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Search Results (257)

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17 pages, 695 KiB  
Article
Effect of Environmentally Friendly Betalain Extraction Methods on Antioxidant Compounds of Tunisian Opuntia stricta Fruit
by Nadia Smirani, Souhir Bouazizi, Emna Bettaieb, Rachel Torkhani and Moktar Hamdi
Foods 2025, 14(5), 851; https://doi.org/10.3390/foods14050851 - 1 Mar 2025
Viewed by 335
Abstract
This study focuses on the extraction of betalain compounds from Opuntia stricta as a natural alternative to synthetic colorants and sustainable environmentally friendly technology solutions. Non-conventional extraction technologies including microwave (MW) and ultrasound (US) were used alone or in combination. The extraction process [...] Read more.
This study focuses on the extraction of betalain compounds from Opuntia stricta as a natural alternative to synthetic colorants and sustainable environmentally friendly technology solutions. Non-conventional extraction technologies including microwave (MW) and ultrasound (US) were used alone or in combination. The extraction process was conducted for both undried Opuntia stricta (OS) and dried Opuntia stricta (DOS) plant material at two distinct drying temperatures, 40 °C and 60 °C, to assess the stability of betalain molecules. The colorant’s potential was evaluated by determining the betalain content, total phenolic content, and antioxidant activity. The MW (2 min) and MW (2 min) + US (10 min) extraction processes yielded the greatest betalain content in OS fresh weight (FW), with 48.54 ± 0.29 mg/100 g FW and 51.01 ± 0.16 mg/100 g FW, respectively. Furthermore, the results showed a considerable drop in betalain content when the plant material was dried at 40 °C and 60 °C, with reduction rates of 53.75% and 24.82%, respectively, compared to the betalain content before the drying process. The LC-DAD-ESI-MS analysis supported this result, revealing the presence of 17-decarboxy betanin, 17-decarboxy neobetanin, and Cyclo-dopa5-O-βglucoside in DOS at 40 °C. This study highlights the potential future in the sustainable green extraction of betalain compounds with less heat degradation to offer a stable natural colorant. Full article
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<p>Desorption isotherms for <span class="html-italic">Opuntia stricta</span> fruit at 40 and 60 °C (<b>a</b>) Moisture ratio evolution versus drying time for <span class="html-italic">Opuntia stricta</span> fruit at 40 and 60 °C (<b>b</b>).</p>
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11 pages, 545 KiB  
Article
Improved Antioxidant Capacity by Block Cryoconcentration of Opuntia ficus-indica L. Mill (Green and Red) Juice
by Carlos Alberto Márquez-Montes, José Alberto Gallegos-Infante, Guillermo Rodrigo Petzold-Maldonado, Patricio Antonio Orellana-Palma, Rubén Francisco González-Laredo, Nuria Elizabeth Rocha-Guzmán and Martha Rocío Moreno-Jiménez
AppliedChem 2025, 5(1), 4; https://doi.org/10.3390/appliedchem5010004 - 19 Feb 2025
Viewed by 450
Abstract
The presence of bioactives in prickly pear has been documented, including flavonoids and betalains, which are compounds highly unstable to thermal processing. An alternative to the thermal processing of foods is the use of cryoconcentration. The objective of this work was to use [...] Read more.
The presence of bioactives in prickly pear has been documented, including flavonoids and betalains, which are compounds highly unstable to thermal processing. An alternative to the thermal processing of foods is the use of cryoconcentration. The objective of this work was to use cryoconcentration assisted by centrifugation to obtain prickly pear (Opuntia ficus-indica L. Mill) concentrate from two ecotypes (green and red) and evaluate their impact on the polyphenol profile and betalains. Prickly pear juice was obtained and cryoconcentrated. The process parameters of cryoconcentration were obtained. The highest solute yield (Y) was observed for red prickly pear juice (0.42 ± 0.03 kg solute × kg initial solute−1), but the efficiency (η) did not show differences between ecotypes (green 51.0 ± 7.0 vs. red 55.0 ± 7.0%), physicochemical parameters (pH, titratable acididty, °Bx), reducing sugars, or color. The highest total phenolic content (TPC) (1843 ± 153), total flavonoid content (TFC) (759 ± 17), betanin (801.6 ± 19), and indicaxanthin (453.7 ± 19) were observed in cryoconcentrated red prickly pear juice, while the antioxidant activity (ABTS, FRAP, and ORAC) was higher in cryoconcentrated green prickly pear juice (except ABTS). Betalains showed a high correlation with the ABTS antioxidant results, and the TPC showed a high correlation with the ORAC results. Cryoconcentration technology has a high potential to process prickly pear juice, preserving its bioactives. Full article
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<p>Images of prickly pear fruits (green and red) before washing and peeling as an example of the fruits used in the experiment.</p>
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37 pages, 2184 KiB  
Article
Exploring the Biochemical Profile of Beta vulgaris L.: A Comparative Study of Beetroots and Swiss Chard
by Daiana Almeida, Spyridon A. Petropoulos, Tayse F. F. da Silveira, Tânia C. S. P. Pires, Isabel C. F. R. Ferreira, Ângela Fernandes and Lillian Barros
Plants 2025, 14(4), 591; https://doi.org/10.3390/plants14040591 - 14 Feb 2025
Viewed by 425
Abstract
In this study, leaves and roots from three beetroot cultivars (cv. Albina Vereduna (white roots), cv. Burpee’s Golden (golden roots), and cv. Pablo F1 (red roots)), as well as Swiss chard leaves (also known as “rhubarb chard”, or Beta vulgaris subsp. cicla var. [...] Read more.
In this study, leaves and roots from three beetroot cultivars (cv. Albina Vereduna (white roots), cv. Burpee’s Golden (golden roots), and cv. Pablo F1 (red roots)), as well as Swiss chard leaves (also known as “rhubarb chard”, or Beta vulgaris subsp. cicla var. flavescens) were evaluated in terms of their chemical profile and bioactive properties. Roots were characterized by high carbohydrate content, which also contributed to greater energy values. In contrast, fibers were the predominant macronutrient in leaves, followed by carbohydrates. In both leaves and roots, the most abundant organic acids were quinic and oxalic, while the major free sugar was sucrose. The profile of fatty acid varied between the studied plant parts, with saturated fatty acids prevailing in root samples, while leaves exhibited higher levels of polyunsaturated fatty acids. Regarding phenolic composition, a total of 19 compounds were tentatively identified in leaves (including derivatives of vitexin, isorhamnetin, quercetin, and ferulic, sinapic, and p-coumaric acids), while the roots exhibited a less diverse composition, with a total of eight compounds identified (e.g., derivatives of ferulic, sinapic, p-coumaric and caffeic acids). A total of eight betalains were also identified, out of which seven were classified as betacyanins and one as betaxanthin. The leaves of Swiss chard presented compounds from both classes, while the roots and leaves of cv. Pablo F1 were characterized only by the presence of betacyanins, and those of cv. Burpee’s Golden only by betaxanthin. All samples exhibited relevant activity against Y. enterocolitica, L. monocytogenes, and S. aureus, although leaf samples demonstrated better antioxidant capacity. In conclusion, beetroot leaves outperformed their corresponding roots in terms of chemical composition, antioxidant, and antimicrobial activity, suggesting their high potential as nutrient-rich and functional ingredients in a diverse and well-balanced diet. Full article
(This article belongs to the Section Phytochemistry)
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<p>Fatty acid profile of Swiss chard leaves (<span class="html-italic">B. vulgaris</span> subsp. <span class="html-italic">cicla</span> var. <span class="html-italic">flavescens</span>) analyzed with GC-FID.</p>
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<p>Tocopherol profile of <span class="html-italic">B. vulgaris</span> cv. Burpee’s Golden leaves, analyzed with HPLC-FL. 1: α-tocopherol; 2: <span class="html-italic">β</span>-tocopherol; 3: <span class="html-italic">γ</span>-tocopherol; 4: tocol, IS.</p>
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<p>Betalain profile of Swiss chard leaves (<span class="html-italic">B. vulgaris</span> subsp. <span class="html-italic">cicla</span> var. <span class="html-italic">flavescens</span>) analyzed with LC-MS/MS. The peaks are identified in <a href="#plants-14-00591-t003" class="html-table">Table 3</a>.</p>
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<p>Phenolic profile of leaves from cv. Albina Vereduna analyzed with HPLC. The peaks are identified in <a href="#plants-14-00591-t004" class="html-table">Table 4</a>.</p>
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<p>Phenolic profile of roots from cv. Albina Vereduna analyzed with HPLC. The peaks are identified in <a href="#plants-14-00591-t005" class="html-table">Table 5</a>.</p>
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19 pages, 4526 KiB  
Article
Non-Targeted Metabolomics Reveals Tobacco Metabolites Associated with Roasted Tobacco Quality Under Organic Substitution
by Feng Tian, Li Zhang, Lina Niu, Xinglong Fan, Yongliang Han, Yuanshang Liu, Jiancai Qian, Shihang Huang, Xixian Ou and Qiang Li
Agronomy 2025, 15(2), 470; https://doi.org/10.3390/agronomy15020470 - 14 Feb 2025
Viewed by 274
Abstract
The present study investigated the effect of the replacement of chemical fertilizers by organic fertilizers on agronomic traits of tobacco at maturity as well as on the conventional chemical quality of post-roasted tobacco leaves. To better understand the relationship between tobacco metabolites and [...] Read more.
The present study investigated the effect of the replacement of chemical fertilizers by organic fertilizers on agronomic traits of tobacco at maturity as well as on the conventional chemical quality of post-roasted tobacco leaves. To better understand the relationship between tobacco metabolites and roasted tobacco under organic nitrogen replacement treatments, post-roasting tobacco leaves were analyzed by an untargeted metabolomics analytical approach to identify key metabolites applicable to predicting tobacco quality. Methods: Yunyun Tobacco 87 was adopted as the test material in a field plot experiment with five fertilization treatments: T1 (100% chemical nitrogen fertilizer), T2 (25% organic nitrogen fertilizer + 75% chemical nitrogen fertilizer), T3 (50% organic nitrogen fertilizer + 50% chemical nitrogen fertilizer), T4 (75% organic nitrogen fertilizer + 25% chemical nitrogen fertilizer), and T5 (100% organic nitrogen fertilizer). Additionally, the non-targeted metabolomics approach was employed for the in-depth analysis of metabolites in roasted tobacco leaves. Results: The targeted metabolomic analysis identified 991 metabolites in the positive ion pattern and 673 in the negative ion pattern across all treatments. Among these, certain pathways such as alanine, aspartate, and glutamate metabolism, D-amino acid metabolism, purine metabolism, tryptophan metabolism, and galactose metabolism were up-regulated, whereas other pathways such as starch and sucrose metabolism, betalain biosynthesis, and biosynthesis of unsaturated fatty acids were down-regulated and significantly enriched with differential metabolites. This study revealed the significant differences in the metabolite composition under different fertilization conditions, with a strong correlation between metabolites and tobacco quality indices. Organic fertilizers were observed to enhance tobacco quality by influencing tobacco metabolism, providing a scientific basis for optimizing fertilization strategies and improving tobacco quality. Full article
(This article belongs to the Special Issue Crop Genomics and Omics for Future Food Security)
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<p>Multivariate analysis of metabolites across different treatments. PCA result plots for metabolites in the negative ion mode (<b>a</b>) and positive ion mode (<b>b</b>), PLS-DA result plots for metabolites in the negative ion mode (<b>c</b>) and positive ion mode (<b>d</b>), and validation plots depicting the predictive performance of metabolites in the negative ion mode (<b>e</b>) and positive ion mode (<b>f</b>) assessed via 200 alignment tests.</p>
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<p>Metabolite difference analysis plots for 100% organic and 100% chemical fertilizer treatment groups. Volcano plots of negatively differential metabolites (<b>a</b>) and positively differential metabolites (<b>b</b>), and heat maps of the top 20 negatively differential metabolites (<b>c</b>), and the top 20 positively differential metabolites (<b>d</b>).</p>
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<p>Enrichment analysis plots of up-regulated metabolites from organic and chemical fertilizer treatments. Categorical bar graph of KEGG enrichment analysis of up-regulated metabolites (<b>a</b>), bubble graph of KEGG enrichment analysis of up-regulated metabolites (<b>b</b>), and string graph of enrichment analysis of up-regulated metabolites (<b>c</b>).</p>
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<p>Enrichment analysis plots of down-regulated metabolites from organic and chemical fertilizer treatments. Categorical bar graph of KEGG enrichment analysis of down-regulated metabolites (<b>a</b>), bubble graph of KEGG enrichment analysis of down-regulated metabolites (<b>b</b>), and string graph of enrichment analysis of down-regulated metabolites (<b>c</b>).</p>
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<p>Metabolite trend clustering analysis. Cluster 1–4 trend fitting plots (<b>a</b>–<b>d</b>) and heatmap of metabolite trend changes in each treatment (<b>e</b>).</p>
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<p>Mantel analysis plot of 21 differential metabolites with chemical indicators of roasted tobacco. “Mantel’s p” indicates the correlation between each differential metabolite and chemical indicator variables, “Mantel’s r” is used to measure the strength of correlation between differential metabolites and chemical indicators, and “Pearson’s r” indicates the linear correlation between each differential metabolite. *: indicates a significance level of <span class="html-italic">p</span> &lt; 0.05; **: indicates a significance level of <span class="html-italic">p</span> &lt; 0.01; ***: indicates a significance level of <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>Mantel analysis plot of 21 differential metabolites with yield, effective leaf number, and physical properties of roasted tobacco. *: indicates a significance level of <span class="html-italic">p</span> &lt; 0.05; **: indicates a significance level of <span class="html-italic">p</span> &lt; 0.01; ***: indicates a significance level of <span class="html-italic">p</span> &lt; 0.001.</p>
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24 pages, 15009 KiB  
Article
Differential Expression of Amaranth AtrDODA Gene Family Members in Betalain Synthesis and Functional Analysis of AtrDODA1-1 Promoter
by Huiying Xie, Jiajing Zeng, Wenli Feng, Wei Gao, Zhongxiong Lai and Shengcai Liu
Plants 2025, 14(3), 454; https://doi.org/10.3390/plants14030454 - 4 Feb 2025
Viewed by 549
Abstract
Betalains differ from anthocyanins, and they cannot coexist in the same plant under natural conditions. The L-DOPA 4,5-dioxygenase encoded by the DODA gene is a key step in the pathway of betalain biosynthesis in Caryophyllales plants. Amaranth is an important resource for the [...] Read more.
Betalains differ from anthocyanins, and they cannot coexist in the same plant under natural conditions. The L-DOPA 4,5-dioxygenase encoded by the DODA gene is a key step in the pathway of betalain biosynthesis in Caryophyllales plants. Amaranth is an important resource for the study and extraction of betalains. In order to clarify the function of AtrDODA family genes in betalain biosynthesis, we screened out three AtrDODA family gene members associated with betalains based on a genome database and RNA-seq databases of Amaranthus tricolor. Their characterization and expression pattern were further analyzed. The result of subcellular localization showed that all three AtrDODA members were located in the nucleus. Betacyanin and betaxanthin were promoted by paclobutrazol treatment in the leaves and stems of ‘Suxian No.1’ (red), while they were inhibited by gibberellin and darkness, which were consistent with the gene expression pattern of AtrDODAs. After heterologous transformation of the AtrDODA1-1 promoter into tobacco with GUS staining analysis, the promoter activity of AtrDODA1-1 of ‘Suxian No.1’ (red) amaranth was significantly higher than that of ‘Suxian No.2’ (green) amaranth. Furthermore, we analyzed the promoter activity of AtrDODA1-1 by GUS staining and qRT-PCR after sprayed exogenous MeJA and GA3 on the AtrDODA1-1 promoter transformed tobacco plants. The results showed that AtrDODA1-1 responded to plant hormones. This study could lay a foundation for revealing the biological functions of the amaranth DODA gene family, and provide new clues for the molecular mechanism of betalain synthesis. Full article
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<p>Structure of the <span class="html-italic">AtrDODA</span> gene, domain, motif, and chromosome localization in amaranthus: (<b>A</b>) Conserved domains of AtrDODAs; (<b>B</b>) Gene structure of <span class="html-italic">AtrDODAs</span>; (<b>C</b>) Chromosome localization of <span class="html-italic">AtrDODAs</span> genes; (<b>D</b>) Conserved motifs of AtrDODAs.</p>
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<p>Collinearity and phylogenetic tree of <span class="html-italic">AtrDODA</span> in amaranth: (<b>A</b>) Interspecific collinearity analysis of <span class="html-italic">DODA</span> between <span class="html-italic">Amaranthus tricolor</span> L., <span class="html-italic">Beta vulgaris</span>, and <span class="html-italic">Hylocereus undatus</span>. (<b>B</b>) Interspecific collinearity analysis of DODA between <span class="html-italic">Amaranthus tricolor</span>, <span class="html-italic">Portulaca oleracea</span>, and <span class="html-italic">Bougainvillea</span>. (<b>C</b>) Phylogenetic tree of <span class="html-italic">DODA</span> homologs between amaranth and other plants. The roman letters and different colors represents the four different groups.</p>
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<p>Multiple sequence alignment of DODA amino acids in amaranth and other species: Blue triangles represent Class I members of AtrDODA, and red triangles represent Class II members of AtrDODA. Asterisks indicate acidic residues that may be important for DOD activity/DODA recognition.</p>
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<p>Analysis of <span class="html-italic">cis</span>-acting elements in the promoter of <span class="html-italic">AtrDODA</span>. The numbers in the figure represent the counts of various <span class="html-italic">cis</span>-acting elements, with red indicating ‘Suxian No.1’ and green indicating ‘Suxian No.2’.</p>
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<p>Expression heatmap of <span class="html-italic">AtrDODA</span>: (<b>A</b>) Gene expression heatmap of <span class="html-italic">AtrDODA</span> in ‘Suxian No.1’ (red) under blue and white light quality, dark treatment, and plant hormone treatment; (<b>B</b>) Heatmap of <span class="html-italic">AtrDODA</span> gene expression in the red and green sections of the leaves of red amaranth.</p>
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<p>Expression patterns of <span class="html-italic">AtrDODA</span> genes in amaranth: (<b>A</b>) Phenotype images of different varieties and ‘Suxian No.1’ (red) amaranth under various treatments. (<b>a</b>): ‘Suxian No.2’ under white light treatment; (<b>b</b>): ‘Suxian No.1’ under white light treatment; (<b>c</b>): ‘Suxian No.1’ under dark treatment; (<b>d</b>): ‘Suxian No.1’ treated with 2 mg/L paclobutrazol; (<b>e</b>): ‘Suxian No.1’ treated with 1 mg/L gibberellin. (<b>B</b>) <span class="html-italic">AtrDODA1-1</span>, (<b>C</b>) <span class="html-italic">AtrDODA2-1</span>, and (<b>D</b>) <span class="html-italic">AtrDODA1-2</span> expression patterns in ‘Suxian No.1’ and ‘Suxian No.2’ under white light treatment. (<b>E</b>) <span class="html-italic">AtrDODA1-1</span>, (<b>F</b>) <span class="html-italic">AtrDODA2-1</span>, and (<b>G</b>) <span class="html-italic">AtrDODA1-2</span> expression patterns in ‘Suxian No.1’ under white light, gibberellin, paclobutrazol, and dark treatments. (<b>H</b>) Expression patterns of <span class="html-italic">AtrDODAs</span> genes under different concentrations of salt treatment. The scale bar in A represents 1 cm, and different lowercase letters in (<b>B</b>–<b>H</b>) represent significant differences in gene expression (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Phenotypes and betacyanin content in amaranth: (<b>A</b>) Betacyanin samples from ‘Suxian No.2’ (green) amaranth under white light treatment; (<b>B</b>) Betacyanin samples from ‘Suxian No.1’ (red) amaranth under white light treatment, (<b>C</b>) under gibberellin treatment, (<b>D</b>) under paclobutrazol treatment, (<b>E</b>) and under dark treatment; (<b>F</b>) Betacyanin content in roots, stems, and leaves of ‘Suxian No.1’ (red) and ‘Suxian No.2’ (green) amaranth; (<b>G</b>) Betaxanthin content in roots, stems, and leaves of ‘Suxian No.1’ (red) and ‘Suxian No.2’ (green) amaranth; (<b>H</b>) Betacyanin content and (<b>I</b>) betaxanthin content in ‘Suxian No.1’ (red) amaranth under different light qualities, gibberellin, and paclobutrazol treatments. The scale bar in (<b>A</b>–<b>E</b>) is 1 cm, and different lowercase letters in (<b>F</b>–<b>I</b>) represent significant differences in betacyanin and betaxanthin content (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Subcellular localization of AtrDODAs. DAPI panel represents green fluorescence co-localized with DAPI staining; GFP panel represents GFP fluorescence; Bright panel represents bright field photograph; Merge panel represents bright field overlay of GFP fluorescence.</p>
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<p>Analysis of promoter activity of AtrDODA1-1 in red and green amaranth: (<b>A</b>) GUS staining of wild-type control tobacco; (<b>B</b>) GUS staining of tobacco with empty vector pCAMBIA1301; (<b>C</b>) GUS staining of tobacco with the promoter of red AtrDODA1-1; (<b>D</b>) GUS staining of tobacco with the promoter of green AtrDODA1-1; (<b>E</b>) β-Glucuronidase (GUS) enzyme activity; (<b>F</b>) Expression of the GUS gene. The different lowercase letters in (<b>E</b>,<b>F</b>) represent significant differences (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Analysis of the <span class="html-italic">AtrDODA1-1</span> promoters in red and green amaranth in response to exogenous hormone treatments: (<b>A</b>) Construction of <span class="html-italic">GUS</span> expression vectors for the <span class="html-italic">AtrDODA1-1</span> promoters of red and green amaranth; (<b>B</b>) GUS staining of tobacco with the red <span class="html-italic">AtrDODA1-1</span> promoter under MeJA and GA treatments; (<b>C</b>) GUS staining of tobacco with the <span class="html-italic">green AtrDODA1-1</span> promoter under MeJA and GA treatments; (<b>D</b>) β-Glucuronidase (GUS) enzyme activity and <span class="html-italic">GUS</span> gene expression of the <span class="html-italic">red AtrDODA1-1</span> promoter under MeJA and GA treatments; (<b>E</b>) β-Glucuronidase (GUS) enzyme activity and <span class="html-italic">GUS</span> gene expression of the <span class="html-italic">green AtrDODA1-1 promoter</span> under MeJA and GA treatments. In (<b>D</b>,<b>E</b>), different lowercase letters represent significant differences in gene expression (<span class="html-italic">p</span> &lt; 0.05).</p>
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22 pages, 3975 KiB  
Article
Feasibility and Tolerability of Daily Microgreen Consumption in Healthy Middle-Aged/Older Adults: A Randomized, Open-Label, Controlled Crossover Trial
by Sylvia Y. Lee, Kiri A. Michell, Michelle M. Butler, Brayden T. Smith, Emily K. Woolf, Sydney C. Holmes, Lauren E. Grabos, Allegra R. Vazquez, Hanan Isweiri, Marisa Bunning, Mark E. Uchanski, Sangeeta Rao, Steven E. Newman, Tiffany L. Weir and Sarah A. Johnson
Nutrients 2025, 17(3), 467; https://doi.org/10.3390/nu17030467 - 28 Jan 2025
Viewed by 820
Abstract
Background/Objectives: Microgreens are rich in nutrients and phytochemicals that can support healthy aging, including attenuation of cardiovascular disease risk. The nutrient and phytochemical contents of red beet (i.e., bull’s blood’ beet, Beta vulgaris) and red cabbage (Brassica oleracea var capitate) microgreens, [...] Read more.
Background/Objectives: Microgreens are rich in nutrients and phytochemicals that can support healthy aging, including attenuation of cardiovascular disease risk. The nutrient and phytochemical contents of red beet (i.e., bull’s blood’ beet, Beta vulgaris) and red cabbage (Brassica oleracea var capitate) microgreens, as well as existing preclinical evidence suggest their cardioprotective effects, but the feasibility, gastrointestinal tolerability, and human health effects of daily microgreen consumption are unknown. This study aimed to evaluate the feasibility and gastrointestinal tolerability of 2 weeks of daily microgreen consumption in healthy middle-aged/older (MA/O) adults. A secondary aim was to characterize potential health effects. Methods: Healthy MA/O adults (initial n = 26) were randomized to consume either 2 cups of ‘bull’s blood’ beet or red cabbage microgreens daily for 2 weeks in a crossover design, with each treatment period separated by 2 weeks. Feasibility was determined through participant retention and intervention compliance (i.e., total doses consumed divided by 14 days), while gastrointestinal tolerability was determined by a gastrointestinal health questionnaire and bowel movement log. Impacts of microgreen consumption on brachial and aortic hemodynamic parameters, and gut microbiota composition were evaluated. Results: Daily consumption for 2 weeks of ‘bull’s blood’ beet and red cabbage microgreens was found to be feasible as indicated by high participant retention (final n = 24) and overall treatment compliance of 95.6%. Gastrointestinal symptom severity was not impacted overall, though an improvement in gastrointestinal inflammation-associated symptom severity scores following the red cabbage microgreen intervention (p = 0.047) was observed. There were no changes in bowel movement quality, hemodynamic parameters, or on alpha or beta diversity of the gut microbiota. Conclusions: Daily consumption of ‘bull’s blood’ beet and red cabbage microgreens is feasible and tolerable in healthy MA/O adults. Future studies designed to evaluate their health impacts are needed. Full article
(This article belongs to the Section Phytochemicals and Human Health)
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<p>CONSORT flow diagram of participants through a randomized, open-label, 2-period crossover trial where they consumed 2 cups/day of ‘bull’s blood’ beet microgreens or red cabbage microgreens for a 2-week period.</p>
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<p>Schematic of clinical trial design and data collection in a randomized, open-label, 2-period crossover trial where they consumed 2 cups/day of ‘bull’s blood’ beet microgreens or red cabbage microgreens for a 2-week period. Procedures completed at each visit are listed with an X. Abbreviations: ASA24, automated self-administered 24-h dietary recall; PWA, pulse wave analysis.</p>
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<p>Changes in gastrointestinal symptom severity priority level following daily microgreen consumption as determined by a gastrointestinal health questionnaire in a randomized, open-label, 2-period crossover trial where they consumed 2 cups/day of ‘bull’s blood’ beet microgreens or red cabbage microgreens for a 2-week period. (<b>A</b>) total gastrointestinal symptom severity scores, (<b>B</b>) gastric function symptom severity scores, (<b>C</b>) gastrointestinal inflammation symptom severity scores, (<b>D</b>) small intestine and pancreas function symptom severity scores, and (<b>E</b>) colon function symptom severity scores. Data are mean ± SEM. * <span class="html-italic">p</span> &lt; 0.05. ‘Bull’s Blood’ Beet n = 25; Red Cabbage n = 24. Axes are colored by symptom priority category (low-high). Abbreviations: GI, gastrointestinal; ns, not significant; SI, small intestine.</p>
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<p>Gastric function symptom severity priority level before and after daily microgreen consumption as determined by a gastrointestinal health questionnaire in a randomized, open-label, 2-period crossover trial where they consumed 2 cups/day of ‘bull’s blood’ beet microgreens or red cabbage microgreens for a 2-week period. Change in priority level for (<b>A</b>) ‘bull’s blood’ beet microgreens and number of participants in each priority level at (<b>B</b>) baseline and (<b>C</b>) 2 weeks following ‘bull’s blood’ beet (n = 25); and change in priority level for (<b>D</b>) red cabbage microgreens and number of participants in each priority level at (<b>E</b>) baseline and (<b>F</b>) 2 weeks following red cabbage (n = 24) microgreens consumption.</p>
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<p>Gastrointestinal inflammation symptom severity priority level before and after daily microgreen consumption as determined by a gastrointestinal health questionnaire in a randomized, open-label, 2-period crossover trial where they consumed 2 cups/day of ‘bull’s blood’ beet microgreens or red cabbage microgreens for a 2-week period. Change in priority level for (<b>A</b>) ‘bull’s blood’ beet microgreens and number of participants in each priority level at (<b>B</b>) baseline and (<b>C</b>) 2 weeks following ‘bull’s blood’ beet (n = 25); and change in priority level for (<b>D</b>) red cabbage microgreens and number of participants in each priority level at (<b>E</b>) baseline and (<b>F</b>) 2 weeks following red cabbage (n = 24) microgreens consumption.</p>
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<p>Small intestine and pancreas function symptom severity priority level before and after daily microgreen consumption as determined by a gastrointestinal health questionnaire in a randomized, open-label, 2-period crossover trial where they consumed 2 cups/day of ‘bull’s blood’ beet microgreens or red cabbage microgreens for a 2-week period. Change in priority level for (<b>A</b>) ‘bull’s blood’ beet microgreens and number of participants in each priority level at (<b>B</b>) baseline and (<b>C</b>) 2 weeks following ‘bull’s blood’ beet (n = 25); and change in priority level for (<b>D</b>) red cabbage microgreens and number of participants in each priority level at (<b>E</b>) baseline and (<b>F</b>) 2 weeks following red cabbage (n = 24) microgreens consumption.</p>
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<p>Colon function symptom severity priority level before and after daily microgreen consumption as determined by a gastrointestinal health questionnaire in a randomized, open-label, 2-period crossover trial where they consumed 2 cups/day of ‘bull’s blood’ beet microgreens or red cabbage microgreens for a 2-week period. Change in priority level for (<b>A</b>) ‘bull’s blood’ beet microgreens and number of participants in each priority level at (<b>B</b>) baseline and (<b>C</b>) 2 weeks following ‘bull’s blood’ beet (n = 25); and change in priority level for (<b>D</b>) red cabbage microgreens and number of participants in each priority level at (<b>E</b>) baseline and (<b>F</b>) 2 weeks following red cabbage (n = 24) microgreens consumption.</p>
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<p>Average bowel movement frequency and type was evaluated through a bowel movement log (i.e., Bristol stool scale) in a randomized, open-label, 2-period crossover trial where they consumed 2 cups/day of ‘bull’s blood’ beet microgreens or red cabbage microgreens for a 2-week period. (<b>A</b>) Number of bowel movements per day and (<b>B</b>) Bristol stool scale bowel movement type comparisons between the first vs. second week of microgreens intervention, (<b>C</b>) number of bowel movements per day and (<b>D</b>) Bristol stool scale bowel movement type comparisons between the first two days vs. the second week of intervention, (<b>E</b>) number of bowel movements per day and (<b>F</b>) Bristol stool scale bowel movement type comparisons between day 1 vs. day 2 of intervention, (<b>G</b>) number of bowel movements per day and (<b>H</b>) Bristol stool scale BM type comparisons between the day 1 vs. day 3 of the intervention. Data are mean ± SEM. ‘Bull’s Blood’ Beet n = 25; Red Cabbage n = 24. Participants with identical bowel movement data overlap and appear as one point. Abbreviations: BM, bowel movement.</p>
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<p>Gut microbiota parameters at baseline and 2-weeks following daily consumption of 2 cups/day of ‘bull’s blood’ beet microgreens or red cabbage microgreens in a randomized, open-label, 2-period crossover trial. (<b>A</b>) Relative abundance of taxa at the phylum level, (<b>B</b>) alpha diversity of the gut microbiota at the genus level measured by Shannon index, and (<b>C</b>) beta diversity of the gut microbiota PCoA plots at the genus level measured by Bray-Curtis. ‘Bull’s blood’ beet n = 24; red cabbage n = 23.</p>
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39 pages, 3037 KiB  
Review
Red Beetroot and Its By-Products: A Comprehensive Review of Phytochemicals, Extraction Methods, Health Benefits, and Applications
by Florina Stoica, Gabriela Râpeanu, Roxana Nicoleta Rațu, Nicoleta Stănciuc, Constantin Croitoru, Denis Țopa and Gerard Jităreanu
Agriculture 2025, 15(3), 270; https://doi.org/10.3390/agriculture15030270 - 26 Jan 2025
Viewed by 1082
Abstract
Beetroot (Beta vulgaris), a root vegetable known for its vivid natural color and nutritional profile, is a source of a wide range of bioactive compounds, including betalains, phenolics, vitamins, and antioxidants. These bioactive compounds are associated with many health-promoting properties, including [...] Read more.
Beetroot (Beta vulgaris), a root vegetable known for its vivid natural color and nutritional profile, is a source of a wide range of bioactive compounds, including betalains, phenolics, vitamins, and antioxidants. These bioactive compounds are associated with many health-promoting properties, including antihypertensive, antioxidant, anti-inflammatory, and anticancer effects. The beetroot processing industry produces substantial by-products abundant in phytochemicals and betalains, presenting valuable opportunities for utilization. Therefore, it can replace synthetic additives and enhance the nutritional value of foods. By reducing waste and supporting a circular economy, beetroot by-products improve resource efficiency, cut production costs, and lessen the food industry’s environmental impact. Beetroot and its by-products are rich in phytochemicals that provide various wellness advantages. They support cardiovascular health, inhibit microbe-induced food spoiling, aid liver function, and reduce inflammation and oxidative stress. This paper presents a detailed review of current knowledge on beetroot and its by-products, focusing on their biochemical components, extraction and stabilization techniques, health benefits, and potential applications in the food industry. It underscores the versatility and importance of red beetroot and its derivatives, advocating for further research into optimized processing methods and innovative uses to enhance their industrial and nutritional value. By providing valuable insights, this review aims to inspire food scientists, nutritionists, and the agricultural sector to integrate beetroot and its by-products into more sustainable and health-oriented food systems. Full article
(This article belongs to the Section Agricultural Product Quality and Safety)
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<p><span class="html-italic">Beta vulgaris</span> L. around the world [<a href="#B12-agriculture-15-00270" class="html-bibr">12</a>]. The different colors express overlapping classic hexagons (56,848 georeferenced records) and mean the occurrences of subspecies, species, and varieties of beetroot species in each area of the globe.</p>
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<p>Biochemical constituents of beetroot by-products and their health-related effects.</p>
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<p>Examples of structures of various important bioactive compounds in beetroot by-products. (<b>a</b>) β-carotene, (<b>b</b>) Betanin, (<b>c</b>) Vulgaxanthin-I, (<b>d</b>) Ferulic acid, (<b>e</b>) Kaempferol, (<b>f</b>) Rutin [<a href="#B51-agriculture-15-00270" class="html-bibr">51</a>].</p>
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<p>Valorization of beetroot by-products and potential products.</p>
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<p>Challenges and future insights of valorization beetroot by-products.</p>
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20 pages, 18177 KiB  
Article
Identification of R2R3-MYB Transcription Factor Family Based on Amaranthus tricolor Genome and AtrMYB72 Promoting Betalain Biosynthesis by Directly Activating AtrCYP76AD1 Expression
by Yuwei Xue, Kexuan Li, Wenli Feng, Zhongxiong Lai and Shengcai Liu
Plants 2025, 14(3), 324; https://doi.org/10.3390/plants14030324 - 22 Jan 2025
Viewed by 637
Abstract
MYB (myeloblastosis) is one of the most abundant transcription factors in plants which regulates various biological processes. The molecular characteristics and function of R2R3-MYB transcription factors in amaranth remain unclear. In this study, 73 R2R3-MYB members were identified from the amaranth genome database [...] Read more.
MYB (myeloblastosis) is one of the most abundant transcription factors in plants which regulates various biological processes. The molecular characteristics and function of R2R3-MYB transcription factors in amaranth remain unclear. In this study, 73 R2R3-MYB members were identified from the amaranth genome database and we further analyzed their chromosome position, conserved motifs, physiological and biochemical features, collinearity relationships, gene structure, phylogeny and cis-acting element. Based on the phylogenetic and expression pattern analysis, 14 candidate R2R3-MYB genes might be involved in the betalain synthesis. Amongst the 14 candidate R2R3-MYB genes, the expression level of AtrMYB72 was higher in ‘Suxian No.1’ than ‘Suxian No.2’, and also higher in the red section than in the green section of the same leaf in Amaranthus. The overexpression vector pCambia1301-AtrMYB72-GUS and VIGS (virus-induced gene silencing) vector pTRV2- AtrMYB72 were transferred into leaves of ‘Suxian No.1’ via an Agrobacterium-mediated method. The results showed that AtrMYB72 overexpression could promote betalain synthesis. A yeast one-hybrid assay and dual luciferase reporter gene assay demonstrated that AtrMYB72 could bind to the AtrCYP76AD1 promoter to promote betalain synthesis. These results indicated that AtrMYB72 promoted betalain biosynthesis in amaranth by activating the AtrCYP76AD1 transcription. Our results could provide new insights into the betalain biosynthesis in amaranth. Full article
(This article belongs to the Special Issue Bioinformatics and Functional Genomics in Modern Plant Science)
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<p>Distribution of <span class="html-italic">Amaranthus tricolor R2R3-MYB</span> (<span class="html-italic">AtrMYB</span>) genes among 17 chromosomes. Gene positions and the size of each chromosome can be estimated using the scale on the right of the figure; the scale indicates 10 megabases (Mb).</p>
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<p>Comparison of DNA-binding domains of R2R3-MYB transcription factor in <span class="html-italic">Amaranthus tricolor</span>, <span class="html-italic">Hylocereus undatus</span>, <span class="html-italic">Beta vulgaris</span> and <span class="html-italic">Arabidopsis thaliana</span>. Sequence logos of the R2 and R3 repeats are based on conserved alignments from <span class="html-italic">Amaranthus tricolor</span> (<b>A</b>), <span class="html-italic">Hylocereus undatus</span> (<b>B</b>), <span class="html-italic">Beta vulgaris</span> (<b>C</b>) and <span class="html-italic">Arabidopsis thaliana</span> (<b>D</b>). The overall height of each stack indicates the conservation of the sequence at the position, whereas the height of letters within each stack represents the relative frequency of the corresponding amino acid. Highly conserved tryptophan (W) and phenylalanine (F) residues are indicated by yellow asterisks. The positions with different patterns between <span class="html-italic">Amaranthus tricolor</span>, <span class="html-italic">Hylocereus undatus</span>, <span class="html-italic">Beta vulgaris</span> and <span class="html-italic">Arabidopsis thaliana</span> are indicated by arrows. The positions with different patterns between <span class="html-italic">Amaranthus tricolor</span>, <span class="html-italic">Hylocereus undatus</span>, <span class="html-italic">Beta vulgaris</span> and <span class="html-italic">Arabidopsis thaliana</span> are indicated by red triangle.</p>
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<p>Collinearity analyses of <span class="html-italic">R2R3-MYB</span> genes. (<b>A</b>) Segmental duplication events of <span class="html-italic">R2R3-MYB</span> genes in amaranth. (<b>B</b>) Duplication events of <span class="html-italic">R2R3-MYB</span> genes between amaranth and pitaya. (<b>C</b>) Duplication events of <span class="html-italic">R2R3-MYB</span> genes between amaranth and beet. (<b>D</b>) Duplication events of <span class="html-italic">R2R3-MYB</span> genes between amaranth and <span class="html-italic">Arabidopsis thaliana</span>. Purple lines indicate duplication events of <span class="html-italic">R2R3-MYB</span> genes. Gray lines represent all synteny blocks in genomes.</p>
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<p>Phylogenetic relationships of R2R3-MYBs. <span class="html-italic">Arabidopsis thaliana</span>, amaranth, pitaya and beet R2R3-MYBs were used for the phylogenetic tree construction using the ML method. Red stars represent the R2R3-MYBs of amaranth, blue circles represent the R2R3-MYBs of pitaya, green circles symbolize the R2R3-MYBs of <span class="html-italic">Arabidopsis thaliana</span> and yellow circles represent the R2R3-MYBs of beet.</p>
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<p>AtrR2R3-MYBs phylogenetic relationship (<b>A</b>), conserved motifs (<b>B</b>), and conserved domains (<b>C</b>). Orange circles indicate the bootstrap value range from 81 to 100 in the tree, green is from 60 to 80, and blue is from 0 to 59.</p>
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<p>The regulatory element of R2R3-MYB gene promoters in amaranth.</p>
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<p>Expression patterns of the AtrMYBs.</p>
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<p>Quantitative analysis of selected <span class="html-italic">AtrR2R3-MYBs</span>. (<b>A</b>) Quantitative expression analysis in the leaves of ‘Suxian No.1’ and ‘Suxian No.2’; (<b>B</b>) Quantitative expression in the stems of ‘Suxian No.1’ and ‘Suxian No.2’; (<b>C</b>) Quantitative expression in the different sections of <span class="html-italic">Amaranthus</span> leaves. * indicates significant differences at <span class="html-italic">p</span> &lt; 0.05, ** indicates significant differences at <span class="html-italic">p</span> &lt; 0.01, *** indicates significant differences at <span class="html-italic">p</span> &lt; 0.001, and **** indicates significant differences at <span class="html-italic">p</span> &lt; 0.0001.</p>
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<p><span class="html-italic">Agrobacterium</span>-mediated transient transformation into the amaranth leaves revealing that the overexpression of <span class="html-italic">AtrMYB72</span> promotes the betalain synthesis in amaranth. (<b>A</b>) Plant leaves after transient transformation for 7 days. (<b>B</b>) Plants after transient transformation for 7 days. (<b>C</b>) Relative expression of betalain synthesis-related genes in leaves of plants with different transient transformations. (<b>D</b>) Betalain contents in the leaves with different transient transformation plants. (a, b and c indicate significant differences at <span class="html-italic">p</span> &lt; 0.05; Bars: 1 cm).</p>
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<p>Silencing of <span class="html-italic">AtrMYB72</span> inhibited betalain synthesis. (<b>A</b>) control leaves (<b>left</b>), VIGS-empty leaves (<b>middle</b>) and VIGS-MYB72 leaves (<b>right</b>). (<b>B</b>) control plant (<b>left</b>), VIGS-empty plant (<b>middle</b>) and VIGS-MYB72 plant (<b>right</b>). (<b>C</b>) Relative expression levels of key genes involved in betalain synthesis in transgenic plants with gene silencing. (<b>D</b>) Betalain contents in leaves with gene silencing plants. Three biological replicates were performed for each sample (a, b and c indicate significant differences at <span class="html-italic">p</span> &lt; 0.01; Bar = 1 cm).</p>
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<p>Y1H assay of <span class="html-italic">AtrMYB72</span> with <span class="html-italic">AtrCYP76AD1</span> promoter. The promoter of <span class="html-italic">AtrCYP76AD1</span> was constructed in the pHis2 vector, and the ORF of <span class="html-italic">AtrMYB72</span> was constructed in the pGADT7 vector. Yeast cells were cultured on an SD/-Leu-Trp-His medium supplemented with 100 nm of 3-AT.</p>
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<p><span class="html-italic">AtrMYB72</span> promoted the <span class="html-italic">AtrCYP76AD1</span> transcription in <span class="html-italic">Nicotiana benthamiana</span> leaves. ** indicates significant differences at <span class="html-italic">p</span> &lt; 0.01.</p>
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<p>A hypothetical model of <span class="html-italic">AtrMYB72</span> gene regulated <span class="html-italic">AtrCYP76AD1</span> involved in betalain biosynthesis in amaranth. <span class="html-italic">AtrMYB72</span> transcript factor activated <span class="html-italic">AtrCYP76AD1</span> transcription by binding the MBS elements of the <span class="html-italic">AtrCYP76AD1</span> promoter.</p>
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<p>Plant phenotype of amaranth. (<b>A</b>) represents ‘Suxian No.1’ and ‘Suxian No.2’. (<b>B</b>) represents different parts in full-red amaranth leaves. (<b>C</b>) represents ‘Suxian No.1’ in (2000 lux, 16 h light/8 h dark, temperature 26 ± 1 °C). (<b>D</b>) represents ‘Suxian No.1’ in (8000 lux, 16 h light/8 h dark, temperature 26 ± 1 °C).</p>
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19 pages, 1531 KiB  
Article
A Sustainable Approach: Repurposing Red Beetroot Peels for Innovative Meringue Products
by Oana Emilia Constantin, Florina Stoica, Silvia Lazăr (Mistrianu), Doina Georgeta Andronoiu, Mihaela Turturică, Nicoleta Stănciuc, Roxana Nicoleta Rațu, Constantin Croitoru and Gabriela Râpeanu
Foods 2025, 14(2), 317; https://doi.org/10.3390/foods14020317 - 18 Jan 2025
Cited by 1 | Viewed by 861
Abstract
With the increasing global demand for sustainable and eco-friendly food items, it is imperative to investigate alternate sources of natural pigments. The red beetroot (Beta vulgaris L.) is a traditional food in many countries and a rich bioactive compound known for its [...] Read more.
With the increasing global demand for sustainable and eco-friendly food items, it is imperative to investigate alternate sources of natural pigments. The red beetroot (Beta vulgaris L.) is a traditional food in many countries and a rich bioactive compound known for its beneficial properties. Beetroot peel, a by-product of beetroot food processing, is often discarded, contributing to environmental damage. This research explores the potential of beetroot peel (BP) powder as a natural pigment in food products and its functional benefits. The study focuses on incorporating BP powder into meringues, aiming to create a value-added product with enhanced properties, particularly antioxidant activity. Various amounts of BP powder (4–10%) were added to meringue formulations, and the effects on the resulting meringues’ physicochemical properties, sensory qualities, and phytochemical profiles were assessed during 21 days of storage. The research revealed that BP powder, besides its function as a natural colorant and the pleasing pink hue it imparts to meringues, also enhances antioxidant activity due to its high phenolic concentration. BP powder was also incorporated to enhance the meringues’ overall sensory characteristics, improving their flavor and texture. The research findings indicate that BP has the potential to be used as a natural food ingredient to promote human health, resource-use efficiency, and a circular economy. Full article
(This article belongs to the Special Issue Comprehensive Utilization of By-Products in Food Industry)
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<p>HPLC chromatograms of betalains quantified in the extract of BP.</p>
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<p>Comparative diagram of the sensory attributes specific to meringues: C-meringues without adding BP powder; M1, M2, and M3—meringues with 4, 7, and 10% powder of BP.</p>
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<p>Meringues with different percentages of BP powder: C(control)-meringues without added BP powder, M1, M2, and M3—meringues with 4, 7, and 10% added BP powder.</p>
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<p>Illustration of correlations among sensory characteristics through Principal Component Analysis (PCA).</p>
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17 pages, 2757 KiB  
Article
Betalain–Chickpea Protein Particles Produced by Freeze Drying and Spray Drying: Physicochemical Aspects, Storage Stability, and In Vitro Digestion
by Mary H. Grace, Roberta Targino Hoskin, Malak Alghamdi, Mary Ann Lila and Vesela I. Chalova
Foods 2025, 14(2), 281; https://doi.org/10.3390/foods14020281 - 16 Jan 2025
Viewed by 909
Abstract
Beetroots are one of the primary sources of betalains, nitrogenous pigments with anti-inflammatory and antioxidant properties. However, due to their chemical instability, betalains have limited use in food applications. This work investigated whether betalains encapsulated in chickpea protein could be stabilized and delivered [...] Read more.
Beetroots are one of the primary sources of betalains, nitrogenous pigments with anti-inflammatory and antioxidant properties. However, due to their chemical instability, betalains have limited use in food applications. This work investigated whether betalains encapsulated in chickpea protein could be stabilized and delivered in a shelf-stable format. Freeze-dried (CB-FD) and spray-dried (CB-SD) protein–betalain particles encapsulated in chickpea protein isolate (6% w/v) were prepared. The encapsulation method affected particles’ morphology, water activity, hygroscopicity, solubility, and color. Particles captured total betalains of 9.30 ± 0.61 and 4.40 ± 0.92 mg/g for CB-SD and CB-FD, respectively. LC-MS identified 12 betacyanins and 6 betaxanthins. The stability of betalains revealed that encapsulation efficiently preserved betalain integrity of over 6 weeks of storage at 4, 22, and 40 °C compared to dry beetroot extract. CB-SD particles were stable with no significant changes, while CB-FD showed slight degradation after 4 weeks due to increased Aw. Antioxidant activity correlated well with betalain concentration. In vitro digestion resulted in only 25% bioaccessibility of betacyanins, while betaxanthins were more stable with 100% recovery. Encapsulation with chickpea protein isolate is an efficient and straightforward strategy for expanding and diversifying applications of phytochemical-rich beetroot extracts for the food industry. Full article
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<p>Concentrated beetroot extract (<b>A</b>), chickpea protein isolate (<b>B</b>), freeze-dried chickpea protein–betalain particles (<b>C</b>) and spray-dried chickpea protein–betalain particles (<b>D</b>).</p>
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<p>Scanning electron micrographs of CB-SD: spray-dried chickpea protein–betalain particles (<b>A</b>–<b>C</b>) and CB-SD: freeze-dried chickpea protein–betalain particles (<b>D</b>–<b>F</b>). Images (<b>A</b>,<b>D</b>): magnification 250×; images (<b>B</b>,<b>E</b>): magnification 500×; images (<b>C</b>,<b>E</b>): magnification 1000×.</p>
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<p>HPLC profiles of betalains recorded at 480 nm showing the major betxanthin (Vulgaxanthin I) and major betacyanins (Betanin, Isobetanin and Neobetanin) (<b>A</b>) and betacyanins (Betanin, Isobetanin) recorded at 532 nm (<b>B</b>).</p>
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<p>A<span class="html-italic"><sub>w</sub></span> (<b>A</b>), ΔE total color difference (<b>B</b>), L* (<b>C</b>), a* (<b>D</b>), and b* (<b>E</b>) results of protein–betalain particles during 28-day storage at different temperatures. Legend: CB-FD 4C: freeze-dried chickpea protein–betalain particles stored at 4 °C; CB-FD 22C: freeze-dried chickpea protein–betalain particles stored at 22 °C; CB-FD 40 C: freeze-dried chickpea protein–betalain particles stored at 40 °C; CB-SD 4C: spray-dried chickpea protein–betalain particles stored at 4 °C; CB-SD 22C: spray-dried chickpea protein–betalain particles stored at 22 °C; CB-SD 40 C: spray-dried chickpea protein–betalain particles stored at 40 °C. Asterisks indicate significant statistical differences by ANOVA analysis and Dunnett’s test between samples analyzed after 2 or 4 weeks of storage and right after production (week 0). *: <span class="html-italic">p</span> ≤ 0.05; **: <span class="html-italic">p</span> ≤0.01; ***: <span class="html-italic">p</span> ≤ 0.001; ****: <span class="html-italic">p</span> ≤ 0.0001. For clarity, some asterisks are shown on the side of the experimental point.</p>
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<p>Retention of betacyanins (<b>A</b>), betaxanthins (<b>B</b>), and DPPH antioxidant capacity (<b>C</b>) of beetroot extract (BEx), freeze-dried chickpea protein–betalain particles (BC-FD) and spray-dried chickpea protein–betalain particles (CB-SD) during six weeks of storage at 5 °C, 22 °C, and 40 °C. (<b>D</b>) Bioaccessibility index of betacyanins and betaxanthins after intestinal phase digestion. According to ANOVA analysis and Dunnett’s test, asterisks indicate significant differences between samples analyzed after 2, 4, or 6 weeks of storage and right after production (week 0). *: <span class="html-italic">p</span> ≤ 0.05; **: <span class="html-italic">p</span> ≤0.01; ***: <span class="html-italic">p</span> ≤ 0.001; ****: <span class="html-italic">p</span> ≤ 0.0001.</p>
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19 pages, 7230 KiB  
Article
Conservation Analysis and Colorimetric Characterization of Betalain Extracts from the Peel of Red Beetroot, Golden Beetroot, and Prickly Pear Applied to Cottage Cheese
by Elizabeth López-Solórzano, Claudia Muro, Yolanda Alvarado Perez, Andrea Y. Guadarrama-Lezama, Elsa Gutiérrez-Cortez and Juan Manuel Urrieta
Foods 2025, 14(2), 228; https://doi.org/10.3390/foods14020228 - 13 Jan 2025
Viewed by 745
Abstract
The maintenance of betalains and the color of extracts from the peel of red beetroot (RBAC), golden beetroot (YBAC), and purple prickly pear (PBAC) were evaluated, describing the capacity of their use as natural pigments and in the formulation of attractive and functional [...] Read more.
The maintenance of betalains and the color of extracts from the peel of red beetroot (RBAC), golden beetroot (YBAC), and purple prickly pear (PBAC) were evaluated, describing the capacity of their use as natural pigments and in the formulation of attractive and functional foods. Betalain extracts were prepared as juices from frozen and dehydrated peel, adding organic acids and concentrating for water reduction. Extracts were evaluated and applied on cottage cheese, measuring the capacity of betalains retention and pigmentation, during 10 days of storage of closed and opened products. Extracts of RBAC showed the highest betacyanin concentration, followed by YBAC with betaxanthins and PBAC with less betacyanin content. The pH stability for the extracts was pH4–7; RBAC and PBAC were stables at <90 °C, whereas YBAC exposed >125 °C. Extracts were constant during 10 days under oxygen and light exposure; however, YBAC exhibited low resistance in this environment. With cottage cheese, extracts exposed no changes in betalains and color on closed products (10 days of storage at 4 °C). In opened products, PBAC maintained the maximum betalains and color at 90%, PBAC at 75%, and YBAC at 60%. The obtained data contributed to use of agro-industrial residues, betalain extraction and conservation, and their potential use in food coloration and stabilization. Full article
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<p>Uv-vis spectra and images of betalain extracts from red beetroot, golden beetroot, and purple prickly pear from samples of RBAC, YBAC and PBAC.</p>
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<p>The color characteristics Chroma* and Hue° of the betalain extracts of RB, RBA, RBC RBAC, YB, YBA, YBC, YBAC, PB, PBA, PBC, and PBAC.</p>
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<p>Kinetics of betalains maintenance in extracts RBAC, YBAC, and PBAC.</p>
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<p>Luminosity L* and color difference ΔE from samples of cottage cheese with betalain extracts; conforming samples CCH + PBAC, CCH + RBAC and CCH-YBAC for closed products (above) and opened products (below).</p>
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22 pages, 6199 KiB  
Article
Integrative Omics Analysis Reveals Mechanisms of Anthocyanin Biosynthesis in Djulis Spikes
by Chunmei Zheng, Wenxuan Ge, Xueying Li, Xiuzhang Wang, Yanxia Sun and Xiaoyong Wu
Plants 2025, 14(2), 197; https://doi.org/10.3390/plants14020197 - 12 Jan 2025
Viewed by 709
Abstract
Djulis (Chenopodium formosanum Koidz.), a member of the Amaranthaceae family plant, is noted for its vibrant appearance and significant ornamental value. However, the mechanisms underlying color variation in its spikes remain unexplored. This research initially detected the anthocyanin content at different developmental [...] Read more.
Djulis (Chenopodium formosanum Koidz.), a member of the Amaranthaceae family plant, is noted for its vibrant appearance and significant ornamental value. However, the mechanisms underlying color variation in its spikes remain unexplored. This research initially detected the anthocyanin content at different developmental stages of the spike and subsequently utilized an integrative approach, combining targeted metabolomics, transcriptomics, and untargeted metabolomics analyses, to elucidate the mechanisms of anthocyanin biosynthesis in the spikes of djulis. The results of the combined multi-omics analysis showed that the metabolites associated with anthocyanin synthesis were mainly enriched in the flavonoid biosynthesis pathway (ko00941) and the anthocyanin biosynthesis pathway (ko00942). With the maturation of djulis spikes, a total of 28 differentially expressed genes and 17 differentially expressed metabolites were screened during the transition of spike color from green (G) to red (R) or orange (O). Twenty differentially expressed genes were selected for qRT-PCR validation, and the results are consistent with transcriptome sequencing. The upregulation of seven genes, including chalcone synthase (CfCHS3_1, CfCHS3_2, CfCHS3_3), flavanone 3-hydroxylase (CfF3H_3), flavonoid 3′5′-hydroxylase (CfCYP75A6_1), dihydroflavonol reductase (CfDFRA), and glucosyltransferase (Cf3GGT), promotes the formation and accumulation of delphinidin 3-sambubioside and peonidin 3-galactoside. The research results also showed that anthocyanins and betalains can coexist in the spike of djulis, and the reason for the change in spike color during development may be the result of the combined action of the two pigments. A possible regulatory pathway for anthocyanin biosynthesis during the spike maturation was constructed based on the analysis results. The results provide a reference and theoretical basis for further studying the molecular mechanism of anthocyanin regulation of color changes in Amaranthaceae plants. Full article
(This article belongs to the Special Issue Horticultural Plant Physiology and Molecular Biology)
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<p>Anthocyanin biosynthesis pathway. PAL: phenylalanine ammonia-lyase; C4H: cinnamate-4-hydroxylase; 4CL: 4-coumarate-CoA ligase; CHS: chalcone synthase; CHI: chalcone isomerase; F3H: flavanone 3-hydroxylase; F3′H: flavonoid 3′-hydroxylase; F3′5′H: flavonoid 3′,5′-hydroxylase; DFR: dihydroflavonol 4-reductase; ANS: anthocyanidin synthase; LDOX: leucoanthocyanidin dioxygenase; UFGT: flavonoid-glucosyl transferase; MT: methyltransferase; GT: glucosyltransferase; AT: acyltransferase.</p>
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<p>Detection of anthocyanin content. (<b>A</b>–<b>G</b>) Seven stages of spike development in djulis. The labels (<b>A</b>–<b>G</b>) represent seven developmental phases, specifically from the full bloom stage to 15, 20, 25, 30, 35, and 40 days after pollination (DAP). (<b>H</b>) Line graph of changes in anthocyanin content in djulis spikes at different developmental stages.</p>
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<p>Targeted metabolomics analysis. (<b>A</b>) Pie chart of anthocyanin full identification classification.by targeted metabolome. (<b>B</b>) Different anthocyanin quantities in different comparison combinations (<b>C</b>) Heatmap of O vs. G differential anthocyanin hierarchical clustering. (<b>D</b>) Heatmap of R vs. G differential anthocyanin hierarchical clustering.</p>
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<p>Non-targeted metabolomics analysis. (<b>A</b>) PCA score plot in positive ion mode. (<b>B</b>) PCA score plot in negative ion mode. (<b>C</b>) The number of differential metabolites in each comparison combination under positive ion mode. (<b>D</b>) The number of differential metabolites in each comparison combination under negative ion mode.</p>
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<p>Transcriptomics analysis. The red box in the figure represents the flavonoid biosynthesis. (<b>A</b>) Differential gene Wayne plots. (<b>B</b>) O vs. G KEGG enrichment scatter plot. (<b>C</b>) R vs. G KEGG enrichment scatter plot. (<b>D</b>) R vs. O KEGG enrichment scatter plot.</p>
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<p>Heatmap of the correlation between differential genes and differential anthocyanin metabolites. (<b>A</b>) O versus G. (<b>B</b>) R versus G.</p>
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<p>Line graph of real-time PCR analysis of anthocyanin biosynthesis-related genes in the spike of djulis. 1–4 represent the 20th, 30th, 35th, and 40th days after the end of the full flowering period, respectively. A B, and C represent three biological replicates. (<b>A</b>) <span class="html-italic">CfCHS_1</span>(<span class="html-italic">Cf062520</span>, chalcone synthase). (<b>B</b>) <span class="html-italic">CfCHS_2</span>(<span class="html-italic">Cf001167</span>, chalcone synthase). (<b>C</b>) <span class="html-italic">CfCHS_3</span>(<span class="html-italic">Cf001178</span>, chalcone synthase). (<b>D</b>) <span class="html-italic">CfCHS_4</span>(<span class="html-italic">Cf062524</span>, chalcone synthase). (<b>E</b>) <span class="html-italic">CfCHS3_3</span>(<span class="html-italic">Cf076825</span>, chalcone synthase 3). (<b>F</b>) <span class="html-italic">CfCHI2_1</span>(<span class="html-italic">Cf054781</span>, chalcone isomerase). (<b>G</b>) <span class="html-italic">CfCHI2_2</span>(<span class="html-italic">Cf022835</span>, chalcone isomerase). (<b>H</b>) <span class="html-italic">CfF3H_2</span>(<span class="html-italic">Cf016292</span>, flavanone 3-hydroxylase). (<b>I</b>) <span class="html-italic">CfF3H_3</span>(<span class="html-italic">Cf040556</span>, flavanone 3-hydroxylase). (<b>J</b>) <span class="html-italic">CfCYP75B2_1</span>(<span class="html-italic">Cf034387</span>, flavonoid 3′-monooxygenase). (<b>K</b>) <span class="html-italic">CfCYP75B2_2</span>(<span class="html-italic">Cf058394</span>, flavonoid 3′-monooxygenase). (<b>L</b>) <span class="html-italic">CfCYP75A6_1</span>(<span class="html-italic">Cf006386</span>, flavonoid 3′,5′-hydroxylase). (<b>M</b>) <span class="html-italic">CfCYP75A6_2</span>(<span class="html-italic">Cf032051</span>, flavonoid 3′,5′-hydroxylase). (<b>N</b>) <span class="html-italic">CfDFRA</span>(<span class="html-italic">Cf074499</span>, dihydroflavonol 4-reductase). (<b>O</b>) <span class="html-italic">CfANS</span>(<span class="html-italic">Cf010560</span>, anthocyanidin synthase). (<b>P</b>) <span class="html-italic">CfLDOX</span>(<span class="html-italic">Cf061103</span>, leucoanthocyanidin dioxygenase). (<b>Q</b>) <span class="html-italic">CfUFGT_1</span>(<span class="html-italic">Cf015504</span>, anthocyanidin 3-O-glucosyltransferase). (<b>R</b>) <span class="html-italic">CfUFGT_3</span>(<span class="html-italic">Cf015507</span>, anthocyanidin 3-O-glucosyltransferase). (<b>S</b>) <span class="html-italic">Cf3GGT</span> (<span class="html-italic">Cf005087</span>, anthocyanidin 3-O-glucoside 2″-O-glucosyltransferase).</p>
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<p>Anthocyanin biosynthesis pathway in the spike of djulis. PAL: phenylalanine ammonialyase; C4H: cinnamate-4-hydroxylase; 4CL: 4-Coumarate-CoA ligase; CHS: chalcone synthase; CHI: chalcone isomerase; F3H: flavanone 3-hydroxylase; F3′H: flavonoid 3′-hydroxylase; F3’5’H: flavonoid 3’,5’-hydroxylase; DFR: dihydroflavonol 4-reductase; ANS: anthocyanidin synthase; LDOX: leucoanthocyanidin dioxygenase; UFGT: UDP-glucose flavonoid 3-O-glucosyl transferase; GT: glucosyltransferase.</p>
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<p>Djulis in different color periods. (<b>A</b>) The spike of djulis during the green period before maturity. (<b>B</b>) The spike of djulis during the red period after maturity. (<b>C</b>) The spike of djulis during the orange period after maturity.</p>
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21 pages, 2156 KiB  
Article
Valorization of Betalain Pigments Extracted from Phytolacca americana L. Berries as Natural Colorant in Cheese Formulation
by Ionuț Dumitru Veleșcu, Ioana Cristina Crivei, Andreea Bianca Balint, Vlad Nicolae Arsenoaia, Alexandru Dragoș Robu, Florina Stoica and Roxana Nicoleta Rațu
Agriculture 2025, 15(1), 86; https://doi.org/10.3390/agriculture15010086 - 2 Jan 2025
Viewed by 683
Abstract
In response to consumer demand for more sustainable and health-conscious products, the food sector is increasingly shifting towards the use of natural additives. Pokeweed (Phytolacca americana L.) is a medicinal plant that contains valuable biologically active compounds, including betacyanins, which serve as [...] Read more.
In response to consumer demand for more sustainable and health-conscious products, the food sector is increasingly shifting towards the use of natural additives. Pokeweed (Phytolacca americana L.) is a medicinal plant that contains valuable biologically active compounds, including betacyanins, which serve as its red pigments, along with phenolic acids, flavonoids, polyphenolic compounds, and others. Phytolacca americana (P. americana) is a plant renowned for its bioactive compounds, which exhibit anti-inflammatory, anti-mutagenic, antioxidant, anticancer, and antibacterial properties. This study investigates the potential of betalain pigments extracted from the berries of P. americana as a natural colorant for cheese formulation. The impact of these pigments on the color attributes, sensory qualities, and physicochemical and phytochemical composition of the cheeses was systematically evaluated. The Phytolacca americana (PA) powder demonstrated significant levels of total polyphenols (111.95 ± 1.60 mg GAE/g dw) and antioxidant activity (21.67 ± 0.19 µmol TE/g dw). The incorporation of PA powder increased the physicochemical and phytochemical contents and antioxidant activity in the final product (4.40 ± 0.22 µmol TE/g dw for CPAP1 and 6.11 ± 0.22 µmol TE/g dw for CPAP2). The sensory study revealed that the PA-supplemented cheeses were acceptable. The enhanced cheeses present a distinctive color profile, attracting health-conscious consumers looking for innovative dairy products. The study concludes that PA powder can effectively enhance cheese, producing a phytochemical-enriched product that appeals to health-conscious consumers. Full article
(This article belongs to the Special Issue Quality Assessment and Processing of Farm Animal Products)
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<p>(<b>A</b>) <span class="html-italic">P. americana</span> plant, (<b>B</b>) <span class="html-italic">P. americana</span> fruits, and (<b>C</b>) <span class="html-italic">P. americana</span> powder.</p>
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<p>Technological flow of cheese manufacturing.</p>
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<p>Spider diagrams corresponding to the descriptive sensory analysis of the control and enhanced semi-hard cheeses (control (CC), cheese with 1% PA (CPAP1), and cheese with 2% PA (CPAP2)).</p>
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<p>Principal components analysis (PCA) of the sensory attributes of the control and enhanced semi-hard cheeses (control (CC), cheese with 1% PA (CPAP1), and cheese with 2% PA (CPAP2)).</p>
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<p>Images of the control and enhanced semi-hard cheeses. (<b>a</b>) Control (CC), (<b>b</b>) cheese with 1%PA (CPAP1), and (<b>c</b>) cheese with 2%PA (CPAP2).</p>
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17 pages, 2268 KiB  
Article
Anti-Inflammatory Activity of Gomphrenin-Rich Fraction from Basella alba L. f. rubra Fruits
by Agnieszka Rusak, Monika Mrozowska, Weronika Kozłowska, Benita Wiatrak, Piotr Dzięgiel, Sławomir Wybraniec, Ivana Carev, Agnieszka Jama-Kmiecik, Adam Matkowski and Sylwia Zielińska
Nutrients 2024, 16(24), 4393; https://doi.org/10.3390/nu16244393 - 20 Dec 2024
Viewed by 808
Abstract
Background/Objectives: Basella alba L. (Malabar spinach, Basellaceae), widely consumed as a leafy vegetable, produces dark-colored fruits rich in betacyanins, including rare 6-glycosylated derivatives called gomphrenins. Comprehensive studies on the anti-inflammatory potential of its gomphrenin fraction (A) and crude extract (B) employed various [...] Read more.
Background/Objectives: Basella alba L. (Malabar spinach, Basellaceae), widely consumed as a leafy vegetable, produces dark-colored fruits rich in betacyanins, including rare 6-glycosylated derivatives called gomphrenins. Comprehensive studies on the anti-inflammatory potential of its gomphrenin fraction (A) and crude extract (B) employed various analytical and biological methods. Methods: Cytotoxicity and anti-inflammatory effects were tested on human and animal cell models using SRB, DCF-DA, Griess, MDA, and ELISA assays. Results: Both the crude extract and enriched gomphrenin fraction exhibited significant anti-inflammatory and antioxidant effects in vitro. They inhibited pro-inflammatory cytokines IL-1β and IL-6, reduced oxidative stress markers (ROS, NO), and decreased lipid peroxidation. The enriched gomphrenin fraction (A) showed stronger antioxidant and anti-inflammatory effects, particularly in reducing ROS and NO levels, though not always concentration-dependent. Both A and B inhibited NF-κB and COX activity. Conclusions: These findings highlight the therapeutic potential of B. alba f. rubra fruit extract and betalain fraction for inflammation-related conditions, encouraging further exploration of their mechanisms and application. Full article
(This article belongs to the Section Phytochemicals and Human Health)
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<p>Chemical structures of gomphrenin (glycosylated at carbon 6) and betanin (more commonly found glycosylation at carbon 5).</p>
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<p>L929 cell line SRB test for gomphrenin enriched fraction (<b>A</b>) and crude extract (<b>B</b>). KK—cell culture in a culture medium; KR—culture fluid without the presence of cells. Statistical significance of differences was estimated using the Kruskal–Wallis test at <span class="html-italic">p</span> &lt; 0.05 and is indicated on the figure—the same letters indicate lack of statistical significance of differences.</p>
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<p>NHDF cell line SRB test for gomphrenin enriched fraction (<b>A</b>) and crude extract (<b>B</b>). KK—cell culture in a culture medium; KR—culture fluid without the presence of cells. Statistical significance of differences was estimated using the Kruskal–Wallis test at <span class="html-italic">p</span> &lt; 0.05 (*) (the same lowercase letters above the bars indicate lack of significant differences).</p>
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<p>(<b>a</b>–<b>d</b>) The effect of betalain enriched fraction (A), crude extract (B), meloxicam (Mel), indomethacin (Ind), ascorbic acid (Asc), aspirin (Asp), and lipopolysaccharide (LPS) on ROS secretion in THP-1 cell model using DCF-DA assay (<b>a</b>); MDA assay (<b>b</b>); Griess assay (<b>c</b>); COX activity assay (<b>d</b>) with different bar shading indicating COX-1 and COX -2 isoforms and non-specific activity. Statistical significance of differences was estimated using the Kruskal–Wallis test at <span class="html-italic">p</span> &lt; 0.05 (*).</p>
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<p>(<b>a</b>–<b>c</b>) The effect of enriched gomphrenin fraction (A) and crude extract on cytokines IL-1β (<b>a</b>) and IL-6 (<b>b</b>) and transcription factor NF-κB (<b>c</b>) secretion in LPS-stimulated THP-1 cell model. Meloxicam (Mel), indomethacin (Ind), ascorbic acid (Asc), medium (M). Statistical significance of differences was estimated using the Kruskal–Wallis test at <span class="html-italic">p</span> &lt; 0.05 (*).</p>
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31 pages, 12442 KiB  
Article
Poly (Lactic Acid) Fibrous Film with Betalains from Pitaya (Stenocereus thurberi) by Electrospinning for Potential Use as Smart Food Packaging
by Dalila Fernanda Canizales-Rodríguez, Francisco Rodríguez-Félix, José Agustín Tapia-Hernández, Carmen Lizette Del-Toro-Sánchez, Saúl Ruíz-Cruz, Santiago P. Aubourg, Victor Manuel Ocaño-Higuera, María Irene Silvas-García, Cielo Estefanía Figueroa-Enríquez and Milagros Guadalupe Álvarez-Moreno
Coatings 2024, 14(12), 1581; https://doi.org/10.3390/coatings14121581 - 18 Dec 2024
Viewed by 1038
Abstract
The incorporation of biopolymers and natural colorants in smart packaging has garnered significant attention in the food packaging industry. This study investigates the design and characterization of novel fibrous films incorporating betalain extract (BE) from Stenocereus thurberi in poly (lactic acid) (PLA). An [...] Read more.
The incorporation of biopolymers and natural colorants in smart packaging has garnered significant attention in the food packaging industry. This study investigates the design and characterization of novel fibrous films incorporating betalain extract (BE) from Stenocereus thurberi in poly (lactic acid) (PLA). An electrospinning technique was developed with varying PLA concentrations (2%–12% w/v) and BE concentrations (8%–12% w/v) to create a colorimetric freshness indicator. BE was characterized by quantifying its phytochemical content and assessing its antioxidant capacity. Morphological and structural analyses included scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FTIR), polydispersity index (PI), mechanical properties, and functional characteristics such as ammonia sensitivity and total antioxidant activity. The results indicated that the incorporation of BE significantly influenced the average diameter of the nanofibers, ranging from 313 ± 74 nm to 657 ± 99 nm. SEM micrographs showed that PLA12-BE12 films exhibited smooth surfaces without bead formation. The FTIR analysis confirmed effective BE incorporation, revealing intermolecular interactions between the betalain molecules and the PLA matrix, which contributed to enhanced structural and functional stability. The mechanical properties analysis revealed that moderate BE additions (8%–10% w/v) enhanced the Young’s modulus and tensile strength, while higher BE concentrations (12% w/v) disrupted the polymer network, reducing these properties. Additionally, the strain at break decreased significantly with BE incorporation, reflecting limited molecular chain mobility. Increasing BE concentration notably improved antioxidant activity, with the BE concentration of 12% (w/v), the ABTS•+, DPPH•, and FRAP radical scavenging activities at the highest values of 84.28 ± 1.59%, 29.95 ± 0.34%, and 710.57 ± 28.90 µM ET/g, respectively. Ammonia sensitivity tests demonstrated a significant halochromic transition from reddish-pink to yellow, indicating high sensitivity to low ammonia concentrations. The possible mechanism is alkaline pH induces aldimine bond hydrolysis and generates betalamic acid (yellow) and cyclo-DOPA-5-O-ß-glucoside (colorless) The fibrous films also exhibited reversible color changes and maintained good color stability over 30 days, emphasizing their potential for use in smart packaging applications for real-time freshness monitoring and food quality assessment. Full article
(This article belongs to the Special Issue Novel Advances in Food Contact Materials)
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<p>Schematic diagram of the production of PLA12-BE12 nanofibrous film by electrospinning technique.</p>
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<p>UV-vis spectra (<b>a</b>,<b>b</b>) and photograph of color variations (<b>c</b>) in pH 1–14 range of betalain solutions.</p>
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<p>SEM micrographs of PLA films at 2 (<b>a</b>), 4 (<b>b</b>), 6 (<b>c</b>), 8 (<b>d</b>), 10 (<b>e</b>), and 12 (<b>f</b>) % (<span class="html-italic">w</span>/<span class="html-italic">v</span>) fiber. Magnifications images are 1000× and 5000×. Yellow circles show fibers defects.</p>
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<p>SEM micrographs of PLA 8, 10, and 12% (<span class="html-italic">w</span>/<span class="html-italic">v</span>) fibrous film incorporated with BE at 8% (<span class="html-italic">w</span>/<span class="html-italic">v</span>). The magnification images are 1000× and 5000×. Yellow circles show fibers defects.</p>
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<p>SEM micrographs of PLA 8, 10, and 12% (<span class="html-italic">w</span>/<span class="html-italic">v</span>) film incorporated with BE at 10% (<span class="html-italic">w</span>/<span class="html-italic">v</span>). The magnification images are 1000× and 5000×. Yellow circles show fibers defects.</p>
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<p>SEM micrographs of PLA 8, 10, and 12% (<span class="html-italic">w</span>/<span class="html-italic">v</span>) fibrous films incorporated with BE at 12% (<span class="html-italic">w</span>/<span class="html-italic">v</span>). The magnification images are 1000× and 5000×. Yellow circles show fibers defects.</p>
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<p>SEM micrographs of PLA 12% (<b>a</b>), PLA12%-BE8% (<b>b</b>), PLA12%-BE10% (<b>c</b>), and PA12%-BE12% (<b>d</b>) (<span class="html-italic">w</span>/<span class="html-italic">v</span>) fibrous films and their corresponding histograms. The magnification images are 5000×.</p>
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<p>Physical appearance and color value (<span class="html-italic">L</span>, <span class="html-italic">a</span>*, <span class="html-italic">b</span>* and Δ<span class="html-italic">E</span>) of PLA 8, 10, and 12% (<span class="html-italic">w</span>/<span class="html-italic">v</span>) fibrous films with the addition of BE at 8, 10, and 12% (<span class="html-italic">w</span>/<span class="html-italic">v</span>).</p>
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<p>FTIR spectra of the fibrous film: PLA 12% (<b>a</b>), BE (<b>b</b>), PLA12-BE12 (<b>c</b>).</p>
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<p>Color response of PLA12-BET12 indicator fibrous film at different ammonia concentrations: photographs after reaction with ammonia (<b>a</b>) and color values of the film after reaction with ammonia (Δ<span class="html-italic">E</span>, Δ<span class="html-italic">L</span>, Δ<span class="html-italic">a</span>, Δ<span class="html-italic">b</span>) (<b>b</b>) at 10 min of reaction. Data with different letters on the same column color refer to significant differences (<span class="html-italic">p</span> &lt; 0.05). Data (mean ± SD) are from three independent experiments.</p>
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<p>Color response of PLA12-BE12 fibrous film, at different ammonia concentrations: photographs after reaction with ammonia (<b>a</b>), color values of the film after reaction with ammonia (Δ<span class="html-italic">E</span>, Δ<span class="html-italic">L</span>, Δ<span class="html-italic">a</span>, Δ<span class="html-italic">b</span>) (<b>b</b>), at 20 min of reaction. Data with different letters on the same column color reference to significant differences (<span class="html-italic">p</span> &lt; 0.05). Data (mean ± SD) are from three independent experiments.</p>
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<p>ABTS•+ radical inhibition (<b>a</b>), DPPH• radical inhibition (<b>b</b>), and FRAP µM TE/g (<b>c</b>) of different fibrous films at various immersion times. Data (mean ± SD) are from three independent experiments.</p>
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<p>Color responses of PLA12-BE12 fibrous film after reaction during four cycles in extremely acidic and alkaline atmospheres.</p>
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<p>Total color change (Δ<span class="html-italic">E</span>) for PLA12-BE12 fibrous film during storage at 4 °C and 25 °C at 30 days. Data with different letters on the same column color refer to significant differences (<span class="html-italic">p</span> &lt; 0.05). Data (mean ± SD) are from three independent experiments.</p>
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<p>Photograph of PLA12-BE12 intelligent fibrous film indicator during storage at 4 °C and 25 °C at 30 days.</p>
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<p>Hue angles of the intelligent fibrous film indicator during storage at 4 °C and 25 °C at 30 days. Data with different letters on the same column color refer to significant differences (<span class="html-italic">p</span> &lt; 0.05). Data (mean ± SD) are from three independent experiments.</p>
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<p>Chroma values of the indicator fibrous film during storage at 4 °C and 25 °C at 30 days. Data with different letters on the same column color refer to significant differences (<span class="html-italic">p</span> &lt; 0.05). Data (mean ± SD) are from three independent experiments.</p>
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