea is sensitive to heat stress and generally experiences high temperature during reproductive phase. Therefore, terminal heat stress is key constraints for its production. Exogenous application of bioregulators is an option to improve heat tolerance and yield. Therefore, an experiment was carried out using two contrasting Desi type (JG 14) and Kabuli type (Pusa 1108) chickpea varieties with an objective to interlink bioregulators induced heat tolerance with zeaxanthin pigments cycle. Experiment was comprising two main treatments i.e. Normal (18th November) and late (20th December) sown conditions for exposing the crop to high temperature and four sub-treatments (Foliar application of bioregulators viz ABA (10 ppm), BA (40 ppm), SA (100 ppm), thiourea (1000 ppm) and water spray (as control) prior to flowering. Under high temperature condition, reduction in leaf photosynthesis, photosynthetic pigments, chlorophyll fluorescence parameters, yield and its attributes was recorded. Foliar application of bioregulators augmented the grain yield by improving the photosynthetic rate (PN), photosynthetic pigments including zeaxanthin, maximum quantum yield of PS II (Fv/Fm), non-photochemical quenching, heat tolerance index, yield attributes and reducing the level of violaxanthin and heat susceptibility index over the control (water) under late sown condition which in turn suggested that bioregulators induced the heat tolerance by activating operation of zeaxanthin pigments cycle in chickpea. Moreover, Kabuli chickpea variety responded better to bioregulators than Desi type particularly under heat stress condition.

Solar‐induced fluorescence (SIF) is highly relevant in mapping photosynthesis from remote‐sensing platforms. This requires linking SIF to photosynthesis and understanding the role of nonphotochemical quenching (NPQ) mechanisms under field conditions. Hence, active and passive fluorescence were measured in Arabidopsis with altered NPQ in outdoor conditions. Plants with mutations in either violaxanthin de‐epoxidase (npq1) or PsbS protein (npq4) exhibited reduced NPQ capacity. Parallel measurements of NPQ, photosystem II efficiency, SIF and spectral reflectance (ρ) were conducted diurnally on one sunny summer day and two consecutive days during a simulated cold spell. Results showed that both npq mutants exhibited higher levels of SIF compared to wild‐type plants. Changes in reflectance were related to changes in the violaxanthin–antheraxanthin–zeaxanthin cycle and not to PsbS‐mediated conformational changes. When plants were exposed to cold temperatures, rapid onset of photoinhibition strongly quenched SIF in all lines. Using well‐characterized Arabidopsis npq mutants, we showed for the first time the quantitative link between SIF, photosynthetic efficiency, NPQ components and leaf reflectance. We discuss the functional potential and limitations of SIF and reflectance measurements for estimating photosynthetic efficiency and NPQ in the field.

Xanthophylls in light harvesting complexes perform a number of functions ranging from structural support to light-harvesting and photoprotection. In the major light harvesting complex of photosystem II in plants (LHCII), the innermost xanthophyll binding pockets are occupied by lutein molecules. The conservation of these sites within the LHC protein family suggests their importance in LHCII functionality. In the present work, we induced the photoprotective switch in LHCII isolated from the Arabidopsis mutant npq1lut2, where the lutein molecules are exchanged with violaxanthin. Despite the differences in the energetics of the pigments and the impairment of chlorophyll fluorescence quenching in vivo, we show that isolated complexes containing violaxanthin are still able to induce the quenching switch to a similar extent to wild type LHCII monomers. Moreover, the same spectroscopic changes take place, which suggest the involvement of the terminal emitter site (L1) in energy dissipation in both complexes. These results indicate the robust nature of the L1 xanthophyll binding domain in LHCII, where protein structural cues are the major determinant of the function of the bound carotenoid.

Chickpea being a winter season crop often experiences heat stress during reproductive phase. For chickpea production, terminal heat stress is one of the major constraints. Plants have built up numerous mechanisms to combat the heat stress. We considered the photosynthetic pigments for heat tolerance. Therefore, in order to investigate the heat tolerance in relation to photosynthetic pigments, a field trial was carried out having 4 contrasting genotypes namely BG 240 and JG 14 (relatively heat tolerant), SBD 377 (moderately tolerant) and ICC 1882 (relatively heat sensitive). Heat stress was imposed by altering the sowing date i.e. normal (18th November) and late sown (18th December). Under delayed sown condition, heat stress was faced by crop starting from flowering stage to crop maturity. Under heat stress condition, heat tolerant genotypes BG 240 and JG 14 maintained higher level of membrane stability, RWC (%), osmolytes, dry matter partitioning, grain yield, heat tolerance index and had higher values of zeaxanthin, quantum yield of PS II (Fv/Fm ratio), non-photochemical quenching (NPQ), photosynthetic rate, level of photosynthetic pigments (chlorophylls and carotenoids) and lower level of violaxanthin, and lipid peroxidation as compared to heat sensitive one (ICC 1882). In addition to this, Fv/Fm ratio and NPQ exhibited positive relationship with heat tolerance which suggested the involvement of xanthophyll cycle pigments in chickpea heat tolerance.

Optimizing the photoprotection of different leaves as a whole is important for plants to adapt to fluctuations in ambient light conditions. However, the molecular basis of this leaf-to-leaf communication is poorly understood. Here, we used a range of techniques, including grafting, chlorophyll fluorescence, revers transcription quantitative PCR, immunoblotting, chromatin immunoprecipitation, and electrophoretic mobility shift assays, to explore the complexities of leaf-to-leaf light signal transmission and activation of the photoprotective response to light fluctuation in tomato (Solanum lycopersicum). We established that light perception in the top leaves attenuated the photoinhibition of both PSII and PSI by triggering photoprotection pathways in the bottom leaves. Local light promoted the accumulation and movement of LONG HYPOCOTYL5 from the sunlit local leaves to the systemic leaves, priming the photoprotective response of the latter to light fluctuation. By directly activating the transcription of PROTON GRADIENT REGULATION5 and VIOLAXANTHIN DE-EPOXIDASE, LONG HYPOCOTYL5 induced cyclic electron flow, the xanthophyll cycle, and energy-dependent quenching. Our findings reveal a systemic signaling pathway and provide insight into an elaborate regulatory network, demonstrating a pre-emptive advantage in terms of the activation of photoprotection and, hence, the ability to survive in a fluctuating light environment.

Dynamics of chlorophyll a fluorescence changes (including JIP test parameters) and photosynthetic pigments were studied in oat (Avena sativa L.) leaves of late summer sowing during autumnal cooling. As the average daily temperature decreased from 8 to 2°С, the chlorophyll content decreased by 40%, while the chlorophyll a/b ratio ranged between 2.2 and 2.4. The total pool of carotenoids and the relative content of β‑carotene, lutein, neoxanthin, and pigments of the violaxanthin cycle changed insignificantly. The relative number of QB-nonreducing reaction centers (RC) of photosystem II (PSII) and the quantum yield of energy-dependent component qE of nonphotochemical quenching in the PSII antenna (φNPQ parameter) were found to increase during the seasonal cooling. The values of Fv/Fm parameter characterizing the maximal photochemical efficiency of PSII complexes were 0.72–0.76. At temperatures around 0°C, Fv/Fm decreased to 0.62. At the same time, the nonregulated (light-independent) thermal dissipation (photoinhibitory component qI of nonphotochemical quenching) was enhanced, which was manifested in the increase in parameters φDo and DI0/RC. It is concluded that, under seasonal cooling, the PSII reaction centers partly lose photochemical activity and start functioning as dissipative centers of excitation energy. In a significant part of inactivated RC, the loss of QA-reduction was reversible, which implies the formation of photoinactivated forms of PSII complexes stabilized at low temperatures. The decrease in daily average temperatures to −3°С with nighttime low temperatures from −7 to −8°С led to plant death within 3 days.

The present study investigated how different storage conditions affect antioxidant levels in three scab-resistant and powdery mildew-tolerant apple cultivars. The contents of carotenoids, along with free and glycosylated phenolic acids, were quantified in harvested apples and apples stored in boxes with controlled atmospheric conditions (1.2% O2 and 2.2% CO2) and temperature (1 °C), boxes with regulated temperature (1 °C) and storage rooms with a temperature between 1 and 4 °C. All three cultivars demonstrated a continuous decrease in total carotenoid content (sum of s-carotene, lutein, neoxanthin, violaxanthin, zeaxanthin and antheraxanthin) during the storage period. However, apples stored under controlled atmospheric conditions showed significantly higher carotenoid levels than apples stored under other approaches over 3 and 5 months of storage. Although the storage approach clearly affected carotenoid levels across the three tested cultivars, the ratio between the levels of individual carotenoids was neither affected by storage approach nor cultivar. Free phenolic acid contents were found to be cultivar specific; whereas, all three cultivars demonstrated similar glycosylated phenolic acid contents. Chlorogenic acid was the most common free phenolic acid in all three cultivars, while protocatechuic and caffeic acid were the dominant glycosylated phenolic acids identified from the cultivars. Interestingly, stored apples showed higher free phenolic acid levels than harvested apples; while, both harvested and stored apples showed similar concentrations of glycosylated phenolic acids. As such, none of the three tested storage conditions had a pronounced effect on either group of phenolic acids (total content or concentrations of individual compounds).

The accumulation of carotenoids (carotenogenesis) in the cells of Haematococcus pluvialis (strain IBCE H-17) upon cultivation in nutrient media deficient in nutrient elements (N and K + P) is shown. K and P deficiency in nutrient medium led to accumulation of astaxanthin, β-carotene, anteraxanthin, zeaxanthin and violaxanthin after 18 days of cultivation, while the amount of neoxanthin and lutein was at the control level. Carotenogenesis under such conditions was accompanied by high levels of chlorophyll (a + b). The use of nutrient medium deficient in N led to the accumulation of astaxanthin and, to a lesser extent, β-carotene in haematococcus cells, while lutein, anteraxanthin, zeaxanthin, violaxanthin and neoxanthin levels were lower or the same as control. N deficiency caused the decrease in amount of chlorophyll (a + b). Nitrogen deficiency was 4.3 times more effective than control medium and 1.8 times more effective than combined potassium and phosphorus deficiency in promoting astaxanthin accumulation.

Microbial production of carotenoids has mainly focused towards a few products, such as β-carotene, lycopene and astaxanthin. However, other less explored carotenoids, like violaxanthin, have also shown unique properties and promissory applications. Violaxanthin is a plant-derived epoxidated carotenoid with strong antioxidant activity and a key precursor of valuable compounds, such as fucoxanthin and β-damascenone. In this study, we report for the first time the heterologous production of epoxycarotenoids in yeast. We engineered the yeast Saccharomyces cerevisiae following multi-level strategies for the efficient accumulation of violaxanthin. Starting from a β-carotenogenic yeast strain, we first evaluated the performance of several β-carotene hydroxylases (CrtZ), and zeaxanthin epoxidases (ZEP) from different species, together with their respective N-terminal truncated variants. The combined expression of CrtZ from Pantoea ananatis and truncated ZEP of Haematococcus lacustris showed the best performance and led to a yield of 1.6 mg/gDCW of violaxanthin. Further improvement of the epoxidase activity was achieved by promoting the transfer of reducing equivalents to ZEP by expressing several redox partner systems. The co-expression of the plant truncated ferredoxin-3, and truncated root ferredoxin oxidoreductase-1 resulted in a 2.2-fold increase in violaxanthin yield (3.2 mg/gDCW). Finally, increasing gene copy number of carotenogenic genes enabled reaching a final production of 7.3 mg/gDCW in shake flask cultures and batch bioreactors, which is the highest yield of microbially produced violaxanthin reported to date.

The violaxanthin (V)-antheraxanthin (A)-zeaxanthin (Z) (VAZ) cycle was deemed a non-second-scale process of photoprotection in higher plants and microalgae, but the validity of this view has not been confirmed. To test this view, we explored responses of the VAZ cycle and the relationship between the VAZ cycle and non-photochemical quenching (NPQ) under highlight at second and minute scales in Heterosigma akashiwo and Platymonas sp. Both A and Z were generated in H. akashiwo during 15 s of light exposure, whereas only A rapidly accumulated within 15 s of exposure in Platymonas sp. The above results, together with a time-dependent sigmoidal relationship between the VAZ cycle (de-epoxidation state, A/Chl a, and Z/Chl a) and NPQ, proved that the VAZ cycle was a second-scale process related to NPQ. In addition, we found that not all NPQ was dependent on the VAZ cycle and suggested that NPQ model should be carefully modified due to the species-specific proportions of de-epoxidation-dependent NPQ.

Non-photochemical quenching (NPQ) is one of the most important photo-protection mechanisms in brown macroalgae. Global warming and ocean acidification are predicted to impact physiological characteristics of marine algae. However, little is known about the effects of co-occurrence of the elevation of pCO2 and temperature on photochemical capacity, especially regarding photoprotective mechanisms in brown macroalgae. Here, we studied the separate and combined effects of increases in pCO2 and temperature on the photochemical characteristics and growth performance in sporophytes of the brown macroalga Saccharina japonica. The results showed that the NPQ of S. japonica is mainly dependent on the xanthophyll cycle (XC) which appears to be related only to the activation of the enzyme violaxanthin de-epoxidase (VDE), and the transthylakoid proton gradient (ΔpH) could not induce NPQ alone. The elevation of pCO2 reduced NPQ value of S. japonica under high-temperature stress. After 60-day cultivation under the ambient and elevated pCO2 (400 and 1000 μatm), we further found that the elevation of pCO2 promoted growth and increased the photosynthetic performance of all three cultivar strains of S. japonica that have been traditionally cultured for many years in China.

Recombinant Escherichia coli cells, which express the higher-plant CaZEP (zeaxanthin epoxidase) and AtCYO1 (molecular chaperone) genes and an NADPH regeneration system in addition to the IDI, crtE, crtB, crtI, crtY, and crtZ genes for zeaxanthin biosynthesis, were cultured in 2 × YT medium for the efficient production of antheraxanthin and violaxanthin. The carotenoids accumulated in the cells were purified by two-phase partition, silica gel column chromatography, and preparative silica gel HPLC. Mass spectrometry and nuclear magnetic resonance analyses of these carotenoids revealed that a novel carotenoid, determined to be (3R,6′S)-3,6′-dihydroxy-β,e-caroten-3′-one and named as 6′-hydroxy-3′-didehydrolutein, was produced in addition to zeaxanthin, antheraxanthin and violaxanthin. It is likely that the new carotenoid was generated from an unstable epoxy-carotenoid antheraxanthin that was synthesized from zeaxanthin with CaZEP. Its antioxidant activities were also examined.

Heatwaves are becoming more frequent with climate warming and can impact tree growth and reproduction. Eucalyptus parramattensis can cope with an extreme heatwave in the field via transpiratory cooling and enhanced leaf thermal tolerance that protected foliar tissues from photo-inhibition and photo-oxidation during natural midday irradiance. Here, we explored whether changes in foliar carotenoids and/or the xanthophyll cycle state can facilitate leaf acclimation to long-term warming and/or an extreme heatwave event. We found that leaves had similar carotenoid levels when grown for one year under ambient and experimental long-term warming (+ 3 °C) conditions in whole tree chambers. Exposure to a 4-day heatwave (> 43 °C) significantly altered the xanthophyll de-epoxidation state of carotenoids revealing one mechanism by which trees could minimise foliar photo-oxidative damage. The levels of zeaxanthin were significantly higher in both young and old leaves during the heatwave, revealing that violaxanthin de-epoxidation and perhaps de novo zeaxanthin synthesis contributed to enhancement of the xanthophyll cycle state. In a future climate of long-term warming and increased heatwave events, leaves of E. parramattensis will be able to utilise biochemical strategies to alter the xanthophyll cycle state and cope with extreme temperatures under natural solar irradiation.

Carotenoids are a class of natural product compound that are currently being used as colouring agents and widely used in food industry. Carotenoids are bioactive pigments obtained mainly from plants through dietary intake. They possess good features in terms of dietary supplement, food colourant, and polymer stabiliser. The presence of 4 main carotenoids, which are β-carotene, zeaxanthin, lutein, and violaxanthin, were determined in 4 classes of allelopathic plant groups namely trees, ferns, grasses and herbaceous plants. This research aims to explore the carotenoid’s content and composition in 11 allelopathic species by HPLC analysis. A. auriculiformis (tree) was found to have the highest total carotenoid concentration (146.36 µg/g DW) that was substantially higher than all other species tested whereas the lowest total carotenoid concentration was found in S. palustris (fern) (3.76 µg/g DW). Lutein and β-carotene were detected highest in A. auriculiformis (tree), with 1024 ± 25.5 µg/g DW and 37.55 ± 3.16 µg/g DW, respectively. Violaxanthin and zeaxanthin were found substantially highest in M. cajuputi (tree) (5.02 ± 0.5 µg/g DW) and S. palustris (fern) (5.88 ± 0.19µg/g DW), respectively.

We present the results of experiments on the influence of feeding the marine bivalve mollusk Cerastoderma glaucum (Bruguiere, 1789) with a monoculture of the green algae Tetraselmis viridis on the composition of carotenoids in its tissues. The carotenoid repertoire of C. glaucum was found to include β-carotene, alloxanthin, lutein, diadinoxanthin, neoxanthin, fucoxanthin, heteroxanthin, mactraxanthin and four carotenoid esters, with mactraxanthin being specific to C. glaucum. In T. viridis, carotenoids were represented by β, e -carotene, lutein, zeaxanthin, violaxanthin, neoxanthin. A 6-day microalgal mono-diet led to an increase in the total carotenoid content in mollusk tissues, mainly at the expense of pigments characteristic of T. viridis. The process was accompanied by a decrease in the concentration of other plant carotenoids and mactraxanthin isomerization. Despite the changes in the accumulation and transformation of carotenoids, the experiment led to a 20% death rate among individuals, indicating an adverse effect of a monotonous diet on mollusks.

Chlorella gloriosa (Chlorellaceae, Trebouxiophyceae) was isolated from seawater off the coast of the Dokdo Islands in Korea. An axenic culture was established using the streak-plate method on f/2 agar media supplemented with antibiotics, allowing identification of the isolate by morphological, molecular, and physiological analyses. The morphological characteristics observed by light and electron microscopy revealed typical morphologies of C. gloriosa species. The molecular phylogenetic inference drawn from the small-subunit 18S rRNA sequence verified that the microalgal strain belongs to C. gloriosa. Additionally, gas chromatography-mass spectrometry analysis showed that the isolate was rich in nutritionally important omega-3 and -6 polyunsaturated fatty acids and high-performance liquid chromatography analysis revealed that the high-value antioxidants lutein and violaxanthin were biosynthesized as accessory pigments by this microalga, with arabinose, galactose, and glucose as the major monosaccharides. Therefore, in this study, a Korean marine C. gloriosa species was discovered, characterized, and described, and subsequently added to the national culture collection.

Cera pepper ( Capsicum pubescens ) is an exotic fruit considered as a rich source of nutraceuticals with known benefits for human health and also an economic resource for local producers in Mexico. The present investigation reports on the in vitro and in situ antifungal activity of the essential oil from Mentha piperita and its two major volatiles (menthol and menthone) against Fusarium sambucinum , which is a causal agent of soft rot in cera pepper. The application of these components in pepper fruits previously infected with F. sambucinum caused a significant delay (p < 0.05) in the emergence of soft rot symptoms. This effect was reflected in the maintenance of pH and fruit firmness during a period of 10 days. The nutrimental content of the fruits (protein, fiber, fat and other proximate parameters) was conserved in the same period of time. The nutraceutical content of these fruits was estimated by the quantification of seven carotenoids (violaxanthin, cis-violaxanthin, luteoxanthin, antheraxanthin, lutein, zeaxanthin and beta-carotene), ascorbic acid and capsaicinoids (capsaicin and dihydrocapsaicin). According to our results, the essential oil from M. piperita and its major volatiles exerted a preservative effect on these metabolites. Our findings demonstrated that the essential oil of M. Piperita and its major volatiles represent an ecological alternative for the control of fusariosis caused by F. sambucinum in cera peppers under postharvest conditions.

OsVDE, a lipocalin-like protein in chloroplasts, negatively regulated the ABA biosynthesis and stomatal closure under salt stress in rice seedlings. Violaxanthin de-epoxidase (VDE) is a key enzyme of xanthophyll cycle. It plays a critical role in abscisic acid (ABA) biosynthesis, growth and stress responses in plants. Although functions of several VDE genes have been characterized, it is largely unknown whether OsVDE regulates the ABA biosynthesis and salt stress tolerance in rice. In this study, we generated the OsVDE overexpressing and CRISPR-Cas9-mediated gene-editing transgenic lines, and identified that the gene-editing mutant lines showed the dwarfism, shorter panicle and lower seed-setting rate than the wild type whereas the overexpression lines did not exhibit the difference from the wild type. In addition, the gene-editing transgenic lines were hypersensitive to exogenous ABA during germination. Under salt stress, the gene-editing transgenic seedlings had a higher ABA level, higher stomatal closure percentage and higher survival rate than the wild type. The qRT-PCR analysis confirmed that OsVDE negatively regulated the OsNECD2/4/5 expressions, ABA biosynthesis and salt stress tolerance in rice seedlings. These results provide new evidence that VDE plays an essential role in ABA biosynthesis and salt stress tolerance in plants.

To determine the role of α- and γ-tocopherol (TC), this study compared the response to salt stress (200 mM NaCl) in wild type (WT) Arabidopsis thaliana (L.) Heynh. And its two mutants: (1) totally TC-deficient vte1, (2) vte4 accumulating γ-TC instead of α-TC, and (3) tmt transgenic line overaccumulating α-TC. Raman spectra revealed that salt-exposed α-TC accumulating plants were more flexible in regulating chlorophyll, carotenoid and polysaccharide levels than TC deficient mutants, while the plants overaccumulating γ-TC had the lowest levels of these biocompounds. Tocopherol composition and NaCl concentration affected xanthophyll cycle by changing the rate of violaxanthin de-epoxidation and zeaxanthin formation. NaCl treated plants with altered TC composition accumulated less oligosaccharides than WT plants. α-TC deficient plants increased their oligosaccharide levels and reduced maltose amount, while excessive accumulation of α-TC corresponded with enhanced amounts of maltose. Salt-stressed TC-deficient mutants and tmt transgenic line exhibited greater proline levels than WT plants, lower chlorogenic acid levels, and lower activity of catalase and peroxidases. α-TC accumulating plants produced more methylated proline- and glycine- betaines, and showed greater activity of superoxide dismutase than γ-TC deficient plants. Under salt stress, α-TC demonstrated a stronger regulatory effect on carbon- and nitrogen-related metabolites reorganization and modulation of antioxidant patterns than γ-TC. This suggested different links of α- and γ-TCs with various metabolic pathways via various functions and metabolic loops.

Main conclusion The compatible solute sucrose reduces the efficiency of the enzymatic de-epoxidation of violaxanthin, probably by a direct effect on the protein parts of violaxanthin de-epoxidase which protrude from the lipid phase of the thylakoid membrane. The present study investigates the influence of the compatible solute sucrose on the violaxanthin cycle of higher plants in intact thylakoids and in in vitro enzyme assays with the isolated enzyme violaxanthin de-epoxidase at temperatures of 30 and 10 °C, respectively. In addition, the influence of sucrose on the lipid organization of thylakoid membranes and the MGDG phase in the in vitro assays is determined. The results show that sucrose leads to a pronounced inhibition of violaxanthin de-epoxidation both in intact thylakoid membranes and the enzyme assays. In general, the inhibition is similar at 30 and 10 °C. With respect to the lipid organization only minor changes can be seen in thylakoid membranes at 30 °C in the presence of sucrose. However, sucrose seems to stabilize the thylakoid membranes at lower temperatures and at 10 °C a comparable membrane organization to that at 30 °C can be observed, whereas control thylakoids show a significantly different membrane organization at the lower temperature. The MGDG phase in the in vitro assays is not substantially affected by the presence of sucrose or by changes of the temperature. We conclude that the presence of sucrose and the increased viscosity of the reaction buffers stabilize the protein part of the enzyme violaxanthin de-epoxidase, thereby decreasing the dynamic interactions between the catalytic site and the substrate violaxanthin. This indicates that sucrose interacts with those parts of the enzyme which are accessible at the membrane surface of the lipid phase of the thylakoid membrane or the MGDG phase of the in vitro enzyme assays.

In this study, astaxanthin yield of Phaffia rhodozyma PR106 increased significantly under titanium dioxide (TiO2) stress, and the yield of lycopene and β-carotene also increased significantly, as well as the yield of violaxanthin and lutein significantly decreased; in addition, TiO2 stress promoted cell division and changed cell morphology of PR106. Then, the mechanism of increasing astaxanthin yield was studied by transcriptomics and related metabolic regulation. The results showed that astaxanthin accumulation in PR106 was not directly related to mRNA transcription and post-translational modifications regulation under TiO2 stress; TiO2 stress accelerated glucose uptake of yeast, promoted reuse of ethanol, and increased the formation of acetyl-CoA and ATP. The more carbon flux was shifted to astaxanthin synthesis pathway and weakened carotenoids accumulation in astaxanthin branch pathway to improve the astaxanthin production of PR106. The metabolism regulation of ROS could continue in the PR106 strain.

Thioredoxin-like protein CDSP32 (Trx CDSP32), a thioredoxin-like (Trx-like) protein located in the chloroplast, can regulate photosynthesis and the redox state of plants under stress. In order to examine the role of Trx CDSP32 in the photosynthetic apparatus of plants exposed to cadmium (Cd), the effects of Trx CDSP32 on photosynthetic function and photoprotection in tobacco leaves under Cd exposure were studied using a proteomics approach with wild-type (WT) and Trx CDSP32 overexpression (OE) tobacco plants. Cd exposure reduced stomatal conductance, blocked PSII photosynthetic electron transport, and inhibited carbon assimilation. Increased water use efficiency (WUE), cyclic electron flow (CEF) of the proton gradient regulation 5 pathway (PGR5-CEF), and regulated energy dissipation [Y(NPQ)] are important mechanisms of Cd adaptation. However, CEF of the NAD(P)H dehydrogenase pathway (NDH-CEF) was inhibited by Cd exposure. Relative to control conditions, the expression levels of violaxanthin de-epoxidase (VDE) and photosystem II 22 kDa protein (PsbS) in OE leaves were significantly increased under Cd exposure, but those in WT leaves did not change significantly. Moreover, the expression of zeaxanthin epoxidase (ZE) under Cd exposure was significantly higher than that in WT leaves. Thus, Trx CDSP32 increased Y(NPQ) and alleviated PSII photoinhibition under Cd exposure. Trx CDSP32 not only increased PGR5-like protein 1A and 1B expression, but also alleviated the down-regulation of NAD(P)H-quinone oxidoreductase subunits induced by Cd exposure. Thus, Trx CDSP32 promotes CEF in Cd-exposed tobacco leaves. Thus, Trx CDSP32 alleviates the Cd-induced photoinhibition in tobacco leaves by regulating two photoprotective mechanisms: CEF and xanthophyll cycle-dependent energy dissipation.

RATIONALE Liquid chromatography/photodiode array atmospheric pressure chemical ionization mass spectrometry (LC/PDA-APCI-MS) is used for the analysis of various carotenoid pigments in plants. Among them, it is difficult to distinguish between the isomeric violaxanthin/neoxanthin esters. METHODS The yellow pigments of tomato petals were extracted with acetone, and the extracts were kept at -30°C to allow the contaminating triacylglycerols to settle out physically. The supernatants were analyzed using LC/PDA-APCI-MS with a high-resolution orbitrap mass spectrometer for their exact masses. The expected carotenoid esters were calculated with the combination of carotenoids and fatty acids, and they were matched with the experimental exact masses. The fatty acid structures in the carotenoid esters were also identified using collision-induced dissociation (CID) tandem mass spectrometry (MS/MS). The isomeric violaxanthin/neoxanthin esters were distinguished using CID MS/MS from their in-source dehydrated product ions as pseudoprecursor ions. RESULTS The in-source dehydrated ions [M - H2 O + H]+ of neoxanthin diesters predominated over their protonated molecules [M + H]+ in LC/MS. By contrast, the protonated molecules of violaxanthin diesters predominated. The 92 u loss product ions [M - H2 O - C7 H8 + H]+ were observed from the dehydrated violaxanthin diesters, but they were not generated from the dehydrated neoxanthin diesters in the CID MS/MS of their dehydrated pseudoprecursor ion [M - H2 O + H]+ . CONCLUSIONS The allene allyl carbocation in neoxanthin diesters was generated from dehydration after preferential protonation at the hydroxy group. The epoxide group of violaxanthin diesters opens easily after protonation; however, the dehydration did not proceed at this stage. The 92 u loss of C7 H8 was explained by an intramolecular [2 + 2] cycloaddition, which proceeded preferentially in dehydrated violaxanthin diesters because the carbocations in the dehydrated species were conjugated to the polyene and those double bonds were depolarized during CID MS/MS. Therefore, the isomeric neoxanthin/violaxanthin diesters were distinguished using LC/PDA-APCI-MS and MS/MS. This method was a practical and useful method of profiling the carotenoid esters of the yellow petals.

Simple Summary Photosynthetic organisms, such as land plants, evolved to utilize available light and to use its energy to assimilate carbon dioxide and produce carbohydrates. However, the light intensity often exceeds the ability of plants to successfully utilize absorbed energy, thus inducing stress, manifested by an increased radical concentration inside plant cells and disruption of the inner structures, and consequently decreased plant yield. Plants solve this problem by using a mechanism termed non-photochemical quenching, by which they can dissipate the energy not used in photosynthesis. Modern agriculture, however, also involves indoor plant farming. For indoor plant farming LED-based lighting systems, with non-saturating light intensities, are suitable based on their restricted energy consumption. However, the composition of applied light should first be optimized to maximize its utilization. Our study examined the influence of monochromatic LEDs (red, green, and blue) on the photoprotective and photosynthetic properties of tomato plants. We indicate that monochromatic green light could be considered an important component of lighting systems to alleviate energy dissipation, while blue light enhances photosynthetic efficiency. Our study not only proves the crucial importance of spectrum optimization but also provides evidence that different light wavelengths modify photosynthetic and photoprotective properties. Abstract Photosynthetic pigments of plants capture light as a source of energy for photosynthesis. However, the amount of energy absorbed often exceeds its utilization, thus causing damage to the photosynthetic apparatus. Plants possess several mechanisms to minimize such risks, including non-photochemical quenching (NPQ), which allows them to dissipate excess excitation energy in the form of harmless heat. However, under non-stressful conditions in indoor farming, it would be favorable to restrict the NPQ activity and increase plant photosynthetic performance by optimizing the light spectrum. Towards this goal, we investigated the dynamics of NPQ, photosynthetic properties, and antioxidant activity in the leaves of tomato plants grown under different light qualities: monochromatic red (R), green (G), or blue (B) light (L) at 80 µmol m−2 s−1 and R:G:B = 1:1:1 (referred to as the white light (WL)) at 120 µmol m−2 s−1. The results confirm that monochromatic BL increased the quantum efficiency of PSII and photosynthetic pigments accumulation. The RL and BL treatments enhanced the NPQ amplitude and showed negative effects on antioxidant enzyme activity. In contrast, plants grown solely under GL or WL presented a lower amplitude of NPQ due to the reduced accumulation of NPQ-related proteins, photosystem II (PSII) subunit S (PsbS), PROTON GRADIENT REGULATION-LIKE1 (PGRL1), cytochrome b6f subunit f (cytf) and violaxanthin de-epoxidase (VDE). Additionally, we noticed that plants grown under GL or RL presented an increased rate of lipid peroxidation. Overall, our results indicate the potential role of GL in lowering the NPQ amplitude, while the role of BL in the RGB spectrum is to ensure photosynthetic performance and photoprotective properties.

Carotenoids are naturally synthesized in some species of bacteria, archaea, and fungi (including yeasts) as well as all photosynthetic organisms. Escherichia coli has been the most popular bacterial host for the heterologous production of a variety of carotenoids, including even xanthophylls unique to photosynthetic eukaryotes such as lutein, antheraxanthin, and violaxanthin. However, conversion efficiency of these epoxy-xanthophylls (antheraxanthin and violaxanthin) from zeaxanthin remained substantially low. We here examined several factors affecting their productivity in E. coli. Two sorts of plasmids were introduced into the bacterial host, i.e., a plasmid to produce zeaxanthin due to the presence of the Pantoea ananatis crtE, crtB, crtI, crtY, and crtZ genes in addition to the Haematococcus pluvialis IDI gene, and one containing each of zeaxanthin epoxidase (ZEP) genes originated from nine photosynthetic eukaryotes. It was consequently found that paprika (Capsicum annuum) ZEP (CaZEP) showed the highest conversion activity. Next, using the CaZEP gene, we performed optimization experiments in relation to E. coli strains as the production hosts, expression vectors, and ribosome-binding site (RBS) sequences. As a result, the highest productivity of violaxanthin (231 μg/g dry weight) was observed, when the pUC18 vector was used with CaZEP preceded by a RBS sequence of score 5000 in strain JM101(DE3).

Chlorophyll triplet excited states are byproducts of photosynthetic processes that can indirectly harm biological membranes by forming highly reactive oxygen species. A crucial photoprotective mechanism evolved by plants to counter this threat involves the triplet energy transfer from chlorophylls to carotenoid molecules, in which triplet states are not reactive. In the major light-harvesting complex of photosystem II (LHCII), the two central luteins play an important role in the mechanism, but it has been shown that carotenoid triplets are formed even when other carotenoids replace them in their binding sites. In this work, we have investigated carotenoid triplet formation in LHCII isolated from

The variation of flower color of chrysanthemum (Chrysanthemum×morifolium) is extremely rich, and carotenoids, which are mainly stored in the plastid, are important pigments that determine the color of chrysanthemum. However, the genetic regulation of the carotenoid metabolism pathway in this species still remains unclear. In this study, a pink chrysanthemum cultivar, 'Jianliuxiang Pink', and its three bud sport mutants (including white, yellow and red color mutants, 'Jianliuxiang White', 'Jianliuxiang Yellow' and 'Jianliuxiang Red', respectively) were used as experimental materials to analyze the dynamic changes of carotenoid components and plastid ultrastructure at different developmental stages of ray florets. We found that the carotenoid components and plastid ultrastructure of the four color cultivars in the early developmental stage of the chrysanthemum capitulum (S1) were almost identical, and the carotenoids mainly included violaxanthin, lutein and β-carotene, which exist in proplastids and immature chloroplasts. With the development of capitulum, the chloroplasts in 'Jianliuxiang White' and 'Jianliuxiang Pink' were degraded, and the protoplasts did not transform but rather formed vesicles that accumulated trace amounts of carotenoids. The proplastids and chloroplasts in 'Jianliuxiang Yellow' and 'Jianliuxiang Red' were all transformed into chromoplasts and consist of lutein as well as lutein's isomer and derivatives. Using comparative transcriptomics combined with gene expression analysis, we found that CmPg-1, CmPAP10, and CmPAP13, which were involved in chromoplast transformation, CmLCYE, which was involved in carotenoid biosynthesis, and CmCCD4a-2, which was involved in carotenoid degradation, were differentially expressed between four cultivars, and these key genes therefore should affect the accumulation of carotenoids in chrysanthemum. In addition, six transcription factors, CmMYB305, CmMYB29, CmRAD3, CmbZIP61, CmAGL24, CmNAC1, were screened using weighted gene co-expression correlation network analysis (WGCNA) combined with correlative analysis to determine whether they play an important role in carotenoid accumulation by regulating structural genes related to the carotenoid metabolism pathway and plastid development. This study analyzed dynamic changes of carotenoid components and plastid ultrastructure of the four bud mutation cultivars of chrysanthemum and identified structural genes and transcription factors that may be involved in carotenoid accumulation. The above results laid a solid foundation for further analysis of the regulatory mechanism of the carotenoid biosynthesis pathway in chrysanthemum.

Carotenoids are the most important pigments determining the color of C. × morifolium; however, it is still unknown whether the changes of plastid ultrastructure affect carotenoids accumulation. In this study, we compared the change of carotenoid composition, content, and the plastid ultrastructures in the different developmental stages of capitulum among fourteen C. × morifolium cultivars from seven color groups. We found that the carotenoids and plastids detected at the early stage of capitulum development were similar in all cultivars, including violaxanthin, lutein, and β-carotene, which were present in proplastids and immature chloroplasts. Immature chloroplasts were degraded completely, forming loosely broken plastids during the development of the capitulum in white and pink cultivars. Meanwhile, a number of lipid vesicles appeared at proplastids, which resulted in only trace amounts of carotenoid accumulation in these cultivars. For yellow, orange, red, and brown cultivars, a great number of chromoplasts were found, which contained diverse ultrastructures, such as plastoglobules, tubules, and lipid droplets; these chromoplasts were derived from proplastids or chloroplasts. Compared with the early stage of capitulum development, these cultivars accumulated large amounts of carotenoids, primarily including lutein, lutein derivatives, and their isomers. In green cultivars, proplastids and immature chloroplasts were completely transformed into mature chloroplasts. These chloroplasts mainly contained violaxanthin, lutein, β-carotene, and two new components, (9Z)-lutein and (9'Z)-lutein, compared with carotenoid components presented in proplastids and immature chloroplasts. This research will be helpful for understanding the mechanisms of carotenoid metabolism of C. × morifolium. Furthermore, we found that two different chromoplast transformation patterns could be present in the same tissue cell, which contributed to the research on plastid differentiation and development in higher plants.

For winter survival, evergreens not only need cold acclimation (CA), but also photoprotection. This study was conducted to examine how two elevational ecotypes of Rhododendron catawbiense respond to cold and light stresses in a common garden. Seasonal changes in leaf freezing tolerance (LFT), photosystem II efficiency, photosynthesis, carotenoids (xanthophyll cycle pigments, lutein, beta-carotene, neoxanthin) were investigated from summer to winter. Parallel changes in the expression of violaxanthin de-epoxidase (VDE) gene were examined to explore its association with violaxanthin de-epoxidation. High elevation (HE) ecotype attained overall greater LFT while low elevation (LE) ecotype was more responsive to short-days for the first stage of CA. In summer, both ecotypes predominantly employed reversible xanthophyll cycle (RXC) to dissipate excess light energy, but LE used it more robustly. In autumn, only HE exhibited slight damage to the reaction centers (RCs), i.e. photosystem II degradation (PSD). In winter, both ecotypes suspended photosynthesis, were photoinhibited, and employed primarily sustained de-epoxidation (SD) for photoprotection; but LE still used RXC more effectively than HE. Overall, LE employed xanthophyll cycle more robustly in summer or winter. Results suggest that microclimates of R. catawbiense ecotypes inform their differential physiological response to cold and light stresses. VDE expression data indicate violaxanthin de-epoxidation in R. catawbiense may not be solely transcriptionally regulated.

The aim of this study was to evaluate the impact of selected types of LED (light emitting diodes) lighting on the quality of alfalfa sprouts. In the experiment, cold white, warm white and multicolour: (red, green, blue-RGB) LEDs were applied, and dispersed sunlight was used as a control. The product was examined for the yield and the contents of dry matter, total polyphenols, ascorbic acid, chlorophylls, β-carotene, lutein, neoxanthin and violaxanthin. Cotyledons' mass in the whole plant increased under LED illumination and was up to 50% greater for sprouts grown in RGB light compared to those cultivated in dispersed sunlight. The highest chlorophyll and carotenoid pigment contents in cotyledons were observed under RGB LED and cold white treatments. Similarly, RGB LEDs allows one to obtain the product with the highest level of total phenolic compounds. The highest ascorbic acid content was observed in sprouts growing under sunlight, followed by RGB.

Plants have developed multiple self-regulatory mechanisms to efficiently function under varying sunlight conditions. At high light intensities, non-photochemical quenching (NPQ) is activated on a molecular level, safely dissipating an excess excitation as heat. The exact molecular mechanism for NPQ is still under debate, but it is widely agreed that the direct participation of the carotenoid pigments is involved, one of the proposed candidate being the zeaxanthin. In this work, we performed fluorescence measurements of violaxanthin- and zeaxanthin-enriched major light-harvesting complexes (LHCII), in ensemble and at the single pigment-protein complex level, where aggregation is prevented by immobilization of LHCIIs onto a surface. We show that a selective enrichment of LHCII with violaxanthin or zeaxanthin affects neither the ability of LHCII to switch into a dissipative conformation nor the maximal level of induced quenching. However, the kinetics of the fluorescence decrease due to aggregation on the timescale of seconds are different, prompting towards a modulatory effect of zeaxanthin in the dynamics of quenching.

To identify factors governing peel-color development in mandarins, we examined carotenoid content and composition and the expression of carotenoid-related genes during four stages of ripening (i.e., green, breaker, yellow, and orange) in two varieties: 'Ora', which has orange fruit, and 'Shani', which has orange-reddish fruit. The two varieties had different carotenoid compositions, and 'Shani' had a significantly higher level of total carotenoid pigments. 'Shani' was rich in the deep orange β-cryptoxanthin and the orange-reddish β-citraurin, whereas 'Ora' was rich in the orange violaxanthin. RNA-Seq analysis revealed significantly greater expression of the carotenoid-biosynthesis genes PSY, βLCY, βCHX, and CCD4b, as well as MEP-pathway genes and several ethylene-biosynthesis and -signaling genes in 'Shani' fruit. In contrast, the expression levels of genes involved in the synthesis of α-branch carotenoids (i.e., εLCY and εCHX) and ZEP, which is involved in the formation of violaxanthin, were significantly higher in the 'Ora' fruit.

We tested the potential of Raman microspectroscopy to determine carotenoid pigments — both primary (lutein, beta-carotene) and secondary (astaxanthin) carotenoids — in the different species and life-cycle stages of snow algae from the order Chlamydomonadales (Chlorophyta). We compared the performance of Raman spectrometry to a reference method of biological pigment analysis, high-performance liquid chromatography (HPLC). The three main carotenoid Raman bands of the astaxanthin-rich red cysts were located at 1520, 1156 and 1006 cm−1. The shifts (orange aplanozygotes and green motile cells with flagella) in the position of the ν1(C C) Raman band of the polyenic chain is consistent with the expected changes in the ratios of the various carotenoid pigments. Flagellated green cells commonly contain lutein as a major carotenoid, together with minor amounts of β‑carotene and varying amounts of antheraxanthin, violaxanthin and neoxanthin. Aplanozygotes contain mixtures of both primary and secondary carotenoids. In most cases, the ν1(C C) band is an overlapping set of bands, which is due to the signal of all carotenoid pigments in the sample, and a deconvolution along with the band position shifts (mainly ν1) could be used to characterize the mixture of carotenoids. However, the ability of Raman spectroscopy to discriminate between structurally slightly differing carotenoid pigments or several carotenoids in an admixture in an unknown biological system remains limited.

Los árboles del género Paulownia son de rápido crecimiento y tienen importancia económica en Asia como materia prima de uso común en la elaboración de muebles, instrumentos musicales y cercos, lo que ha despertado interés por cultivarlos en diferentes ambientes. Con el objetivo de evaluar la tolerancia al estrés hídrico se eligieron tres especies, P. imperialis, P. fortunei y P. elongata, porque son las que se utilizan en China con mayor frecuencia en la reforestación y en la industria maderera. Se realizaron análisis bioquímicos de las hojas para determinar los siguientes componentes: como pigmentos, a las clorofilas total, a y b; β-caroteno, violaxantina y luteína); las enzimas peroxidasa y prolina, así como proteínas solubles, insolubles y totales en tres diferentes condiciones de humedad del suelo, bajo dos condiciones ambientales, campo e invernadero, entre los cuales se observaron diferencias significativas; destaca un incremento del contenido de pigmentos y peroxidasa, y una disminución de las proteínas y prolina en el ambiente controlado, principalmente. Al evaluar la respuesta al estrés hídrico entre las especies se concluyó que P. imperialis y P. elongata son más tolerantes que P. fortunei, debido a su mayor contenido de prolina, mas proteínas totales y solubles, indicadores de una mejor tolerancia a condiciones de estrés.

The microalgal strain, JNU5, was isolated downriver from the Hengshishui River of the Guangdong Province, China. The morphology of JNU5 revealed that it was a spherical, unicellular organism with a smooth cell wall. The cell contained a large vacuole and prominent, reddish globule. The pigment composition revealed that chlorophyll a was the only chlorophyll, and violaxanthin was the predominant carotenoid along with vaucheriaxanthin-ester and β-carotene. The fatty acid profile of JNU5 mainly consisted of myristic acid, palmic acid, linoleic acid, and eicosapentaenoic acid (EPA). Moreover, the EPA content consisted of up to 40% of total fatty acids. Phylogeny was determined using the 18S rDNA sequences suggested that JNU5 was a member of Eustigmatophyceae, belonging to Clade IIa and was close to Trachydiscus minutus. T. minutus and JNU5 were easily distinguished by cell shape and size. This indicates that JNU5 is a new species, and thus, we have named it, Trachydiscus guangdongensis sp. nov.. Meanwhile, the cell wall composition of JNU5 was analyzed and was found to be mainly composed of galactose (66%) with some glucose and mannose present. The water-soluble polysaccharide (WSP) of the cell wall was primarily composed of galactose, which accounted for 90% of total sugar.

Purpose The purpose of this study is to evaluate the pattern of carotenoids distribution in three underutilized Malaysian ‘ulam’ or traditional vegetables in Malaysia (Averrhoa carambola, Manihot esculenta and Ipomoea batatas) and their valuable pro-vitamin A activities. These assessments will yield valuable knowledge and insight into the importance of these underutilized traditional vegetables and highlight their potential for applications in medicinal and functional colorant industries. Design/methodology/approach In this study, the authors have evaluated the distribution of carotenoid compounds in aerial organs of three underutilized traditional vegetables (Averrhoa carambola, Manihot esculenta and Ipomoea batatas). The content of the individual carotenoids were quantified using high-performance liquid chromatography (HPLC) and the organ with the highest accumulation of these compounds were identified. Their valuable pro-vitamin A activities were also reported to indicate their medicinal potential which can further be exploited as pharmacologically active natural colorants and in other applications. Findings In total, three major chromatographic peaks corresponding to lutein, violaxanthin and β-carotene were observed through HPLC. Among the compounds detected, lutein and β-carotene were the most abundant carotenoids found in both shoots and petioles of all three species. Violaxanthin was only detected in I. batatas shoots. Overall, carotenoid content was observed to be higher in the shoots than in the petioles, where I. batatas contained the highest amount of total carotenoid, followed by M. esculenta and A. carambola. The opposite trend was observed in the petioles, where A. carambola petioles had the highest carotenoid content, while I. batatas contained the least. Research limitations/implications The distribution and abundance of these individual carotenoids suggested that the petioles contained the highest amount of carotenoid, contributing to its high pro-vitamin A activity, and could be potentially useful for medicinal application, as it can act as storage site that is not as prone to natural drying or degradation during harvest and sample storage. Future research work should include improvements in the extraction and purification procedures as well as robust identification methods which may lead to better detection and identification of other compounds that could attribute to its bioactivity, to complement the findings of the current study. Practical implications This analysis provides valuable information on the importance of underutilized traditional vegetables as important biofactories for sustainable production of valuable pigments (such as carotenoids) with medicinal benefits and can further be exploited in various industries, such as in formulation of functional natural colorants. This study also highlights the importance of petiole as a storage site of pharmacologically active compounds that is not as prone to natural drying or degradation during harvest and sample storage. Originality/value To date, there is no previous report found on comparative analysis of carotenoid content and quantification of individual carotenoid concentration in the edible aerial parts of Averrhoa carambola, Manihot esculenta and Ipomoea batatas, although they have been traditionally consumed as “ulam” in Malaysia. Therefore, the results reported in this study provide new insights on carotenoid accumulation in the selected ‘ulam’ species.

We studied the effects of different radiation treatments on the physiology and UV-absorbing compounds of the model liverwort Marchantia polymorpha subsp. ruderalis. Starting from gemmae, samples were exposed to five radiation treatments: low photosynthetically active radiation (PAR), low PAR+ UV-A, low PAR + UV-B, low PAR + UV-A + UV-B, and high PAR. After 35 days, the maximum quantum yield of photosystem II was similar between treatments, which suggested comparable photoinhibition and physiological vitality, also supported by results showing an unchanged chlorophyll a/b ratio and only slight changes in growth. However, the total contents of both chlorophylls and carotenoids decreased in the UV radiation treatments and, more strongly, in the high-PAR samples, suggesting mainly PAR-dependent damage to the photosynthetic pigments. The xanthophyll index (antheraxanthin + zeaxanthin)/(violaxanthin + antheraxanthin + zeaxanthin) was only increased in the high-PAR samples, indicating an increase in photoprotection through nonphotochemical dissipation of the excess energy. The sclerophylly index (the ratio between the thallus dry mass and surface area) was increased in the UV-B-exposed samples, suggesting a UV-induced structural protection. Only the UV-B-exposed samples showed DNA damage. Several apigenin and luteolin derivatives were found in the methanol-soluble vacuolar fraction of the liverwort and p-coumaric and ferulic acids in the methanol-insoluble cell wall-bound fraction. Most individual soluble compounds, the bulk level of soluble compounds, and chalcone synthase expression increased in UV-B-exposed samples, whereas individual insoluble compounds increased in the samples exposed to only PAR. Principal components analysis summarized these responses, showing the strong influence of both UV-B and PAR levels on the physiology and UV protection of the samples.

Photosynthetic eukaryotes whose cells harbor plastids originating from secondary endosymbiosis of a red alga include species of major ecological and economic importance. Since utilization of solar energy relies on the efficient light-harvesting, one of the critical factors for the success of the red lineage in a range of environments is to be found in the adaptability of the light-harvesting machinery, formed by the proteins of the light-harvesting complex (LHC) family. A number of species are known to employ mainly a unique class of LHC containing red-shifted chlorophyll a (Chl a) forms absorbing above 690 nm. This appears to be an adaptation to shaded habitats. Here we present a detailed investigation of excitation energy flow in the red-shifted light-harvesting antenna of eustigmatophyte Trachydiscus minutus using time-resolved fluorescence and ultrafast transient absorption measurements. The main carotenoid in the complex is violaxanthin, hence this LHC is labeled the red-violaxanthin-Chl a protein, rVCP. Both the carotenoid-to-Chl a energy transfer and excitation dynamics within the Chl a manifold were studied and compared to the related antenna complex, VCP, that lacks the red-Chl a. Two spectrally defined carotenoid pools were identified in the red antenna, contributing to energy transfer to Chl a, mostly via S2 and hot S1 states. Also, Chl a triplet quenching by carotenoids is documented. Two separate pools of red-shifted Chl a were resolved, one is likely formed by excitonically coupled Chl a molecules. The structural implications of these observations are discussed.