Higher Carotenoid Research Articles

Grain yellowness is an effective predictor of carotenoid content in global sorghum populations

Identification of high carotenoid germplasm is crucial to assist breeders in provitamin-A biofortification of sorghum (Sorghum bicolor [L.] Moench). High-performance liquid chromatography is the gold standard for carotenoid quantification, however, it is not feasible for large scale phenotyping due to its high cost and low throughput. In this study, we tested the feasibility of using grain color as a high-throughput method of carotenoid biofortification breeding. We hypothesized that visual, color-based selection can be an effective strategy to identify high-carotenoid accessions. Yellow grain had significantly higher carotenoid content than red, brown, and white grain. The degree of yellowness could distinguish the presence or absence of carotenoids, but could not distinguish carotenoid concentrations within yellow-only accessions. The degree of luminosity of the grain, however, was able to better predict carotenoid concentrations within yellow-only accessions. Genome-wide association studies identified significant marker-trait associations for qualitative and quantitative grain color traits and carotenoid concentrations near carotenoid pathway genes—ZEP, PDS, CYP97A, NCED, CCD, and LycE—three of which were common between grain color and carotenoid traits. These findings suggest that using grain color as a method for screening germplasm may be an effective high-throughput selection tool for prebreeding and early-stage breeding in carotenoid biofortification.

Supercritical CO2 as a green technology for carotenoids-rich virgin palm oil production: Process optimization, kinetics and thermodynamics modeling

The experimental conditions of supercritical carbon dioxide (scCO2) extraction of virgin palm oil (VPO) from oil palm mesocarp fiber (OPMF) were optimized using the Response Surface Methodology (RSM). Results showed that a maximum of 28.68% of VPO extraction was obtained at the optimal experimental conditions of scCO2 extraction: pressure of 31MPa, temperature of 340K, and extraction time of 80min. The second-order rate equation, Arrhenius equation and Eyring theory were employed to assess the kinetics behaviour, activation energy and thermodynamics behaviour of scCO2 extraction of VPO from oil palm mesocarp fiber. The lower activation energy value (12.17kJ/mol) of scCO2 for the extraction of VPO from OPMF indicates that the scCO2 extraction technology is less dependent on temperature during the extraction of VPO from OPMF. The physicochemical properties and fatty acids compositions analyses reveal that the scCO2 extracted VPO contains high carotenoids content (982μg/g), low free fatty acids content (0.31wt.%), higher oxidative stability and higher unsaturated fatty acids content.

Establishment of near-infrared rapid prediction model and comprehensive evaluation model for foxtail millet quality

Foxtail millet (Setaria italica) is one of the earliest cereal crops in the world and is still an important food crop in Asia, Europe and Africa. However, there are few reports on rapid and comprehensive evaluations of foxtail millet quality. In this study, color, nutrient content, cooking quality and sensory quality of 45 varieties of foxtail millet from the main production areas of the country were determined. Near infrared spectroscopy (NIRS) combined with partial least squares regression (PLSR) was developed for rapid and nondestructive detection of foxtail millet quality, where NIRS models for ash, amylopectin, protein, and VB1 and VB2 were desirable. In addition, correlation analysis, principal component analysis and cluster analysis were used for comprehensive evaluation of millet quality. The results showed that foxtail millet with low ash, protein, fat and fiber content and high amylopectin and carotenoid content has good cooking quality and was more preferred. This research will contribute to the quality evaluation and further processing of foxtail millet.

Be prepared: how does discontinuous hydration in Tabebuia heterophylla seeds induce stress tolerance in seedlings?

Discontinuous hydration and dehydration (HD) cycles refer to controlled imbibition followed by dehydration before seed germination. Here, we investigated whether the level of imbibition before HD cycles affects the physiology of Tabebuia heterophylla seeds and seedlings. Seeds were imbibed for 10 h (T1; phase I of imbibition) or 35 h (T2; phase II), dehydrated, and progressively rehydrated one to four times (HD cycles). Germination and biochemical parameters (membrane integrity; total soluble, reducing, and nonreducing (NRS) sugars; proteins, amino acids, proline, H2O2, catalase, ascorbate peroxidase, and glutathione reductase activity) were quantified at the last rehydration step of each cycle. Biometric and biochemical parameters (including pigments) were analysed in seedlings 60 days after germination. HD cycles at T1 led to reduced seed germination and greater plasma membrane damage, higher enzyme activity (catalase and glutathione reductase) and accumulation of NRS, total amino acids, and proline compared to the controls and T2 treatment. Cellular damage became more severe with more HD cycles. HD cycles at T2 synchronized germination regardless of the number of cycles and also had a priming effect. T2 seeds had less NRS, total amino acids, and proline content than T1. HD cycles at T1 produced seedlings with higher carotenoid and total chlorophyll content than controls and T2, while seedlings from HD cycles at T2 had higher amounts of osmoprotectants. HD cycles at T2 benefited seeds and seedlings more than at T1. This suggests that the physiological and biochemical effects of HD cycles in seeds modulate seedling plasticity, depending on water availability, potentially promoting increased tolerance to recurrent droughts that will be intensified with ongoing climate changes.

Eryngo essential oil nanoemulsion stabilized by sonicated-insect protein isolate: An innovative edible coating for strawberry quality and shelf-life extension

New bioactive coatings with eryngo essential oil (EEO) nanoemulsions stabilized by ultrasonically-treated lesser mealworm protein isolate (LMPI) were developed to extend strawberry shelf life and quality. EEO due to high carvone (43.03 %), phenolics (87.45 mg gallic acid equivalent/g), flavonoids (13.56 mg quercetin equivalent/g), and carotenoids (635.07 mg/kg) contents exhibited a significant antioxidant activity comparable to ascorbic acid (AA) and BHT. Nanoemulsions stabilized with 9 % sonicated LMPI showed smaller droplet size, higher negative ζ-potential, and greater stability, turbidity, and encapsulation efficiency of EEO compared to those stabilized with native LMPI. The FTIR spectra showed that sonicated LMPI had structural changes enhancing its emulsifying activity, with key peaks indicating the presence of hydrogen bonds, carbonyl groups, and protein conformations in both EEO and LMPI. Strawberries coated with optimal EEO-loaded nanoemulsions showed superior quality with minimal storage-dependent physicochemical, textural, color, and sensory changes compared to control samples. This edible coating also maintained higher total monomeric anthocyanin and AA contents with lower peroxidase activity during storage than EEO-based coatings. However, no significant difference in superoxide dismutase activity between samples covered by EEO and EEO-loaded nanoemulsions over 14 days of storage was found. Bioactive nanoemulsions stabilized by insect proteins would be an eco-friendly and safe approach to upholding quality standards in stored fruits and vegetables.

Molecular protocol to develop β-carotene-biofortified rice events via molecular optimization

Providing food with nutrition and functionality is crucial for sustaining human life. Rice (Oryza sativa L.) is a representative staple crop with high carbohydrate content but low amounts of essential amino acids, micronutrients, and carotenoids such as provitamin A. To improve the nutritional quality, rice endosperm was biofortified to accumulate carotenoids such as β-carotene through genetic engineering (i.e., using synthetic carotenoid biosynthetic genes, a nonmammalian viral polycistronic sequence, and an optimized promoter and transit peptide) and high-throughput rice transformation (approximately 300 transgenic plants per construct). To facilitate the safety assessment of genetically modified food, molecular characterization was performed to select elite lines equipped with a single intergenic insertion of T-DNA, high transgene expression, in this case leading to high carotenoid content, and with phenotypic and compositional substantial equivalence. In this study, we present β-carotene-biofortified rice event candidate lines eligible for commercial use and a disclosed molecular protocol for the development of biotech rice crops.

Genome-wide association study of carotenoids in maize kernel.

In this study, the contents of four carotenoids in 244 maize inbred lines were detected and about three million single nucleotide polymorphisms (SNPs) for genome-wide association study to preliminarily analyze the genetic mechanism of maize kernel carotenoids. We identified 826 quantitative trait loci (QTLs) were significantly associated with carotenoids contents, and two key candidate genes Zm00001d029526 (CYP18) and Zm00001d023336 (wrky91) were obtained. In addition, we found a germplasm IL78 with higher carotenoids. The results of this study can provide a theoretical basis for screening genes that guide kernel carotenoids selection breeding.

Comparative analysis of carotenoids, fatty acids, minerals, tastes, and odor in the skin of wild versus farmed large yellow croaker: Superior nutritional benefits with elevated heavy metal risks

People commonly consume large yellow croaker with skin, yet comprehensive studies on the skin's nutritional value, flavor, and health risks are lacking. The skin from wild fish and those from five main farming areas, nearly representing all fish in the market, were analyzed. The study revealed that the skin of all tested fish exhibited high levels of carotenoids, EPA, DHA, Σn-3/Σn-6 PUFA, Fe, Se, Cu, Zn, and various flavor compounds including free amino acids, 5′-nucleotides, and odor substances, and low levels of atherosclerotic and thrombogenic indices, suggesting health benefits. Notably, the wild fish skin was yellower with high carotenoid accumulation and nutritionally superior in fatty acids but showed a high heavy metal risk. Additionally, this research has, for the first time, elucidated that carotenoid storage in the skin of large yellow croaker depends on lutein esterification with oleic, palmitic, and decanoic acids. In summary, the current study indicates that the skin of farmed fish offers significant nutritional value and favorable flavor profiles, making it safe for consumption. However, while the skin of wild large yellow croaker is more nutritionally advantageous, it also presents a non-negligible risk of heavy metal contamination.

Effect of different cooking oils and methods on physicochemical, nutritional, and microbial safety for mixed vegetables focusing on Bangladesh

SummaryCooking is an essential aspect of everyday life. Cooking oils and methods exert enormous effects on the nutritional, physicochemical, microbial qualities, and bioactive compounds of vegetables. Most of them are either directly or indirectly related to human health. Hence, the present investigation was undertaken to find out the effect of two cooking methods (traditional and steam cooking) and four selected culinary oils (soybean oil, mustard oil, extra virgin olive oil, and extra virgin coconut oil) on the cooked vegetables. The results indicated that steam‐cooked mixed vegetables retained more nutrients than traditional ones. The traditional cooking method of preparing mixed vegetables with soybean oil revealed higher amounts of crude fat, 26.90 ± 0.10%, whereas steam cooking and traditionally cooked vegetables without oils showed lower amounts of crude fat, 1.21 ± 0.04%, and 1.94 ± 0.04%, respectively. The highest crude fibre from 5.68 ± 0.20 to 6.48 ± 0.02% was recorded by the steam‐cooked vegetables in comparison with traditionally prepared mixed vegetables. Traditionally cooked vegetables with mustard oil showed a higher crude protein (13.30 ± 0.10%). The mixed vegetables that were steam‐cooked using extra virgin olive oil revealed a decreased percent of carbohydrates, 49.42 ± 0.03%. Most of the essential minerals especially for the human body like Ca, Mg, Fe, Cu, and Zn were found notable in both traditional and steam‐cooking processes using extra virgin olive oil, and mustard oil. The leading bioactive compound ß‐carotene 67.04 ± 0.04 mg/100 g was noted by the steam‐cooked vegetables with mustard oil. The highest anthocyanin 32.69 ± 0.02 mg/100 g and ascorbic acid 42.31 ± 0.01 mg/100 g were recorded in the steam‐cooked vegetables without culinary oils. Steam‐cooking with extra virgin coconut and olive oil resulted in high total carotenoid and phenolic content (31.60 ± 0.05 and 3.11 ± 0.04 mg/100 g, respectively). However, the findings suggest that culinary oils used in the present study retained more essential minerals and bioactive compounds than soybean oil. Only sufficient amount of edible oil may be used for vegetables cooking and proper functioning of fat‐soluble vitamins.

Novel avenue of carotenoids & omega-3 rich carrot waste extract in innovative table spread: optimization (based on texture and sensory) and characterization

Carrot waste is a rich source of pro-vitamin A, β-carotene, recognized for its immunomodulatory and health-promoting properties. The present study was aimed at formulating an innovative table spread (ITS) enriched with concentrated carotenoid extract derived from carrot pomace. Prior extraction of carotenoids in flaxseed oil was optimized resulting in extract (CE) rich in carotenoids (82.66 µg/g) and alpha-linolenic acid (ALA). The effects of three ingredients i.e. CE, tri-sodium citrate (TC), and common salt (CS) on ITS formulated using 33 factorial experiment (27 treatment combinations) were determined employing three-way ANOVA of Proc GLM method of SAS 9.3 for its textural and sensory acceptability. The optimized formulation contained 15% CE, 1.0% CS and 0.3% TC. ITS displayed medium (≈51% fat) internal phase emulsion (confirmed via fluorescence microscopy) stabilized (-56.867 mV ± 0.850 zeta potential) by whey protein concentrate and possessed high thermal stability (studied through thermogravimetric analysis). The control table spread (CTS) was prepared using all ingredients except CE replaced with flaxseed oil. ITS showed significantly (p < 0.05) higher carotenoid, antioxidant (ABTS, DPPH and FRAP), yellowness index and b* value than CTS; while non-significant (p > 0.05) difference existed for proximate composition and acidity. ITS turned out to be a naturally colored functional product and excellent source of carotenoids (24.276 μg/g ± 0.076) and ALA (13.816 g/ 100 g fat).

Raman spectroscopic analysis of human blood serum of glaucoma patients supplemented with macular pigment carotenoids.

As all major dietary carotenoids are contained in blood, it is a suitable substrate to evaluate their content, in vivo. Following 18-month supplementation of open-angle glaucoma patients with macula-pigment carotenoids (Lutein, Zeaxanthin and Meso-Zeaxanthin) in the European Nutrition in Glaucoma Management trial, Raman spectroscopic analysis of the carotenoid content of pre- and post-supplementation participant blood serum was carried out, to investigate the systemic impact of the supplementation regimen and explore a more direct way of quantifying this impact using routine blood tests. Using a 532 nm laser source for optimal response, a consistent increase in serum carotenoid concentration was observed in the supplemented serum, highest in patients with initial high baseline carotenoid content. A shift in the 1519 cm-1 carotenoid peak also revealed differences in the carotenoid structural profile of the two groups. The findings highlight the potential of Raman spectroscopy toquantify and differentiate carotenoids directly in blood serum.

Screening of Zea mays L. Germplasm for the High Carotenoid Endosperm using Biochemical and Molecular Approaches

Background: In the rapid growing and busy world, the hidden hunger for the micronutrients, vitamins and protein is increasing on a high rate. Among which retinol is the key vitamin which cannot be synthesized by the humans and need to be supplied in the form of carotenoids. Methods: The present study focused in screening the best maize line with high carotenoid content. Carotenoid content was tested and quantified using HPLC. ε-LCY gene and β-HYD 3 were used to screen the best maize line. Result: Among 8 maize lines tested, 5732 showed high carotenoid content i.e., 24.47 mg/g when tested using conventional UV spectrophotometer studies. Quantification of individual carotenoid component was done by HPLC which showed lutein 7.548 mg/g, zeaxanthin 3.21 mg/g, b-cryptoxanthin 5.691 mg/g and b- carotene 8.455 mg/g were recorded in 5732 maize cultivars. Molecular screening using ε-LCY gene and β-HYD 3 revealed 5732 as the best carotenoid maize line with high carotenoid favorable alleles which can be further used for the inbreeding programs.

Skin Carotenoid Levels Are Associated with Demographic Factors, Body Size, and Fruit and Vegetable Intake in the Japanese Population.

This study aimed to determine the association between demographic factors, body size, and fruit and vegetable intake in the general population, focusing on individuals with both low and high skin carotenoid levels. This cross-sectional study was conducted during the 14th National Convention on the Promotion of Food and Nutrition Education (2019) in Yamanashi, Japan (a rural area) and the Open House 2019 at the National Institute of Biomedical Innovation, Health, and Nutrition in Tokyo, Japan (an urban area). Skin carotenoid measurements were conducted, and the participants were asked to fill out a self-administered questionnaire. The study population consisted of 492 Japanese individuals aged ≥16 years. The odds ratios (ORs) for low skin carotenoid levels were elevated in males, those who were overweight, and those who almost never consumed or consumed only one vegetable dish/day. Conversely, the ORs were lower in those living in Yamanashi, aged 30-39 and ≥70 years, and those who consumed fruit ≥1 time/week. For high skin carotenoid levels, the ORs were higher among those aged ≥70 years, living in Yamanashi, and those who consumed fruit ≥1 time/day or ≥5 vegetable dishes/day. Demographic factors, body size, and habitual fruit and vegetable intake may serve as indicators of skin carotenoid levels.

Peach Palm Fruit (Bactris gasipaes) Peel: A Source of Provitamin A Carotenoids to Develop Emulsion-Based Delivery Systems.

The peach palm fruit (Bactris gasipaes) peel is a byproduct after fruit consumption. The peel flour of two varieties (yellow and red) was separately obtained by hot air drying and was subsequently milled. The proximate analysis showed that the red variety exhibited higher protein, fat, and fiber contents than the yellow one. A higher carotenoid (836.5 ± 24.5 μg/g), phenolic compounds (83.17 ± 1.76 mg GAE/100 g), and provitamin A activity (33.10 ± 0.83 μg retinol/g) were found in the cooked red variety. The carotenoid and phenolic compositions were analyzed by HPLC-PDA-MS, finding β-carotene and γ-carotene to be major compounds. The effect of thermal treatment increased the amount of these provitamin A carotenoids and lycopene, which were detected only in the red variety. Among phenolic compounds, procyanidin dimer (isomer I), feruloyl quinic acid, and several apigenin C-hexosides were identified as major constituents of peach palm epicarp. A carotenoid-rich emulsion-based delivery system was obtained after the optimization (RSM model) of carotenoid extraction with ultraturrax and sunflower oil and further development of an ultrasound-assisted emulsion. The best conditions for a stable emulsion were 73.75% water, 25% carotenoid-rich oil extract, 1.25% emulsifiers, and 480 W of ultrasonic power for 5 min. The optimized emulsion had a total carotenoid content of 67.61 μg/g, Provitamin A activity of 3.23 ± 0.56 μg RAE/g, droplet size of 502.23 nm, polydispersity index of 0.170, and zeta potential of -32.26 mV. This emulsion was chemically and physically stable for 35 days at 30 ± 2 °C, showing potential as a food additive with biofunctional properties. The strategy here developed is an economical and environmentally friendly process that allows the reuse of the byproduct of B. gasipaes.

Assessment of physico-biochemical parameters of Moroccan loquat (&lt;i&gt;Eriobotrya japonica &lt;/i&gt;Lindl.) genotypes using multivariate analysis

The present study was conducted to assess the physico-biochemical variability in 35 genotypes of loquat using multivariate analyses, in order to provide efficient criteria and promising genotypes for the loquat genetic breeding program. Mature fruits were collected from 35 loquat genotypes, belonging to the Zegzel valley, were subjected to physico-biochemical analyses. The results revealed a juice yield ranging from 0.21 to 0.65 g/g FW, and a polyphenols content fluctuating from 322.8 to 366.89 µg GAE. g-1 DM. Regarding DPPH test, with a range of 3.35 and 7.6 µmol TE. g-1 DM, showed a positive correlation with total polyphenol content (r2= 0.76) and flavonoid content (r2= 0.72). Moreover, a negative correlation was recorded between the total soluble solids content and juice yield (r2=- 0.47), which was moderately correlated with vitamin C (r2= 0.59). In addition, the components analysis results revealed a significant and independent contribution of bio-physicochemicalcharacteristics in the loquat variation. Indeed, biochemical characteristics contribute to PC1, while, physicochemical parameters contribute to PC2 and PC3. Furthermore, the hierarchical clustering analysis classified the 35 genotypes into four homogeneous groups independently of their geographic origin. According to these findings, the genotypes T9, T12, Z16, Z17 and TA9 exhibited high total sugar content, while, genotypes T9 and T11 provide high carotenoids, flavonoids, total polyphenols and antioxidant activity level. As results, these genotypes can be directly recommended for the vegetative propagation as well as applied along with the efficient characters in future loquat breeding programs.

Assessment of physico-biochemical parameters of Moroccan loquat (&lt;i&gt;Eriobotrya japonica &lt;/i&gt;Lindl.) genotypes using multivariate analysis

The present study was conducted to assess the physico-biochemical variability in 35 genotypes of loquat using multivariate analyses, in order to provide efficient criteria and promising genotypes for the loquat genetic breeding program. Mature fruits were collected from 35 loquat genotypes, belonging to the Zegzel valley, were subjected to physico-biochemical analyses. The results revealed a juice yield ranging from 0.21 to 0.65 g/g FW, and a polyphenols content fluctuating from 322.8 to 366.89 µg GAE. g-1 DM. Regarding DPPH test, with a range of 3.35 and 7.6 µmol TE. g-1 DM, showed a positive correlation with total polyphenol content (r2= 0.76) and flavonoid content (r2= 0.72). Moreover, a negative correlation was recorded between the total soluble solids content and juice yield (r2=- 0.47), which was moderately correlated with vitamin C (r2= 0.59). In addition, the components analysis results revealed a significant and independent contribution of bio-physicochemicalcharacteristics in the loquat variation. Indeed, biochemical characteristics contribute to PC1, while, physicochemical parameters contribute to PC2 and PC3. Furthermore, the hierarchical clustering analysis classified the 35 genotypes into four homogeneous groups independently of their geographic origin. According to these findings, the genotypes T9, T12, Z16, Z17 and TA9 exhibited high total sugar content, while, genotypes T9 and T11 provide high carotenoids, flavonoids, total polyphenols and antioxidant activity level. As results, these genotypes can be directly recommended for the vegetative propagation as well as applied along with the efficient characters in future loquat breeding programs.

Nutritional composition and anti-nutritional properties of wild edible fruits of northeast India

Forageable fruits are an untapped wealth of nutrients and minerals, and huge numbers are found in the vast wilderness of northeast India, including the hilly state of Mizoram. However, their nutritional properties still need to be explored. Hence, the present study was carried to assess the nutritional and anti-nutritional aspects along with the mineral contents of seven selected wild edible fruits, viz. jackfruit (Artocarpus heterophyllus), rough lemon (Citrus jambhiri), false mangosteen (Garcinia xanthocymus), mulberry (Morus nigra), thereju (Prunus jenkinsiii), wild raspberry (Rubus treutleri) and wild ber (Zizyphus mauritiana) to uncover their immense nutritional values, which could solidify their promising potential for an alternate source of nutrition and increase their economic importance. The research demonstrates that Garcinia xanthocymus contains significant amount of total sugars, reducing sugars, and vitamin E. Artocarpus heterophyllus exhibited higher levels of non-reducing sugar, total soluble solids, chlorophyll, carbohydrate and starch. Citrus jambhiri is rich in moisture, vitamin C, acidity, N, and P. Prunus jenkinsii has high anthocyanin, total carotenoid, K and Mn. Rubus treutleri is a rich source of crude fibre, Mg, Na, Co, Cu, and Fe. Ziziphus mauritiana has higher protein. The Morus nigra has higher fat, energy, Ca, and Zn. These findings suggest that all examined wild edible fruits are rich in vitamins and minerals, which makes them vital in combating malnutrition and highlights their potential for use in the nutraceutical and pharmaceutical sectors in futuristic research works. The outcomes of this present study will provide a database about the nutritional composition, potential health benefits, and economic values of these fruits and raise public consciousness about the conservation and enrichment of biodiversity.

Trellising is advantageous over ground culture for out-of-season, protected production and storage of sweet acorn squash

Acorn squash (Cucurbita pepo) have been a familiar item at produce stands for decades in the United States and Canada, but little known or appreciated elsewhere. Following the breeding and development of sweet-fleshed acorn squash in Israel and its commercial introduction in 2007, acorn squash became a high-priced, popular produce item there. Scarcity of supply in winter has led to attempts to fill the consumer demand by using available protected cultivation infrastructure in the relatively mild area of southwestern Israel for production. Such production has proven feasible but it was not determined whether it would be preferable to allow the plants to simply grow sprawling on the ground or vertically, using trellises to train the plants to grow erect. Two sweet acorn squash hybrid cultivars differing in fruit size were compared, growing on the ground or on trellises, for yield, quality, and storability of the fruits. The hybrids bore fully ripe fruits from December through February, producing 56% higher yields when trellised rather than when allowed to grow on the ground. The fruits of trellised plants of both hybrids were more uniformly black-green and firmer than those of ground-grown plants. Their dry matter content at harvest and after 78 days of storage was very high, averaging 28% and 25%, respectively. Total soluble solids content of the fruit flesh from trellised plants was 19% at harvest and an extraordinary 20% after storage at 10°C, 70% RH. The fruit flesh of trellised plants was also more highly colored and had higher carotenoid, ascorbate, and anti-oxidant contents. Overall, trellising of sweet acorn squash during the winter under protected cultivation resulted in significantly higher yields and the finest fruit quality.

Ultrasound-based strategies for the recovery of microalgal carotenoids: Insights from green extraction methods to UV/MS-based identification

Carotenoids, versatile natural pigments with numerous health benefits, face environmental concerns associated with conventional petrochemical-based extraction methods and limitations of their synthetic equivalents. In this context, this study aims to introduce eco-friendly approaches using ultrasound-based strategies (probe and bath) for the extraction of carotenoids from microalgae, initially focusing on Microchloropsis gaditana and subsequently evaluating the versatility of the method by applying it to other microalgae species of interest (Tisochrysis lutea, Porphyridium cruentum, and Phaeodactylum tricornutum) and defatted microalgal residues. Among the approaches evaluated, the 5-min ultrasonic probe system with ethanol showed comparable carotenoid recovery efficiency to the reference method (agitation, 24 h, acetone) (9.4 ± 2.5 and 9.6 ± 3.2 mg g−1 carotenoids per dry biomass, for the green and the reference method, respectively). Moreover, the method's sustainability was demonstrated using the AGREEprep™ software (scored 0.62 out of 1), compared to the traditional method (0.22 out of 1). The developed method yielded high carotenoid contents across species with diverse cell wall compositions (3.1 ± 0.2, 2.1 ± 0.3, and 4.1 ± 0.1 mg g−1 carotenoid per dry biomass for T. lutea, P. cruentum, and P. tricornutum, respectively). Moreover, the application of the method to defatted biomass showed potential for microalgal valorization with carotenoid recovery rates of 41 %, 60 %, 61 %, and 100 % for M.gaditana, P. tricornutum, T. lutea, and P. cruentum, compared to the original biomass, respectively. Furthermore, by using high-performance liquid chromatography with a diode array detector (HPLC-DAD) and high-resolution mass spectrometry (HRMS), we reported the carotenoid and chlorophyll profiles of the different microalgae and evaluated the impact of the eco-friendly methods. The carotenoid and chlorophyll profiles varied depending on the species, biomass, and method used. In summary, this study advances a green extraction method with improved environmental sustainability and shorter extraction time, underscoring the potential of this approach as a valuable alternative for the extraction of microalgal pigments.

An Insight into the Hormonal Interplay Regulating Pigment Changes and Colour Development in the Peel of ‘Granny Smith’, ‘OPAL®’ and ‘Royal Gala’ Apples

AbstractThree apple cultivars with different peel colour and known ripening dynamics were examined throughout development and on-tree ripening to unravel how the hormonal crosstalk may influence colour changes. Our results indicate that the colour of ripe apples in ‘Granny Smith’ and ‘Opal®’ cultivars was primarily influenced by chlorophyll levels masking the contribution of other pigments, whilst the development of red colour in the ‘Royal Gala’ was not only due to an important loss of chlorophyll during ripening but also to anthocyanins accumulation. The interplay amongst indole-3-acetic acid (IAA), ethylene and abscisic acid (ABA) seemed to control pigment content during the later stages of development, indicating that variations in the hormonal profile contribute to colour differences in ripe apples. In ‘Granny Smith’ apples, basal levels of IAA, ethylene and ABA were associated with the inhibition of chlorophyll loss, which was responsible for its green colour, despite the presence of high carotenoid levels. In contrast, the peel of ‘Opal®’ and ‘Royal Gala’ apples underwent a degreening process facilitated by the IAA-mediated activation of ethylene metabolism, in a crosstalk facilitated by MdARF5. The accumulation of anthocyanins in the peel of ‘Royal Gala’ apples seemed to be regulated in an ethylene-dependent manner, with MdMYB1 acting as the mediator between ethylene signalling and the anthocyanins biosynthesis and vacuolar transport pathways. On the other hand, the decline in carotenoid content during fruit development in the three cultivars appeared to be regulated at the catabolic level through ABA and its derivatives and possibly also facilitated by ethylene-mediated mechanisms. Notably, chlorophyll levels, crucial for the observed colour changes, were likely controlled by MdGLK1, a transcription factor involved in chloroplast biogenesis, working in coordination with ethylene-dependent catabolic enzymes. Overall, understanding the cultivar-specific hormonal regulation of apple colour provides valuable insights for developing preharvest and postharvest strategies to ensure an optimal fruit colour aligned with consumer preferences.