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The physicochemical and antioxidant properties of seven carotenoids: antheraxanthin, β-carotene, neoxanthin, peridinin, violaxanthin, xanthrophyll and zeaxanthin were studied by theoretical means. Then the Optoelectronic properties and interaction of chlorophyll-carotenoid complexes are analysed by TDDFT and IGMPLOT. Global reactivity descriptors for carotenoids and chlorophyll (Chla, Chlb) are calculated via conceptual density functional theory (CDFT). The higher HOMO–LUMO (HL) gap indicated structural stability of carotenoid, chlorophyll and chlorophyll-carotenoid complexes. The chemical hardness for carotenoids and Chlorophyll is found to be lower in the solvent medium than in the gas phase. Results showed that carotenoids can be used as good reactive nucleophile due to lower µ and ω. As proton affinities (PAs) are much lower than the bond dissociation enthalpies (BDEs), it is anticipated that direct antioxidant activity in these carotenoids is mainly due to the sequential proton loss electron transfer (SPLET) mechanism with dominant solvent effects. Also lower PAs of carotenoid suggest that antioxidant activity by the SPLET mechanism should be a result of a balance between proclivities to transfer protons. Reaction rate constant with Transition-State Theory (TST) were estimated for carotenoid-Chlorophyll complexes in gas phase. Time dependent Density Functional Theory (TDDFT) showed that all the chlorophyll (Chla, Chlb)–carotenoid complexes show absorption wavelength in the visible region. The lower S1–T1 adiabatic energy gap indicated ISC transition from S1 to T1 state.

Allelopathic activities of three carotenoids of a natural pigment group, neoxanthin, crocin and β-carotene, were assayed by the protoplast co-culture method with digital image analysis (DIA-PP method). Effects on three different growth stages of lettuce protoplasts, i.e., cell wall formation, cell division, and yellow pigment accumulation, were investigated using 96-well culture plates. Cell division was inhibited 65–95% by all three carotenoids at 33–100 µM. Inhibition of cell division stage was stronger than at the cell wall formation stage in neoxanthin, and the water-soluble carotenoid, crocin, whose yellow pigment was incorporated into the vacuole of lettuce protoplasts. Neoxanthin at 33 µM and crocin at higher than 100 µM inhibited more than 100% of the yellow pigment accumulation. By contrast, at low concentrations (0.01–1 µM) β-carotene stimulated growth at the cell division stage. At high concentrations of β-carotene (100–500 µM), inhibition was prominent at all three stages, and also in neighboring wells of zero control, which suggested emission of a volatile compound by β-carotene. They were compared with the report of the volatile compound, tulipalin A. Differences in patterns of inhibition of carotenoids on lettuce protoplast growth were compared with reports of another natural pigment, anthocyanin, and anthocyanin-containing red callus cultured in the light, and with that of neoxanthin-containing yellow callus cultured in the dark.

Potato tubers are an important agricultural crop containing carotenoids, an important group of secondary plant metabolites showing positive health benefits. This paper examines nine carotenoids at weekly intervals in three cultivars with different flesh color during the tuber maturation process, as analyzed by HPLC-DAD. Total carotenoids showed a bell-shaped distribution with maximum values in the middle of the tuber maturation process rather than in ready-to-harvest tubers. Xanthophylls (viola-, anthera-, neoxanthin and lutein) were the predominant carotenoid forms found in the cultivars. Red and purple-fleshed cultivars contained only a quarter of the total carotenoids of a commercial cultivar with yellow flesh. Weather conditions moderately influenced the carotenoids of the xanthophyll cycle. In red or purple-fleshed tubers no relationship was found between total carotenoids and total anthocyanidins.

Marigold (Tagetes erecta L.) is an important ornamental plant with a wide variety of flower colors. Despite its economic value, few biochemical and molecular studies have explored the generation of flower color in this species. To study the mechanism underlying marigold petal color, we performed a metabolite analysis and de novo cDNA sequencing on the inbred line ‘V-01’ and its petal color mutant ‘V-01M’ at four flower developmental stages. A total of 49,217 unigenes were identified from 24 cDNA libraries. Based on our metabolites and transcriptomic analyses, we present an overview of carotenoid biosynthesis, degradation, and accumulation in marigold flowers. The carotenoid content of the yellow mutant ‘V-01M’ was higher than that of the orange inbred line ‘V-01’, and the abundances of the yellow compounds lutein, neoxanthin, violaxanthin, zeaxanthin, and antheraxanthin were significantly higher in the mutant. During flower development, the carotenoid biosynthesis genes were upregulated in both ‘V-01’ and ‘V-01M’, with no significant differences between the two lines. By contrast, the carotenoid degradation genes were dramatically downregulated in the yellow mutant ‘V-01M’. We therefore speculate that the carotenoid degradation genes are the key factors regulating the carotenoid content of marigold flowers. Our research provides a large amount of transcriptomic data and insights into the marigold color metabolome.

Leaves are primary plant organs harbouring chloroplasts that enable the sensing of changes in environmental conditions and contribute to maintaining cellular homeostasis. Carotenoids, carotenoid-derived phytohormones and apocarotenoid metabolites facilitate photosynthesis, photoprotection and stress acclimation in leaves. The thylakoids in the chloroplasts from the photosynthetically functional leaves accumulate a specific composition of lutein, β-carotene, violaxanthin, and neoxanthin that act as accessory pigments to chlorophyll and maintain optimal photosynthetic performance. The developmental state of leaves and environmental conditions can affect carotenoid metabolism and, hence, chloroplast functions in leaves. The etioplasts in the cotyledons from the germinating seedlings abundantly accumulate lutein. The light exposure of cotyledons triggers ultrastructural development of chloroplast accumulating all primary foliar carotenoids and chlorophylls. The accumulation of zeaxanthin and antheraxanthin during intense light and high temperature maintains the functional integrity of thylakoid membranes and chloroplasts. Carotenoids are continuously synthesised and degraded in leaves, yet their composition remains tightly maintained. The disintegration of chloroplast, such as during the leaf senescence, ceases the carotenoid biosynthesis and triggers carotenoid degradation. This review sheds light on the fundamental aspects of carotenoid metabolism. The effects of altered light regimes, carbon dioxide (CO2), temperature, drought, and soil nutrient levels are discussed in the context of carotenoid metabolism in leaves. The outstanding questions on how environmental perturbations can alter carotenoid metabolism and chloroplast development in leaves are highlighted. This review provides further insights on how carotenoid regulatory networks could be engineered to enrich carotenoids in the leaves of food and horticultural crops.

In this study, the microalga Chlorococum humicula was evaluated as a potential source of carotenoids and chlorophylls, and extraction of these pigments with liquefied dimethyl ether (DME) as an alternative green solvent was investigated. Carotenoids and chlorophylls in the algae were identified by spectral and comparative band ratio analyses. The liquefied DME extraction was shown to be a rather fast process for the extraction of pigments from C. humicula at relatively high yields. The effects of the extraction solvent: sample ratio, time, and temperature in the liquefied DME extraction of C. humicola on the yields of neoxanthin, violaxanthin, lutein, chlorophyll-b, chlorophyll-a, as well as the total carotenoids and chlorophylls, were determined. The highest total carotenoids yield (4.14 mg/g dry weight algae) and total chlorophylls yield (8.45 mg/g dry weight algae) were obtained using a 45:1 (w/w) liquefied DME: algae wet weight ratio for 20 min at 41 °C. At this condition, extraction of 91 % and 35 % of the total carotenoids and chlorophylls, respectively, was attained. In contrast, the extraction yield and percentage recovery of carotenoids obtained with acetone, a conventional solvent, was found to be lower than that of the liquefied DME, suggesting that liquefied DME has a higher selectivity for the less polar carotenoids than that of acetone.

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).

In this study, two carotenoid antioxidants lutein and neoxanthin were investigated for their ability to prevent oxidatively induced DNA damage. A Chlamydomonas reinhardtii alga was used as the natural source of lutein and neoxanthin. Analysis using HPLC with Diode Array Detector (DAD) showed that lutein and neoxanthin were the most abundant carotenoids in the extract of C. reinhardtii. An analysis using GC–MS/MS was performed to measure the damage induced by Fenton reaction to DNA and to evaluate the ability of lutein, neoxanthin and C. reinhardtii extract to inhibit such damage. The results showed that lutein and neoxanthin have the ability to reduce oxidatively induced DNA base damage to less than half of its magnitude. Extract of C. reinhardtii inhibited about 76% of the damage in comparison to control sample. Neoxanthin in lower concentrations than those of lutein proved to be better inhibitor of oxidatively induced DNA damage.

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.

Light is the most essential environmental factor that plants require for the growth and development. However, light is highly heterogeneous in natural condition, and plants have to evolve a series of strategies to acclimate the dynamic light since it is sessile. The photosynthetic performance and regulatory mechanisms involved in acclimation were elucidated in shade-tolerant plant Amorphophallus xiei under steady state and fluctuating high light. The dissipation and allocation of light energy, photosynthetic capacity, and antioxidant defense were examined in A. xiei cultivated under 4%, 17%, and 100% of full sunlight. High-light-grown plants exhibited reduced photosynthesis and the slowest response to simulated sunflecks than the other two treatments. Maximum and actual efficiency of PSII photochemistry (Fv′/Fm′ and ΦPSII) and electron transport rate (ETR) were lowest, and non-photochemical quenching (NPQ) were highest in high-light-grown plants than in low- and intermediate-light-grown plants subjected to different photon flux densities (PFD) and intercellular CO2 concentrations (Ci). Fv′/Fm′, ΦPSII, ETR and NPQ were lowest in high-light-grown A. xiei than others, when plants were exposed to simulated sunflecks. In fully light-induced leaves, high-light-grown plants showed a maximum value in quantum efficiency of light-dependent thermal dissipation (ΦNPQ), and a minimum value in ΦPSII was recorded. Low-light-grown plants subjected to simulated sunflecks, a maximum value in ΦNPQ and ΦPSII were observed. Mass-based nitrogen content (Narea), specific leaf area (SLA), superoxide dismutase (SOD) and catalase (CAT) activities significantly declined with the increase of growth irradiance. The high-light-grown plants showed higher xanthophyll de-epoxidation and ascorbate–glutathione cycle activity, while low-light-grown plants showed higher neoxanthin and β-carotene contents. In general, high-light-grown A. xiei generally show a depressed photosynthetic capacity, and are efficient in dissipating excess light energy by NPQ under a steady-state light but not under a highly fluctuating light. Low-light-grown plants show a rapid photosynthetic assimilation response to sunflecks and then a rapid activation of the energy-dependent quenching (qE) component of NPQ under the sunflecks. In addition, high-light-grown plants, although using various strategies to reduce light absorption and to scavenge reactive oxygen species, have less efficient protection against photodamage.

Chinese kale (Brassica oleracea var. alboglabra) is rich in carotenoids, and neoxanthin is one of the most important carotenoids in Chinese kale. In this study, the function of the neoxanthin synthase gene (BoaNXS) in Chinese kale was investigated. BoaNXS, which had a 699-bp coding sequence, was cloned from the white flower cultivar of Chinese kale and was expressed in all developmental stages and organs of Chinese kale, its expression was highest in young seeds. The subcellular localization indicated that BoaNXS was localized in the chloroplast. BoaNXS-overexpressed plants were obtained via Agrobacterium-mediated transient overexpression methodology, and the gene overexpression efficiencies ranged from 2.10- to 4.24-fold. The color in the leaves of BoaNXS-overexpressed plants changed from green to yellow-green, the content of total and individual carotenoids, such as neoxanthin, violaxanthin, and lutein, was significantly increased, and the expression levels of most carotenoid biosynthetic genes were notably increased. These findings indicated that BoaNXS is of vital importance in carotenoid biosynthesis in Chinese kale and could be used as a candidate gene for enriching the carotenoid accumulation and color of Chinese kale and other Brassica vegetables.

The dietary needs of humans for provitamin A carotenoids arise from their inability to synthesize vitamin A de novo. To improve the status of this essential micronutrient, special attention has been given to biofortification of staple foods, such as wheat grains, which are consumed in large quantities but contain low levels of provitamin A carotenoids. However, there remains an unclear contribution of metabolic genes and homoeologs to the turnover of carotenoids in wheat grains. To better understand carotenoid catabolism in tetraploid wheat, Targeting Induced Local Lesions in Genomes (TILLING) mutants of CCD4, encoding a Carotenoid Cleavage Dioxygenase (CCD) that cleaves carotenoids into smaller apocarotenoid molecules, were isolated and characterized. Our analysis showed that ccd4 mutations co-segregated with Poltergeist-like (pll) mutations in the TILLING mutants of A and B subgenomes, hence the ccd-A4 pll-A, ccd-B4 pll-B, and ccd-A4 ccd-B4 pll-A pll-B mutants were analyzed in this study. Carotenoid profiles are comparable in mature grains of the mutant and control plants, indicating that CCD4 homoeologs do not have a major impact on carotenoid accumulation in grains. However, the neoxanthin content was increased in leaves of ccd-A4 ccd-B4 pll-A pll-B relative to the control. In addition, four unidentified carotenoids showed a unique presence in leaves of ccd-A4 ccd-B4 pll-A pll-B plants. These results suggested that CCD4 homoeologs may contribute to the turnover of neoxanthin and the unidentified carotenoids in leaves. Interestingly, abnormal spike, grain, and seminal root phenotypes were also observed for ccd-A4 pll-A, ccd-B4 pll-B, and ccd-A4 ccd-B4 pll-A pll-B plants, suggesting that CCD4 and/or PLL homoeologs could function toward these traits. Overall, this study not only reveals the role of CCD4 in cleavage of carotenoids in leaves and grains, but also uncovers several critical growth traits that are controlled by CCD4, PLL, or the CCD4-PLL interaction.

Persistent pollutants such as pharmaceuticals, pesticides, musk fragrances, and dyes are frequently detected in different environmental compartments and negatively impact the environment and humans. Understanding the impacts of diffuse environmental pollutants on plants is still limited, especially at realistic environmental concentrations of contaminants. We studied the effects of key representatives of two major classes of environmental pollutants (nine different antibiotics and six different textile dyes) on the leaf carotenoid (violaxanthin and neoxanthin) content in wheat (Triticum aestivum L.) using different pollutant concentrations and application times. The wheat plants were watered with solutions of selected environmental pollutants in two different concentrations of 0.5 mg L−1 and 1.5 mg L−1 for one week (0.5 L) and two weeks (1 L). Both categories of pollutants selected for this study negatively influenced the content of violaxanthin and neoxanthin, whereas the textile dyes represented more severe stress to the wheat plants. The results demonstrate that chronic exposure to common diffusively spread environmental contaminants constitutes significant stress to the plants.

To reveal the molecular mechanism underlying peel colouration, carotenoid metabolites and the transcriptome were jointly analysed in zucchini peels with three different colours: light green (Lg), yellow (Y), and orange (O). Our results showed that the carotenoid levels in O (157.075 μg/g) and Y (22.734 μg/g) were both significantly higher than in Lg (7.435 μg/g), while the chlorophyll content was highest in Lg (32.326 μg/g), followed by O (7.294 μg/g) and Y (4.617 μg/g). A total of 14 carotenoids were detected in zucchini peels, primarily lutein (103.167 μg/g in Lg, 509.667 μg/g in Y, and 1543.333 μg/g in O). In particular, significant accumulation of antheraxanthin, zeaxanthin, neoxanthin, and β-cryptoxanthin was first reported in orange zucchini in this study. Furthermore, two modules with hub genes related to carotenoid or chlorophyll content were identified through weighted gene coexpression network analysis. Additionally, the transcription level of some hub genes (PIF4, APRR2, bHLH128, ERF4, PSY1, LCYE2, and RCCR3) was highly correlated with pigment content in the peel, which may be responsible for carotenoid accumulation and chlorophyll degradation in the Y and O varieties. Taken together, the results obtained in this study help to provide a novel mechanism underlying peel colouration in zucchini.

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.

Seasonal accumulation of hormonal and photoprotective isoprenoids, particularly α-tocopherol, carotenoids and abscisic acid, indicate their important role in protecting Cistus albidus plants from environmental stress during a Mediterranean winter. The high diurnal amounts of α-tocopherol and xanthophylls 3 h before maximum light intensity suggest a photoprotective response against the prevailing diurnal changes. The timing to modulate acclimatory/defense responses under changing environmental conditions is one of the most critical points for plant fitness and stress tolerance. Here, we report seasonal and diurnal changes in the contents of isoprenoids originated from the methylerythritol phosphate pathway, including chlorophylls, carotenoids, tocochromanols, and phytohormones (abscisic acid, cytokinins, and gibberellins) in C. albidus during a Mediterranean winter. Plants were subjected not only to typically low winter temperatures but also to drought, as shown by a mean plant water status of 54% during the experimental period. The maximum PSII efficiency, however, remained consistently high (Fv/Fm > 0.8), proving that C. albidus had efficient mechanisms to tolerate combined stress conditions during winter. While seasonal α-tocopherol contents remained high (200–300 µg/g DW) during the experimental period, carotenoid contents increased during winter attaining maximum levels in February (minimum air temperature ≤ 5 °C for 13 days). Following the initial transient increases of bioactive trans-zeatin (about fivefold) during winter, the increased abscisic acid contents proved its important role during abiotic stress tolerance. Diurnal amounts of α-tocopherol and xanthophylls, particularly lutein, zeaxanthin and neoxanthin including the de-epoxidation state, reached maximum levels as early as 2 h after dawn, when solar intensity was 68% lower than the maximum solar radiation at noon. It is concluded that (1) given their proven antioxidant properties, both α-tocopherol and carotenoids seem to play a crucial role protecting the photosynthetic apparatus under severe stress conditions; (2) high seasonal amounts of abscisic acid indicate its important role in abiotic stress tolerance within plant hormones, although under specific environmental conditions, accumulation of bioactive cytokinins appears to be involved to enhance stress tolerance; (3) the concerted diurnal adjustment of α-tocopherol and xanthophylls as early as 3 h before maximum light intensity suggests that plants anticipated the predictable diurnal changes in the environment to protect the photosynthetic apparatus.

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.

Fourteen watermelon cultivars with different fruit flesh colors (red, salmon yellow, orange, and canary yellow) were analyzed for carotenoid contents (prolycopene, lycopene, β-carotene, ζ-carotene, and neoxanthin). Genes encoding the carotenoid biosynthesis enzymes carotenoid isomerase (encoded by CRTISO), which catalyzes the isomerization of prolycopene to lycopene, and β-carotene hydroxylase (CHYB), which catalyzes the conversion of β-carotene to xanthophyll, were also analyzed. High-performance liquid chromatography showed that the salmon yellow and orange flesh accumulated either prolycopene (orange-P flesh) or β-carotene (orange-β flesh), whereas lycopene and neoxanthin were the main carotenoids in the red and canary yellow flesh, respectively. Quantitative real-time polymerase chain reaction indicated that CRTISO and CHYB were mainly expressed during fruit maturation, regardless of the flesh color, and there was no significant association between differential gene expression and flesh color. Importantly, transcript sequencing revealed a non-synonymous single-nucleotide mismatch (T > C1976) in exon 13 of CRTISO between orange-P-fleshed and other cultivars, suggesting CRTISO as a candidate gene for high prolycopene accumulation. However, in β-carotene-accumulating cultivars, there were no mutations in CHYB transcripts. A cleaved amplified polymorphic sequence marker was developed for T > C1976, and its applicability for marker-assisted selection of orange-P flesh was validated in 105 watermelon accessions.

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.

Some carotenoids are found in the Euglena gracilis, including β-carotene, diadinoxanthin, diatoxanthins, and neoxanthin as the major species; however, the molecular mechanism underlying carotenoid biosynthesis in E. gracilis is not well understood. To clarify the pathway and regulation of carotenoid biosynthesis in this alga, we functionally characterized the cytochrome P450 (CYP)-type carotene hydroxylase gene EgCYP97H1. Heterologous in vivo enzyme assay in E. coli indicated that EgCYP97H1 hydroxylated β-carotene to β-cryptoxanthin. E. gracilis cells suppressing EgCYP97H1 resulted in marked growth inhibition and reductions in total carotenoid and chlorophyll contents. Analysis of carotenoid composition revealed that suppression of EgCYP97H1 resulted in higher level of β-carotene, suggesting that EgCYP97H1 is physiologically essential for carotenoid biosynthesis and thus normal cell growth. To our knowledge, this is the first time EgCYP97H1 has been suggested to be β-carotene monohydroxylase, but not β-carotene dihydroxylase. Moreover, during light adaptation of dark-grown E. gracilis, transcript levels of the carotenoid biosynthetic genes (EgCYP97H1, geranylgeranyl pyrophosphate synthase EgcrtE, and phytoene synthase EgcrtB) remained virtually unchanged. In contrast, carotenoid accumulation in E. gracilis grown under the same conditions was inhibited by treatment with a translational inhibitor but not a transcriptional inhibitor, indicating that photo-responsive carotenoid biosynthesis is regulated post-transcriptionally in this alga.

The foremost problem in carotenoid research is the excessive cost and difficulty of maintaining pure carotenoid compounds. This work presents an economical, efficient, and simplified one-step scheme for the purification of four major xanthophyll carotenoids from lettuce by utilizing preparative thin layer chromatography on Hyflo-Super-Cel: MgO (Heavy): calcium sulfate hemihydrate (9:9:2 w/w) based adsorbent. The mobile phase of acetone: hexane (1:1) provided the perfect separation of major xanthophylls, resulting in 95-96% purity after just single-step separation, with no interference from chlorophylls or other minor carotenoids. The identity of carotenoids was confirmed by absorption spectroscopy, chemical tests and APCI+-MS/MS. The proposed scheme can be used to isolate the carotenoids at the analytical and preparative scale. In anticancer studies, among four xanthophylls, 9-Z-neoxanthin was found most potent for reduction of cell viability of cervical (HeLa) and lung cancer (A549) cells, with IC50 values of 3.8 and 7.5 μM, respectively.

The present study aimed to investigate the effect of young leaves on fat accumulation in differentiated 3T3-L1 adipocytes. A potent preventive effect on fat accumulation was observed in fractions of young leaves of spinach, beet, and arugula extracted with a low-polarity solvent (hexane:acetone:ethanol:toluene = 10:6:7:6). This effect was seemingly associated with the leaf carotenoid content, including lutein, β-carotene, and neoxanthin. Among these, only neoxanthin, with the characteristic structure of 5,6-monoepoxide and an allenic bond, significantly prevented fat accumulation in a dose-dependent manner. The preventive effect and carotenoid content, including neoxanthin, of these young leaves did not differ from those of the corresponding adult leaves. Therefore, our study demonstrated that young vegetable leaves, such as spinach, beet, and arugula leaves, contained neoxanthin, which prevented fat accumulation in adipocytes in vitro. In the future, the effectiveness of such young leaves and neoxanthin should be investigated in vivo.

Summary • Here, the effects of salinity (0–400 mM NaCl) on photosynthesis, photosystem II (PSII) efficiency and the xanthophyll cycle were investigated in the halophyte Atriplex centralasiatica grown under outdoor conditions. • Leaf sodium and chloride in leaves increased considerably whereas CO2 assimilation rate decreased. PSII efficiency (ΦPSII) and the efficiency of excitation energy capture by open PSII reaction centres decreased whereas nonphotochemical quenching (NPQ) increased significantly. There was no change in photochemical quenching (qP) in salt-adapted plants. Salinity induced no changes in the maximum efficiency of PSII photochemistry (Fv/Fm) measured either at midday or at predawn. However, Fv/Fm values were c. 20% lower at midday than at predawn. • Contents of chlorophyll (a + b), neoxanthin, lutein and β-carotene were unchanged with increasing salt concentration, but zeaxanthan increased significantly, at the expense of violaxanthin. There was a linear relationship between the de-epoxidation state of the xanthophyll cycle and ΦPSII, , and NPQ. • Our results suggest that A. centralasiatica shows high tolerance to both high salinity and photoinhibition and that the xanthophyll cycle played an important role in protecting photosynthetic apparatus from photoinhibitory damage. Tolerance of PSII to salinity and photoinhibition can be viewed as an important strategy for A. centralasiatica to grow in very high saline soil during the summer season with high irradiance.

In this study, various ratios of combined red, blue, and amber light-emitting diodes (rbaLEDs) were investigated for their effect on the expression of carotenoid biosynthetic genes and carotenoid accumulation in eight Brassica microgreens. Total and individual (β-carotene, lutein, α-carotene, neoxanthin, and violaxanthin) carotenoids were increased 20-44 and 10-55%, respectively, under dose-dependent increasing amber-blue light and decreasing red in most microgreens. Lipophilic 2,2-diphenyl-1-picrylhydrazyl and ferric reducing antioxidant power antioxidant activities were significantly increased under higher amber and blue light fractions, while oxygen radical absorbance capacity was generally decreased. Under rbaLED in mizuna (B. rapa) microgreens, the lycopene epsilon cyclase (LYCe) expression was 10-15-fold higher, which resulted in downstream accumulation of α-carotene and lutein. Lycopene beta cyclase (LYCβ) was not significantly changed, suggesting that β-carotene, violaxanthin and neoxanthin were mainly controlled by upstream phytoene synthase and branch-point LYCe. Increased beta-ring carotenoid hydroxylase (CHXβ) expression was also consistent with lutein accumulation. This study demonstrated for the first time that amber LED was involved in the regulatory mechanism of carotenoid biosynthesis, thus a potential novel approach to production of antioxidant-rich microgreens.

Key message: The suppression of the HYD-1 gene by a TILLING approach increases the amount of ?-carotene in durum wheat kernel. Abstract: Vitamin A deficiency is a major public health problem that affects numerous countries in the world. As humans are not able to synthesize vitamin A, it must be daily assimilated along with other micro- and macronutrients through the diet. Durum wheat is an important crop for Mediterranean countries and provides a discrete amount of nutrients, such as carbohydrates and proteins, but it is deficient in some essential micronutrients, including provitamin A. In the present work, a targeting induced local lesions in genomes strategy has been undertaken to obtain durum wheat genotypes biofortified in provitamin A. In detail, we focused on the suppression of the ?-carotene hydroxylase 1 (HYD1) genes, encoding enzymes involved in the redirection of ?-carotene toward the synthesis of the downstream xanthophylls (neoxanthin, violaxanthin and zeaxanthin). Expression analysis of genes involved in carotenoid biosynthesis revealed a reduction of the abundance of HYD1 transcripts greater than 50% in mutant grain compared to the control. The biochemical profiling of carotenoid in the wheat mutant genotypes highlighted a significant increase of more than 70% of ?-carotene compared to the wild-type sibling lines, with no change in lutein, ?-carotene and zeaxanthin content. This study sheds new light on the molecular mechanism governing carotenoid biosynthesis in durum wheat and provides new genotypes that represent a good genetic resource for future breeding programs focused on the provitamin A biofortification through non-transgenic approaches.

This study aimed to evaluate the effect of dynamic red and blue light parameters on the physiological responses and key metabolites in lettuce and also the subsequent impact of varying light spectra on nutritive value. We explored the metabolic changes in carotenes, xanthophylls, soluble sugars, organic acids, and antioxidants; the response of photosynthetic indices [photosynthetic (Pr) and transpiration (Tr) rates]; and the intracellular to ambient CO2 concentration ratios (Ci/Ca) in lettuce (Lactuca sativa L. “Lobjoits Green Cos”). They were cultivated under constant (con) or parabolic (dyn) blue (B, 452 nm) and/or red (R, 662 nm) light-emitting diode (LED) photosynthetic photon flux densities (PPFDs) at 12, 16, and 20 h photoperiods, maintaining consistent daily light integrals (DLIs) for each light component in all treatments, at 2.3 and 9.2 mol m–2 per day for blue and red light, respectively. The obtained results and principal component analysis (PCA) confirmed a significant impact of the light spectrum, photoperiod, and parabolic profiles of PPFD on the physiological response of lettuce. The 16 h photoperiod resulted in significantly higher content of xanthophylls (neoxanthin, violaxanthin, lutein, and zeaxanthin) in lettuce leaves under both constant and parabolic blue light treatments (BconRdyn 16 h and BdynRdyn 16 h, respectively). Lower PPFD levels under a 20 h photoperiod (BdynRdyn 20 h) as well as higher PPFD levels under a 12 h photoperiod (BdynRdyn 12 h) had a pronounced impact on leaf gas exchange indices (Pr, Tr, Ci/Ca), xanthophylls, soluble sugar contents, and antioxidant properties of lettuce leaves. The parabolic PPFD lighting profile over a 16 h photoperiod (BdynRdyn 16 h) led to a significant decrease in Ci/Ca, which resulted in decreased Pr and Tr, compared with constant blue or red light treatments with the same photoperiod (BconRdyn and BdynRcon 16 h). Additionally, constant blue lighting produced higher α + β-carotene and anthocyanin (ARI) content and increased carotenoid to chlorophyll ratio (CRI) but decreased biomass accumulation and antioxidant activity.

The effect of steam and sous-vide oven procedures on liposoluble antioxidants of colored cauliflower (orange and purple) was assessed for the first time and compared with domestic practice (boiling). In raw samples, the total carotenoid content was 10-fold higher in Cheddar than in Depurple (20.9 ± 2.1 vs. 2.3 ± 0.5 mg/kg dry weight), whereas the level of tocopherols was similar (28.5 ± 4.4 vs. 33 ± 5.2 mg/kg dry weight). The Cheddar liposoluble antioxidant matter contained violaxanthin, neoxanthin, α-carotene and δ-tocopherol, not detected in Depurple. All tests increased the bioactive compounds extractability with steam oven and sous-vide displaying similar effects, lower than boiling. In boiled Cheddar cauliflower, the total carotenoids and tocopherols contents increased with cooking time until they were 13-fold and 6-fold more than in raw cauliflower, respectively. Conversely, in the Depurple variety, contents increased by half with respect to the orange variety. However, from a nutritional point of view, no differences were revealed among the three different cooking treatments in terms of vitamin A and E levels expressed in μg/100 g of fresh vegetable because of the higher water content of boiled samples that must be considered when evaluating the effect of thermal treatment on cauliflower nutritional traits.

Microalgae can use either ammonium or nitrate for its growth and vitality. However, at a certain level of concentration, ammonium nitrogen exhibits toxicity which consequently can inhibit microalgae productivity. Therefore, this study is aimed to investigate the tolerance of Tetraselmis tetrathele to high ammonium nitrogen concentrations and its effects on growth rate, photosynthetic efficiency (Fv/Fm), pigment contents (chlorophyll a, lutein, neoxanthin, and β-carotene), and fatty acids production. Experiments were performed at different ammonium nitrogen concentrations (0.31–0.87 gL−1) for 6 days under a light source with an intensity of 300 μmol photons m−2 s−1 and nitrate-nitrogen source as the experimental control. The findings indicated no apparent enhancement of photosynthetic efficiency (Fv/Fm) at high levels of ammonium nitrogen (NH4+-N) for T. tetrathele within 24 h. However, after 24 h, the photosynthetic efficiency of T. tetrathele increased significantly (p < 0.05) in high concentration of NH4+-N. Chlorophyll a content in T. tetrathele grown in all of the different NH4+-N levels increased significantly compared to nitrate-nitrogen (NO3-N) treatment (p < 0.05); which supported that this microalgal could grow even in high level of NH4+-N concentrations. The findings also indicated that T. tetrathele is highly resistant to high ammonium nitrogen which suggests T. tetrathele to be used in the aquaculture industry for bioremediation purpose to remove ammonium nitrogen, thus reducing the production cost while improving the water quality.

The aim of this study was the use and revalorization of two persimmon by-products A and B generated in the juice production process. The by-product B resulting from a pectinase enzymatic treatment of peels and pulp to optimize juice extraction was especially suitable for recovery of valuable bioactive carotenoids. The extraction solvents and solvent combinations used were: ethanol, acetone, ethanol/acetone (50:50 v/v) and ethanol/acetone/hexane (25:25:50 v/v/v). HPLC-DAD analysis detected and identified a total of nine individual carotenoids namely violaxanthin, neoxanthin, antheraxanthin, lutein, zeaxanthin, β-cryptoxanthin 5,6-epoxide, β-cryptoxanthin, α-carotene, and β-carotene. β-cryptoxanthin and β-carotene represented 49.2% and 13.2% of the total carotenoid content (TCC) in the acetone extract from by-product B. TCC contributed greatly to antioxidant activity of acetone extract derived from this by-product. Pectinase enzymatic treatment of persimmon peels and pulp followed by absolute acetone extraction of carotenoids could be an efficient method to obtain a rich extract in these compounds that could be used as nutraceutical ingredient., This study was supported by Ministerio de Ciencia, Innovación y Universidades through the funded project ‘Simbiosis industrial en el aprovechamiento integral del caqui (Diospyros kaki); Ejemplo de bioeconomía’ (CTM2017-88978-R). Sara Gea-Botella thanks the Agencia Estatal de Investigación la Ayuda para la Formación de Doctores en Empresas “Doctorados Industriales“ (DI-16-08465) through the R+D+i project entitled ‘Evaluación in vitro e in vivo de un extracto procedente de subproductos de la industrialización del caqui’. The authors wish to thank Mitra Sol Technologies S.L. the given technical assistance.

This study provides new insights into the role of photoprotection in preformed and induced defence of two wheat genotypes with contrasting phenotypes to infection by Zymoseptoria tritici. We investigated the mechanisms of the photoprotective response during early infection, including nonphotochemical quenching (NPQ), β-carotene-derived xanthophylls, reactive oxygen species, and the phytohormones abscisic acid (ABA), jasmonic acid (JA), and salicylic acid (SA). Furthermore, we quantified the effects of pathogenesis on photosynthesis, stomatal control, and expression of plant defence molecular markers. The photoprotective mechanism of successful defence involved the qI component of NPQ leading to rapid down-regulation of photosystem II quantum yield and chlorophyll a:b, increased biosynthesis of the xanthophyll neoxanthin and ABA, and the expression of chloroplast-specific enzymes to engage in scavenging of O. Elevated ABA in the resistant genotype correlated with preformed leaf defence traits including low stomatal density, increased expression of wax biosynthesis, and lignification. Z. tritici exhibited reduced germination and branching on the resistant host genotype and hijacked stomatal control in both genotypes by enhancing stomatal sensitivity to light. Increased biosynthesis of JA and anthocyanins, in contrast to SA, were quantified in the incompatible interaction. Our results indicate that ABA and JA in antagonistic action to SA were associated with defence in the resistant genotype, Cougar, against Z. tritici., This work was carried out under the Capture project supported by Innovate UK (grant no. 38936-273309) and the Biotechnology and Biological Sciences Research Council (grant no. BB/M011550/1), led by Agrii in collaboration with RAGT Seeds

Carotenoids are important coloration molecules and indispensable component of the human diet. And these compounds confer most of the apricot fruit yellow or orange color. In China, fruit of some apricot cultivar present light-yellow color but strong flowery flavor, however, the chemical mechanism remains unknown. Here, carotenoids and aroma volatile apocarotenoids (AVAs) in three skin types of apricot cultivars (orange, yellow, and light-yellow skinned) were determined by HPLC and GC-MS, respectively. And the transcript levels of carotenogenic genes were analyzed by qRT-PCR. The orange-skinned cultivars “Hongyu” and “Danxing” fruit presented the most abundant total carotenoid, β-carotene and specific α-carotene contents, and β-carotene (52–77%) increased to become the dominant carotenoid during fruit ripening. The transcript levels of lycopene β-cyclase (LCYb) and β-carotene hydroxylase (CHYb) sharply increased during ripening. The yellow-skinned cultivars “Sulian No. 2” and “Akeyaleke” fruit contained lower levels of total carotenoids and β-carotene but were rich in phytoene. The light-yellow coloration of “Baixing” and “Luntaixiaobaixing” fruit was attributed to low amounts of total carotenoids, lutein, and neoxanthin and an absence of β-cryptoxanthin, but high level of aroma volatile apocarotenoids (AVAs) such as β-ionone were detected in these cultivars fruit, accompanied by low transcript levels of carotene hydroxylase (CYP) and zeaxanthin epoxidase (ZEP) but high levels of carotenoid cleavage dioxygenase 1 (CCD1) and CCD4. Correlation analysis showed that the expression level of CCD1 negatively correlated with carotenoid accumulation but positively with AVAs production. These collected results suggest that both carotenoid biosynthesis and degradation are important for apricot coloration and aroma formation. CYP, ZEP, CCD1, and CCD4 may be the key regulation points for carotenoid and AVAs accumulation in apricot fruit, which provide important targets for quality-oriented molecular breeding.

As the periodic emission of light pulses by light emitting diodes (LEDs) is known to stimulate growth or induce high value biocompounds in microalgae, this flashing light regime was tested on growth and biochemical composition of the microalgae Nannochloropsis gaditana, Koliella antarctica and Tetraselmis chui. At low flashing light frequencies (e.g., 5 and 50 Hz, Duty cycle = 0.05), a strain-dependent growth inhibition and an accumulation of protein, polyunsaturated fatty acids, chlorophyll or carotenoids (lutein, β-carotene, violaxanthin and neoxanthin) was observed. In addition, a 4-day application of low-frequency flashing light to concentrated cultures increased productivities of eicosapentaenoic acid (EPA) and specific carotenoids up to three-fold compared to continuous or high frequency flashing light (500 Hz, Duty cycle = 0.05). Therefore, applying low-frequency flashing light as finishing step in industrial production can increase protein, polyunsaturated fatty acids or pigment contents in biomass, leading to high-value algal products.

Choy sum is a commonly consumed Asian green leafy brassica vegetable. A comprehensive spectrum of nutritional important metabolites, including amino acids, plant sugars, essential minerals, vitamins (A, B9, E, and K1) and glucosinolates were systematically quantified using LC-QQQ-MS, GC-QQQ-MS and ICP-MS. Significant metabolic profile shifts were observed during the three major developmental stages (microgreen, seedling and adult) studied. Primary metabolites, especially essential amino acids decreased while most plant sugars increased from microgreens to seedlings. Carotenoids, such as violaxanthin, neoxanthin, together with vitamin K1 were higher in the seedlings whereas CHO-folate vitamers and β-cryptoxanthin were much lower in adult plants. Most essential minerals were concentrated in the microgreens, while sodium increased in adult plants. Aliphatic glucosinolates in microgreens were converted to indolic glucosinolates in the seedlings and further to aromatic glucosinolates in the adults. Overall findings reveal that most of the nutritional metabolites were concentrated either in the microgreens or seedlings.

Light is a paramount parameter driving photosynthesis. However, excessive irradiance leads to the formation of reactive oxygen species that cause cell damage and hamper the growth of photosynthetic organisms. Xanthophylls are key pigments involved in the photoprotective response of plants and algae to excessive light. Of particular relevance is the operation of xanthophyll cycles (XC) leading to the formation of de-epoxidized molecules with energy dissipating capacities. Neoxanthin, found in plants and algae in two different isomeric forms, is involved in the light stress response at different levels. This xanthophyll is not directly involved in XCs and the molecular mechanisms behind its photoprotective activity are yet to be fully resolved. This review comprehensively addresses the photoprotective role of 9′-cis-neoxanthin, the most abundant neoxanthin isomer, and one of the major xanthophyll components in plants’ photosystems. The light-dependent accumulation of all-trans-neoxanthin in photosynthetic cells was identified exclusively in algae of the order Bryopsidales (Chlorophyta), that lack a functional XC. A putative photoprotective model involving all-trans-neoxanthin is discussed.

International audience; Abscisic acid (ABA), a plant hormone synthesized from carotenoids, functions in seed germination and abiotic stress responses. ABA is derived from the cleavage of 9-cis-isomers of violaxanthin and neoxanthin, which are oxygenated carotenoids, also called xanthophylls. Although genes encoding enzymes responsible for most steps of the ABA biosynthesis pathway have been identified, enzymatic reactions leading to the production of these cis-isomers from trans-violaxanthin remain poorly understood. Two mutants that lack trans- and cis-neoxanthin, tomato (Solanum lycopersicum) neoxanthin-deficient1 (nxd1) and Arabidopsis (Arabidopsis thaliana) ABA-deficient4 (aba4), were identified previously, but only aba4 exhibited ABA-deficient phenotypes. No enzymatic activity was detected for ABA4 and NXD1 proteins, and their exact function remained unknown. To further investigate ABA4 and NXD1 function in Arabidopsis, we compared phenotypes of single and double mutants, and analyzed the effect of ABA4 overexpression on ABA and carotenoid accumulation in wild-type and mutant backgrounds. We provide convergent evidence that ABA4 is not only required for the formation of trans- and 9 '-cis-neoxanthin from trans-violaxanthin, but also controls 9-cis-violaxanthin accumulation. While nxd1 produces high amounts of 9-cis-violaxanthin and ABA, aba4 nxd1 exhibits reduced levels in both leaves and seeds. Furthermore, ABA4 constitutive expression in nxd1 increases both 9-cis-violaxanthin and ABA accumulation. Subcellular localization of NXD1 protein in transient expression assays suggests that production of the NXD1-derived factor required for neoxanthin synthesis takes place in the cytosol. Finally, we postulate that ABA4, with additional unknown cofactor(s), is required for, or contributes to, trans-to-cis violaxanthin isomerase activity, producing both cis-xanthophyll precursors of ABA.

In many viticulture regions, multiple summer stresses are occurring with increased frequency and severity because of warming trends. Kaolin-based particle film technology is a technique that can mitigate the negative effects of intense and/or prolonged drought on grapevine physiology. Although a primary mechanism of action of kaolin is the increase of radiation reflection, some indirect effects are the protection of canopy functionality and faster stress recovery by abscisic acid (ABA) regulation. The physiological mechanism underlying the kaolin regulation of canopy functionality under water deficit is still poorly understood. In a dry-down experiment carried out on grapevines, at the peak of stress and when control vines zeroed whole-canopy net CO2 exchange rates/leaf area (NCER/LA), kaolin-treated vines maintained positive NCER/LA (~2 &micro, mol m&minus, 2 s&minus, 1) and canopy transpiration (E) (0.57 &micro, 1). Kaolin-coated leaves had a higher violaxanthin (Vx) + antheraxanthin (Ax) + zeaxanthin (Zx) pool and a significantly lower neoxanthin (Nx) content (VAZ) when water deficit became severe. At the peak of water shortage, leaf ABA suddenly increased by 4-fold in control vines, whereas in kaolin-coated leaves the variation of ABA content was limited. Overall, kaolin prevented the biosynthesis of ABA by avoiding the deviation of the VAZ epoxidation/de-epoxidation cycle into the ABA precursor (i.e., Nx) biosynthetic direction. The preservation of the active VAZ cycle and transpiration led to an improved dissipation of exceeding electrons, explaining the higher resilience of canopy functionality expressed by canopies sprayed by kaolin. These results point out the interaction of kaolin with the regulation of the VAZ cycle and the active mechanism of stomatal conductance regulation.

The commercial cultivation of microalgae began in the 1960s and Chlorella was one of the first target organisms. The species has long been considered a potential source of renewable energy, an alternative for phytoremediation, and more recently, as a growth and immune stimulant. However, Chlorella vulgaris, which is one of the most studied microalga, has never been comprehensively profiled chemically. In the present study, comprehensive profiling of the Chlorella vulgaris metabolome grown under normal culture conditions was carried out, employing tandem LC-MS/MS to profile the ethanolic extract and GC-MS for fatty acid analysis. The fatty acid profile of C. vulgaris was shown to be rich in omega-6, -7, -9, and -13 fatty acids, with omega-6 being the highest, representing more than sixty percent (>60%) of the total fatty acids. This is a clear indication that this species of Chlorella could serve as a good source of nutrition when incorporated in diets. The profile also showed that the main fatty acid composition was that of C16-C18 (>92%), suggesting that it might be a potential candidate for biodiesel production. LC-MS/MS analysis revealed carotenoid constituents comprising violaxanthin, neoxanthin, lutein, β-carotene, vulgaxanthin I, astaxanthin, and antheraxanthin, along with other pigments such as the chlorophylls. In addition to these, amino acids, vitamins, and simple sugars were also profiled, and through mass spectrometry-based molecular networking, 48 phospholipids were putatively identified.

In this work, we studied the effectiveness of using spontaneous and resonant Raman spectroscopy of blood plasma of patients as an express diagnosis of preeclampsia. As the resonant wavelength, radiation with a wavelength of 532 nm was chosen, which is characteristic of the excitation of a signal of carotenoids. It was found that the contribution of certain types of carotenoids (β – carotene, astaxanthin, mixoxanthophyll, lycopene, deinoxanthin, spirilloxanthin, neoxanthin, fucoxanthin) significantly changes during the development of the disease and it was noted that the total concentration of all types of carotenoids in blood plasma is normally significantly higher than with preeclampsia.

Ecologically relevant low UV-B is reported to alter reactive oxygen species metabolism and anti-oxidative systems through an up-regulation of enzymes of the phenylpropanoid pathway. However, little is known about low UV-B-induced changes in carotenoid profile and their impacts on light harvesting and photoprotection of photosystem II (PSII) in plants. We investigated carotenoids profile, chlorophyll pigments, phenolics, photosynthetic efficiency and growth in Arabidopsis thaliana (Col-0) plants grown under photosynthetically active radiation (PAR), PAR+ ultraviolet (UV)-A and PAR+UV-A+B regimes for 10 days in order to assess plant acclimation to low UV-B radiation. A chlorophyll fluorescence assay was used to examine UV-B tolerance in plants further exposed to acute high UV-B for 4 and 6 h following a 10-day growth under different PAR and UV regimes. We found that both PAR+ UV-A and PAR+UV-A+B regimes had no negative effect on quantum efficiency, electron transport rate, rosette diameter, relative growth rate and shoot dry weight of plants. Chronic PAR+ UV-A regime considerably (P < 0.05) increased violaxanthin (26 %) and neoxanthin (92 %) content in plants. Plant exposure to chronic PAR+UV-A+B significantly (P < 0.05) increased violaxanthin (48 %), neoxanthin (63 %), lutein (33 %), 9-cis s-carotene (28 %), total s-carotene (29 %) and total phenolics (108 %). The maximum photochemical efficiency (Fv/Fm) in leaves was found to be positively correlated with total phenolics (rho = 0.81 and rho = 0.91, P < 0.05 for 4 and 6 h, respectively) and non-photochemical quenching (qN) (rho = 0.81 and rho = 0.84, P < 0.05 for 4 and 6 h, respectively) in plants exposed to acute high UV-B for 4 and 6 h following a 10-day growth under chronic PAR+UV-A+B. There was also a significant positive correlation (rho = 0.93, P < 0.01) between qN and lutein content in the plants exposed to acute high UV-B stress for 4 h following plant exposure to chronic PAR+UV-A+B. The findings from our study indicate that plants grown under chronic PAR+UV-A+B displayed higher photoprotection of PSII against acute high UV-B stress than those grown under PAR and PAR+ UV-A regimes. An induction of phenolics and lutein-mediated development of qN were involved in the photoprotection of PSII against UV-B-induced oxidative stress.

The selected A. tricolor accessions contained abundant color attributes, betacyanin, carotenoids, betalains, betaxanthins, and antioxidants potentiality that varied in terms of genotypes. For the first time, we identified 4 betacyanins, and 5 carotenoid compounds in A. tricolor genotypes. The genotype VA14 and VA16 had abundant color attributes, betacyanin such as amaranthine, iso-amaranthine, betanin, iso-betanin, and antioxidants potentiality. These two genotypes having an excellent source of color attributes, betacyanins, betalains, betaxanthins, and antioxidants potentiality could be used as potent antioxidant varieties. The genotype VA11 and VA16 had abundant carotenoid components, such as zeaxanthin, lutein, violaxanthin, neoxanthin, total xanthophylls, and beta-carotene. The genotype VA11 and VA16 had abundant carotenoid components that could be used as carotenoid enrich varieties. It revealed from the correlation study that pigment profiles of A. tricolor genotypes exhibited high quenching capacity of radicals. These accessions have high antioxidant potentials and great opportunity to make drinks, preservatives, and colorant of food products to feed the community deficient in antioxidants. The identified components of betacyanins and carotenoids in A. tricolor require comprehensive pharmacological study. The baseline data on color attributes, betacyanins profile, carotenoids profile, betaxanthins, betalains and antioxidant potentiality obtained in the present study could contribute to pharmacologists for evaluating these components scientifically in A. tricolor.

Nephroselmis sp. was previously identified as a species of interest for its antioxidant properties owing to its high carotenoid content. In addition, nitrogen availability can impact biomass and specific metabolites&rsquo, production of microalgae. To optimize parameters of antioxidant production, Nephroselmis sp. was cultivated in batch and continuous culture conditions in stirred closed photobioreactors under different nitrogen conditions (N-repletion, N-limitation, and N-starvation). The aim was to determine the influence of nitrogen availability on the peroxyl radical scavenging activity (oxygen radical absorbance capacity (ORAC) assay) and carotenoid content of Nephroselmis sp. Pigment analysis revealed a specific and unusual photosynthetic system with siphonaxanthin-type light harvesting complexes found in primitive green algae, but also high lutein content and xanthophyll cycle pigments (i.e., violaxanthin, antheraxanthin, and zeaxanthin), as observed in most advanced chlorophytes. The results indicated that N-replete conditions enhance carotenoid biosynthesis, which would correspond to a higher antioxidant capacity measured in Nephroselmis sp. Indeed, peroxyl radical scavenging activity and total carotenoids were higher under N-replete conditions and decreased sharply under N-limitation or starvation conditions. Considering individual carotenoids, siphonaxanthin, neoxanthin, xanthophyll cycle pigments, and lycopene followed the same trend as total carotenoids, while &beta, carotene and lutein stayed stable regardless of the nitrogen availability. Carotenoid productivities were also higher under N-replete treatment. The peroxyl radical scavenging activity measured with ORAC assay (63.6 to 154.9 &micro, mol TE g&minus, 1 DW) and the lutein content (5.22 to 7.97 mg g&minus, 1 DW) were within the upper ranges of values reported previously for other microalgae. Furthermore, contents of siphonaxanthin ere 6 to 20% higher than in previous identified sources (siphonous green algae). These results highlight the potential of Nephroselmis sp. as a source of natural antioxidant and as a pigment of interest.

Three wild edible plant species belonging to the Asteraceae family, Crepis vesicaria L. (s.l.), Sonchus asper (L.) Hill s.l., and Sonchus oleraceus L., usually consumed in the Mediterranean diet, were tested for their nutritional composition and content of carotenoids, tocols, thiamine and riboflavin. Low amounts of thiamine and riboflavin were found. All species were sources of xanthophylls (violaxanthin, neoxanthin, lutein, zeaxanthin and β-cryptoxanthin) and carotenes (α-carotene, β-carotene, 9-cis-β-carotene and 13-cis-β-carotene). Lutein accounted for the highest content (about 4mg/100g). They had good tocol amounts, in particularα-tocopherol (about 2–3mg/100g). Taking into account the Recommended Daily Allowance (RDA) established by the EU Regulation, the analyzed plants can be declared as a source of fiber, vitamin A and E. These data could be useful for database on the nutritional and bioactive compound profile of studied plants and can contribute in promoting their use in functional foods.

Carotenoids participate in photosynthesis and photoprotection in oxygenic phototrophs. Euglena gracilis, a eukaryotic phytoflagellate, synthesizes several carotenoids: β-carotene, neoxanthin, diadinoxanthin and diatoxanthin. Temperature is one of the most striking external stimuli altering carotenoid production. In the present study, to elucidate the regulation of carotenoid synthesis of E. gracilis in response to environmental stimuli, we functionally identified phytoene desaturase genes (crtP1 and crtP2) and the ζ-carotene desaturase gene (crtQ) of this alga and analyzed expression of those genes and the composition of major carotenoids in cells grown under cold (20�C) and high-intensity light (HL; 240 �mol photon m-2 s-1) conditions. 20�C-HL treatment increased the transcriptional level of the phytoene synthase gene (crtB), and crtP1 and crtP2, whose products catalyze the early steps of carotenoid biosynthesis in this alga. Cultivation at 20�C under illumination at 55 �mol photon m-2 s-1 (low-intensity light; LL) decreased the cell concentration, Chl and total major carotenoid content by 61, 75 and 50%, respectively, relative to control (25�C-LL) cells. When grown at 20�C-HL, the cells showed a greater decrease in cell concentration and photosynthetic pigment contents than those in 20�C-LL. β-Carotene, neoxanthin and diadinoxanthin contents were decreased by more than half in 20�C-LL and 20�C-HL treatments. On the other hand, when subjected to 20�C-LL and 20�C-HL, the cells retained a diatoxanthin content comparable with control cells. Our findings suggested that diatoxanthin plays crucial roles in the acclimation to cold and intense light condition. To the best of our knowledge, this is the first report on a photosynthetic organism possessing dual crtP genes.

The present study deals with optimization of alkali treatment including KOH concentration (20, 30, 40 and 50% w/v), treatment time (0, 10, 20, 40, 60 and 120 min) and temperature (30, 50, 70 and 90 °C) in order to eliminate the green pigments (chlorophyll residues) of lutein acetone extract from spinach. Based on spectrophotometric and RP-HPLC-PDA analyses, alkali treatment (KOH 30% w/v) at 50 °C for 40 min with 4:1 mass ratio (extract: alkali solution) were the optimum conditions to eliminate chlorophyll residues of spinach leaf powder (extracted with acetone for 12 h). As a result, the lutein content of the alkali treated extract was measurable (1.85 mg/g dry weight basis), whereas its quantification was unlikely due to the interferences of green and orange pigments. Regarding the lutein content of marigold petals (14.48 mg/g dry weight basis) extracted by acetone for 6 h, it was noticed that heat treatment alone (50 °C for 40 min without KOH) was ineffective, while by combination of alkali and temperature (KOH 30% w/v, for 40 min at 25, 50 or 70 °C) the lutein content was significantly (p

Selenium (Se) is an essential trace element for human nutrition and a key component of selenoproteins having fundamental biological and nutraceutical functions. We currently examined lettuce biofortification with Se in an open-gas-exchange growth chamber using closed soilless cultivation for delivering Se-rich food. Morphometric traits, minerals, phenolic acids, and carotenoids of two differently pigmented Salanova cultivars were evaluated in response to six Se concentrations (0–40 μM) delivered as sodium selenate in the nutrient solution. All treatments reduced green lettuce fresh yield slightly (9%), while a decrease in red lettuce was observed only at 32 and 40 μM Se (11 and 21% respectively). Leaf Se content increased in both cultivars, with the red accumulating 57% more Se than the green. At 16 μM Se all detected phenolic acids increased, moreover a substantial increase in anthocyanins (184%) was recorded in red Salanova. Selenium applications slightly reduced the carotenoids content of green Salanova, whereas in red Salanova treated with 32 μM Se violaxanthin + neoxanthin, lutein and β-cryptoxanthin spiked by 38.6, 27.4, and 23.1%, respectively. Lettuce constitutes an ideal target crop for selenium biofortification and closed soilless cultivation comprises an effective tool for producing Se-enriched foods of high nutraceutical value.

In this article matrix components in spinach were investigated in detail. The samples were prepared using two QuEChERS (quick, easy, cheap, effective, rugged and safe) methods, AOAC and CEN. Liquid chromatography–electrospray ionization–tandem mass spectrometry (LC–ESI–MS/MS), gas chromatography–mass spectrometry (GC–MS) and ultra performance liquid chromatography-diode array detector (UPLC-DAD), were applied for component identification. The strategies of identification by LC–ESI–MS/MS include accurate mass spectra of the parent ion, comparison with previous literature data and investigation of the mass spectral decomposition pattern. Overall, fourteen components were identified by LC–ESI–MS/MS in each methods of AOAC and CEN, which were phytosteroids, flavonoids, fatty acids and fatty acid amides. Fifty components using AOAC method and fifty-seven components using CEN method were identified in GC–MS by comparing mass data with the National Institute of Standards and Technology (NIST, U.S.) database. The results indicate that the major components of the matrix are terpenoids, fatty acids and fatty acid esters. Moreover, three pigments (neoxanthin, violaxanthin and lutein) in the AOAC method and eight pigments (neoxanthin, violaxanthin, zeaxanthin, lutein, chlorophyll a, chlorophyll b, pheophytin a and beta-carotene) in the CEN method that gave a characteristics peak at 440 nm were identified by the UPLC-DAD. According to the sample preparation condition using different amounts of graphitized carbon black (GCB) in this study, the AOAC method had higher matrix component removal efficiency than the CEN method. A better understanding of matrix components would increase the current knowledge for improvement of existing QuEChERS methodology, as well as contribute to new method developments. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s13197-018-3318-4) contains supplementary material, which is available to authorized users.

Regional climate change in Antarctica would favor the carbon assimilation of Antarctic vascular plants, since rising temperatures are approaching their photosynthetic optimum (10-19°C). This could be detrimental for photoprotection mechanisms, mainly those associated with thermal dissipation, making plants more susceptible to eventual drought predicted by climate change models. With the purpose to study the effect of temperature and water availability on light energy utilization and putative adjustments in photoprotective mechanisms of Deschampsia antarctica Desv., plants were collected from two Antarctic provenances: King George Island and Lagotellerie Island. Plants were cultivated at 5, 10 and 16°C under well-watered (WW) and water-deficit (WD, at 35% of the field capacity) conditions. Chlorophyll fluorescence, pigment content and de-epoxidation state were evaluated. Regardless of provenances, D. antarctica showed similar morphological, biochemical and functional responses to growth temperature. Higher temperature triggered an increase in photochemical activity (i.e. electron transport rate and photochemical quenching), and a decrease in thermal dissipation capacity (i.e. lower xanthophyll pool, Chl a/b and β carotene/neoxanthin ratios). Leaf mass per unit area was reduced at higher temperature, and was only affected in plants exposed to WD at 16°C and exhibiting lower electron transport rate and amount of chlorophylls. D. antarctica is adapted to frequent freezing events, which may induce a form of physiological water stress. Photoprotective responses observed under WD contribute to maintain a stable photochemical activity. Thus, it is possible that short-term temperature increases could favor the photochemical activity of this species. However, long-term effects will depend on the magnitude of changes and the plant's ability to adjust to new growth temperature.

In order to investigate the regulation mechanisms of carotenoid biosynthesis in mango flesh, carotenoid content and the expression patterns of 15 carotenogenic gene in flesh of two mango cultivars ‘Tainong1’and ‘Hongyu’ were analsyed during fruit development and ripening. Carotenoid were increasingly accumulated during fruit growth and sharply incaresed during postharvest ripening in both cultivars. In ripe fruit, total carotenoid content in the flesh of ‘Tainong1’ was 177.16 ug/g (Fresh weight) FW, and for ‘Hongyu’ 36.24 ug/gFW. The major carotenoids in the flesh were β-Carotene and α-carotene,with minor carotenoids being β-cryptoxanthin, zeaxanthin, lutin and neoxanthin. RT-PCR analysis revealed that the expression of carotenogenic genes CRTISO, PSY, ZDS, BCH and ZEP were up-regulated in flesh, whereas the transcript levels of LCYB, LCYE and NCED were down-regulated in two cultivars and during fruit development and ripening. CRTISO, PSY, ZDS, BCH and ZEP transcripts were significantly positively correlated with the total carotenoids content, while no differences were detected for CRTISO expression between cultivars. These results suggested that PSY, ZDS, BCH and ZEP coordinately contribute to carotenoid accumulation during mango fruit development and ripening.

The chemical and biological properties of carotenoids in the freshwater alga Oedogonium intermedium were investigated in this study. Carotenoids were extracted from the alga by dichloromethane and purified by saponification. The carotenoid content was determined both spectrometrically and by HPLC, the carotenoids identified by HPLC-PDA-APCI-IT-TOF-MS and the extracts analysed for several health-related bioactivities. The crude and saponified extracts contained 3,411.2±20.7 and 2,929.6±5.9µg carotenoids/g dry algal biomass, respectively. Seven major carotenoids were identified, namely neoxanthin, 9'-cis-neoxanthin, loroxanthin, violaxanthin, lutein, α-carotene and β-carotene, which were present in similar amounts in the alga. Both the crude and saponified carotenoid extracts exhibited significant antioxidant activities as well as potent inhibitory effects against several metabolically important enzymes including α-amylase, α-glucosidase, pancreatic lipase and hyaluronidase, but they were poor inhibitors of angiotensin converting enzyme (ACE). Oedogonium could be an important new source of carotenoids, specifically loroxanthin, which is lacking in terrestrial plants.