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19 pages, 13434 KiB  
Article
Identifying Candidate Genes Related to Soybean (Glycine max) Seed Coat Color via RNA-Seq and Coexpression Network Analysis
by Cheng Wang, Pingchun Fu, Tingting Sun, Yan Wang, Xueting Li, Shulin Lan, Hui Liu, Yongji Gou, Qiaoxia Shang and Weiyu Li
Genes 2025, 16(1), 44; https://doi.org/10.3390/genes16010044 - 1 Jan 2025
Viewed by 453
Abstract
Background: The quality of soybeans is reflected in the seed coat color, which indicates soybean quality and commercial value. Researchers have identified genes related to seed coat color in various plants. However, research on the regulation of genes related to seed coat color [...] Read more.
Background: The quality of soybeans is reflected in the seed coat color, which indicates soybean quality and commercial value. Researchers have identified genes related to seed coat color in various plants. However, research on the regulation of genes related to seed coat color in soybeans is rare. Methods: In this study, four lines of seed coats with different colors (medium yellow 14, black, green, and brown) were selected from the F2:5 population, with Beinong 108 as the female parent and green bean as the male parent, and the dynamic changes in the anthocyanins in the seed coat were stained with 4-dimethylaminocinnamaldehyde (DMACA) during the grain maturation process (20 days from grain drum to seed harvest). Through RNA-seq of soybean lines with four different colored seed coats at 30 and 50 days after seeding, we can further understand the key pathways and gene regulation modules between soybean seed coats of different colors. Results: DMACA revealed that black seed coat soybeans produce anthocyanins first and have the deepest staining. Clustering and principal component analysis (PCA) of the RNA-seq data divided the eight samples into two groups, resulting in 16,456 DEGs, including 5359 TFs. GO and KEGG enrichment analyses revealed that the flavonoid biosynthesis, starch and sucrose metabolism, carotenoid biosynthesis, and circadian rhythm pathways were significantly enriched. We also conducted statistical and expression pattern analyses on the differentially expressed transcription factors. Based on weighted gene coexpression network analysis (WGCNA), we identified seven specific modules that were significantly related to the four soybean lines with different seed coat colors. The connectivity and functional annotation of genes within the modules were calculated, and 21 candidate genes related to soybean seed coat color were identified, including six transcription factor (TF) genes and three flavonoid pathway genes. Conclusions: These findings provide a theoretical basis for an in-depth understanding of the molecular mechanisms underlying differences in soybean seed coat color and provide new genetic resources. Full article
(This article belongs to the Special Issue Abiotic Stress in Plants: Genetics and Genomics)
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<p>Four different soybean seed coat colors after 30 days and 50 days of drumming.</p>
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<p>DMACA staining images of the seed coat bulging into four colors from 20 d to 54 d. (<b>a</b>) The yellow seed coat is unstained, (<b>b</b>) the yellow seed coat is stained, (<b>c</b>) the green seed coat is unstained, (<b>d</b>) the green seed coat is stained, (<b>e</b>) the brown seed coat is unstained, (<b>f</b>) the brown seed coat is stained, (<b>g</b>) the black seed coat is unstained, and (<b>h</b>) the black seed coat is stained.</p>
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<p>(<b>a</b>) Cluster dendrogram showing the two different developmental stages of the dynamics of changes in the color of the soybean seed coat. (<b>b</b>) PCA of eight RNA-seq samples.</p>
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<p>(<b>a</b>) Number of differentially expressed genes in different lines. (<b>b</b>) Venn diagram of differentially expressed genes within different lines. (<b>c</b>) GO enrichment analysis of differentially expressed genes within the lines. (<b>d</b>) KEGG enrichment analysis of differentially expressed genes within the lines.</p>
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<p>Line chart of DEG expression patterns from different developmental periods (30 d vs. 50 d) of the same lines.</p>
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<p>(<b>a</b>) UpSet plots of differentially expressed genes from the same developmental period of different lines. (<b>b</b>) GO enrichment analysis of DEGs in the same developmental period of different lines. (<b>c</b>) KEGG enrichment analysis of DEGs in the same developmental period of different lines.</p>
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<p>Line chart of DEG expression patterns for the same developmental period of different lines.</p>
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<p>(<b>a</b>) Treemap of the percentage of TFs within the line. (<b>b</b>) Clustering heatmap of TFs within the line. The right side shows the number of TFs in the top five clusters.</p>
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<p>(<b>a</b>) Hierarchical clustering tree for gene coexpression network analysis. (<b>b</b>) Heatmap of correlations and significance between modules and different samples. (<b>c</b>) Gene coexpression network module; different colors were used to represent different modules.</p>
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<p>(<b>a</b>) qRT–PCR was used to generate a heatmap of the expression patterns of 21 candidate genes in different seed coat color lines. qRT–PCR was used to generate a heatmap of the expression patterns of 21 candidate genes in different seed coat color lines. (<b>b</b>) Scatter plot of RNA-seq and qRT–PCR correlations.</p>
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15 pages, 3459 KiB  
Article
Isolation of a Novel Low-Temperature-Active and Organic-Solvent-Stable Mannanase from the Intestinal Metagenome of Hermetia illucens
by Dong-Gwan Kim, Chang-Muk Lee, Young-Seok Lee, Sang-Hong Yoon and Su-Yeon Kim
Int. J. Mol. Sci. 2025, 26(1), 216; https://doi.org/10.3390/ijms26010216 - 30 Dec 2024
Viewed by 290
Abstract
The black soldier fly, Hermetia illucens, is a voracious scavenger of various organic materials; therefore, it could be exploited as a biological system for processing daily food waste. In order to survey novel hydrolytic enzymes, we constructed a fosmid metagenome library using [...] Read more.
The black soldier fly, Hermetia illucens, is a voracious scavenger of various organic materials; therefore, it could be exploited as a biological system for processing daily food waste. In order to survey novel hydrolytic enzymes, we constructed a fosmid metagenome library using unculturable intestinal microorganisms from H. illucens. Through functional screening of the library on carboxymethyl cellulose plates, we identified a fosmid clone, the product of which displayed hydrolytic activity. Sequence analysis of the fosmid revealed a novel mannan-degrading gene, ManEM6, composed of 1185 base pairs encoding 394 amino acids, with a deduced 20-amino-acid N-terminal signal peptide sequence. The conceptual translation of ManEM6 exhibited the highest identity (78%) to endo-1,4-β-mannosidase from Dysgonomonas mossii. Phylogenetic and domain analyses indicated that ManEM6 encodes a novel mannanase with a glycoside hydrolase family 26 domain. The recombinant protein rManEM6 showed its highest activity at 40 °C and pH 7.0, and it remained stable in the range of pH 5–10.0. rManEM6 hydrolyzed substrates with β-1,4-glycosidic mannoses, showing maximum enzymatic activity toward locust bean gum galactomannan, while it did not hydrolyze p-nitrophenyl-β-pyranosides, demonstrating endo-form mannosidase activity. rManEM6 was highly stable under stringent conditions, including those of polar organic solvents, as well as reducing and denaturing reagents. Therefore, ManEM6 may be an attractive candidate for the degradation of mannan under high-organic-solvent and protein-denaturing processes in the food and feed industries. Full article
(This article belongs to the Section Molecular Informatics)
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<p>Multiple alignment and phylogenetic tree of ManEM6. (<b>a</b>) Multiple sequence alignment with glycoside hydrolase family 26 (GH26): β-endo-mannanase showed three putative catalytic residues (Glu189, Asp254, and Glu296), which are shown in dark-shaded letters. Asterisks (*) indicate consensus amino acids. Each protein sequence used for alignment was collected from GenBank, including <span class="html-italic">Bacillus</span> sp. JAMB750 [<a href="#B29-ijms-26-00216" class="html-bibr">29</a>], <span class="html-italic">Bacillus subtilis</span> [<a href="#B31-ijms-26-00216" class="html-bibr">31</a>], <span class="html-italic">Cellvibrio japonicas</span> [<a href="#B32-ijms-26-00216" class="html-bibr">32</a>], <span class="html-italic">Bacteroides ovatus</span> [<a href="#B33-ijms-26-00216" class="html-bibr">33</a>], <span class="html-italic">Bacteroides</span> sp. 2_1_33B [<a href="#B34-ijms-26-00216" class="html-bibr">34</a>]. (<b>b</b>) The phylogenetic tree of ManEM6 and other closely related enzymes was reconstructed using the neighbor-joining method (MEGA5.1 software). The protein sequences of related enzymes were retrieved from NCBI GenBank. Bootstrap values (&gt;50%) at the nodes were based on 1000 replicates of the dataset. Solid circles indicate that the corresponding branches were also recovered in both the maximum parsimony and maximum likelihood trees. Bar, 0.2 substitutions per amino acid position.</p>
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<p>Multiple alignment and phylogenetic tree of ManEM6. (<b>a</b>) Multiple sequence alignment with glycoside hydrolase family 26 (GH26): β-endo-mannanase showed three putative catalytic residues (Glu189, Asp254, and Glu296), which are shown in dark-shaded letters. Asterisks (*) indicate consensus amino acids. Each protein sequence used for alignment was collected from GenBank, including <span class="html-italic">Bacillus</span> sp. JAMB750 [<a href="#B29-ijms-26-00216" class="html-bibr">29</a>], <span class="html-italic">Bacillus subtilis</span> [<a href="#B31-ijms-26-00216" class="html-bibr">31</a>], <span class="html-italic">Cellvibrio japonicas</span> [<a href="#B32-ijms-26-00216" class="html-bibr">32</a>], <span class="html-italic">Bacteroides ovatus</span> [<a href="#B33-ijms-26-00216" class="html-bibr">33</a>], <span class="html-italic">Bacteroides</span> sp. 2_1_33B [<a href="#B34-ijms-26-00216" class="html-bibr">34</a>]. (<b>b</b>) The phylogenetic tree of ManEM6 and other closely related enzymes was reconstructed using the neighbor-joining method (MEGA5.1 software). The protein sequences of related enzymes were retrieved from NCBI GenBank. Bootstrap values (&gt;50%) at the nodes were based on 1000 replicates of the dataset. Solid circles indicate that the corresponding branches were also recovered in both the maximum parsimony and maximum likelihood trees. Bar, 0.2 substitutions per amino acid position.</p>
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<p>Overexpression and purification of rManEM6. ManEM6 without putative <span class="html-italic">N</span>-terminal signal peptide sequence was cloned into plasmid pET21a (+) expression vector and expressed in <span class="html-italic">E. coli</span> BL21(DE3). SDS–PAGE analysis shows a homogeneous 43 kDa protein. Lanes: M, molecular weight marker; Lane 1, total cellular protein from uninduced cells; Lane 2, induced total cellular protein; Lane 3, induced soluble fraction; Lane 4, purified rManEM6 protein; Lane 5, zymogram of rManEM6 on polyacrylamide gel with 0.2% LBG (locust bean gum) and stained with Congo-red.</p>
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<p>Biochemical characterization of rManEM6. (<b>a</b>) The enzyme activity of rManEM6 (0.2 μg) was measured at various temperatures in 50 mM sodium phosphate (pH 7.0) for 10 min, exhibiting &gt;50% hydrolytic activity in the range of 25–50 °C with an optimum at 40 °C. (<b>b</b>) The thermostability of rManEM6 was examined at 40 °C for 10 min after 4 h pre-incubation at the given temperature. (<b>c</b>) pH inhibition was monitored in the following 50 mM buffers: sodium acetate buffer (NaOAc, pH 5.0–6.0), sodium phosphate buffer (Na-Pi, pH 6.0–7.5), 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid buffer (HEPES, pH 7.5–8.5), and 2-(cyclohexylamino) ethanesulfonic acid buffer (CHES, pH 8.5–10.0). (<b>d</b>) To verify pH stability, the enzyme was pre-incubated in 50 mM buffer with different pH values at 4 °C for 15 h, and residual activity was examined. Each β-mannanase assay was carried out by measuring the amount of mannose released from locust bean gum (0.5% [<span class="html-italic">w</span>/<span class="html-italic">v</span>]) under standard assay conditions, as described in the Materials and Methods section. Error bars represent SEM from triplicate results (<span class="html-italic">p</span> ≤ 0.01).</p>
Full article ">Figure 3 Cont.
<p>Biochemical characterization of rManEM6. (<b>a</b>) The enzyme activity of rManEM6 (0.2 μg) was measured at various temperatures in 50 mM sodium phosphate (pH 7.0) for 10 min, exhibiting &gt;50% hydrolytic activity in the range of 25–50 °C with an optimum at 40 °C. (<b>b</b>) The thermostability of rManEM6 was examined at 40 °C for 10 min after 4 h pre-incubation at the given temperature. (<b>c</b>) pH inhibition was monitored in the following 50 mM buffers: sodium acetate buffer (NaOAc, pH 5.0–6.0), sodium phosphate buffer (Na-Pi, pH 6.0–7.5), 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid buffer (HEPES, pH 7.5–8.5), and 2-(cyclohexylamino) ethanesulfonic acid buffer (CHES, pH 8.5–10.0). (<b>d</b>) To verify pH stability, the enzyme was pre-incubated in 50 mM buffer with different pH values at 4 °C for 15 h, and residual activity was examined. Each β-mannanase assay was carried out by measuring the amount of mannose released from locust bean gum (0.5% [<span class="html-italic">w</span>/<span class="html-italic">v</span>]) under standard assay conditions, as described in the Materials and Methods section. Error bars represent SEM from triplicate results (<span class="html-italic">p</span> ≤ 0.01).</p>
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<p>Enzymatic stability of rManEM6 against enzyme inhibitors. (<b>a</b>) The same amount of rManEM6 protein (0.2 μg) was pre-incubated at 30 °C for 30 min with different organic solvents (10% [<span class="html-italic">v</span>/<span class="html-italic">v</span>]) in 50 mM sodium phosphate buffer (pH 7.0, 200 μL), and residual β-mannanase activity was measured at 40 °C for 10 min with locust bean gum (0.5%, [<span class="html-italic">w</span>/<span class="html-italic">v</span>]) as a substrate. The bar represents the log-polarity of each organic solvent. DMSO, dimethyl sulfoxide; DMF, dimethylformamide. (<b>b</b>) To determine stability under chemical reagents, rManEM6 (0.2 μg protein) was pre-incubated with various denaturants or reducing agents at 30 °C for 30 min, and residual activity was measured. Gu-HCl, guanidine hydrochloride; gu-thiocyanate, guanidine thiocyanate. (<b>c</b>) An enzyme assay was performed after pre-incubation of rManEM6 with 1 mM of each of the indicated metal ions at 30 °C for 30 min. The relative amount of mannose produced was compared with that in the standard reaction. (<b>d</b>) To examine the effects of detergents on rManEM6 stability, the residual activity from pre-incubated enzyme (in 10% ([<span class="html-italic">v</span>/<span class="html-italic">v</span>] or [<span class="html-italic">v</span>/<span class="html-italic">w</span>]) detergent at 30 °C for 30 min) was measured. SDS, sodium dodecyl sulfate; CTAB, cetyltrimethylammonium bromide. The activity measured in the absence of any of the chemical compounds is shown as 100% (<span class="html-italic">p</span> ≤ 0.01). ND, not detectable.</p>
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<p>Thin layer chromatography analysis of hydrolysis products of mannose oligo- and polysaccharides by rManEM6. Enzymatic hydrolysis of each substrate (0.5%, <span class="html-italic">w</span>/<span class="html-italic">v</span>) was performed either with (+lanes) or without (–lanes) ManEM6 in 50 mM sodium phosphate buffer (pH 7.0) at 40 °C for 12 h. Lane Std, mannooligosaccharide standards: mannose (M1), mannobiose (M2), mannotriose (M3), mannotetraose (M4), and mannopentaose (M5).</p>
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6 pages, 880 KiB  
Proceeding Paper
Optimization of Caffeine Extraction from Various Tea Types Using Dichloromethane as an Organic Solvent
by Mariam Mir Memon and Muhammad Idress
Eng. Proc. 2024, 67(1), 81; https://doi.org/10.3390/engproc2024067081 - 18 Dec 2024
Viewed by 312
Abstract
Caffeine, as 1,3,7-trimethyl-1H-purine-2,6-dione, represents a naturally occurring alkaloid within the methyl xanthine family. Exhibiting bitterness and an odorless state in its pure form, caffeine manifests as needle-like crystals. Found abundantly in tea, coffee beans, kola nuts, and cocoa beans, this compound stimulates the [...] Read more.
Caffeine, as 1,3,7-trimethyl-1H-purine-2,6-dione, represents a naturally occurring alkaloid within the methyl xanthine family. Exhibiting bitterness and an odorless state in its pure form, caffeine manifests as needle-like crystals. Found abundantly in tea, coffee beans, kola nuts, and cocoa beans, this compound stimulates the central nervous system, respiration, and cardiac activity. Ubiquitously present in everyday products such as soft drinks, tea, coffee, chocolates, pharmaceutical drugs, and skincare items, caffeine plays a pivotal role. This study focuses on the extraction of caffeine from various tea types, including used tea, utilizing dichloromethane as an organic solvent. Investigating the impact of temperature and residence time on the extraction efficiency, we observed a direct correlation between these factors and efficiency. Notably, the residence time exhibited a significant effect up to a certain threshold, beyond which no substantial difference in extraction efficiency was observed. The optimal conditions for caffeine extraction were identified at 100 °C with a residence time of 30 min. The findings revealed the extraction of 0.089 g of caffeine from black tea, 0.06 g from used black tea, 0.08 g from green tea, and 0.047 g from used green tea. Full article
(This article belongs to the Proceedings of The 3rd International Electronic Conference on Processes)
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<p>Structure of caffeine.</p>
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<p>Extraction of caffeine from different types of teas using dichloromethane.</p>
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<p>Extraction of caffeine from different types of tea using dichloromethane at 80 °C and 100 °C.</p>
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<p>Extraction of caffeine from different types of tea using dichloromethane after 20 min and 30 min.</p>
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16 pages, 1339 KiB  
Article
Chemometric Classification and Bioactivity Correlation of Black Instant Coffee and Coffee Bean Extract by Chlorogenic Acid Profiling
by Yumei Chen, Wei Yu, Yuge Niu, Wenchen Li, Weiying Lu and Liangli (Lucy) Yu
Foods 2024, 13(24), 4016; https://doi.org/10.3390/foods13244016 - 12 Dec 2024
Viewed by 456
Abstract
Chlorogenic acids (CGAs) play a key role in defining the quality and functionality of coffee products. CGA fingerprints of black instant coffee (BIC) and coffee bean extract (CBE) were profiled using ultra-performance liquid chromatography–mass spectrometry and analyzed by chemometrics. A total of 25 [...] Read more.
Chlorogenic acids (CGAs) play a key role in defining the quality and functionality of coffee products. CGA fingerprints of black instant coffee (BIC) and coffee bean extract (CBE) were profiled using ultra-performance liquid chromatography–mass spectrometry and analyzed by chemometrics. A total of 25 CGAs were identified. The BICs yielded higher levels of major CGAs than the CBEs. Furthermore, chemometrics methods, including principal component analysis (PCA) and partial least squares–discriminant analysis (PLS–DA), successfully classified the CBEs and the BICs. In vitro cellular antioxidant activity and viability assays between coffee products further confirmed the relationship between phenolic compounds and bioactivities. Compared to the CBEs, the BICs provided higher cellular toxicity and oxidant activity for hepatocellular carcinoma G2 (HepG2) cells. These results demonstrated that CGAs and their derivatives could be markers for studying coffee-related products. This study revealed the unique phenolic profiles of various coffee products, highlighting the differences between whole beans and soluble coffee. Full article
(This article belongs to the Section Food Analytical Methods)
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<p>The contents of CGAs in BICs and CBEs. * indicates a significant difference in the compound groups (<span class="html-italic">p</span> &lt; 0.01). CQAs and FQAs are the most abundant CGA subgroups in both BICs and CBEs. BICs generally show higher levels of CGAs compared to CBEs, indicating a potential concentration of these compounds during the instant coffee production process. The contents of CQLs, FQLs, and di-CQLs are slightly higher in coffee beans (CBEs) compared to BICs, which may contribute to the bitter taste in whole bean coffees.</p>
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<p>(<b>a</b>) PCA score plots with 1426 variables and (<b>b</b>) with 25 variables; (<b>c</b>) PLS-DA score plots with 1426 variables and (<b>d</b>) with 25 variables. The 95% confidence regions are displayed as color-shadowed ellipses. Both PCA and PLS-DA successfully classify BICs and CBEs, indicating that the phenolic profiles of these two coffee product types are significantly different. The identified CGAs contribute significantly to the classification models, highlighting their role as potential markers for distinguishing between BICs and CBEs. PLS-DA offers better separation compared to PCA, likely due to its supervised learning approach and focus on class separation.</p>
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<p>Effects of CBEs and BICs on (<b>a</b>) cell viability and (<b>b</b>) CAA of HepG2 cells. BICs exhibit significantly lower cell viability compared to CBEs, indicating potential cytotoxic effects; BICs exhibit significantly higher antioxidant activity compared to CBEs.</p>
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<p>Effects of CBEs and BICs on (<b>a</b>) cell viability and (<b>b</b>) CAA of HepG2 cells. BICs exhibit significantly lower cell viability compared to CBEs, indicating potential cytotoxic effects; BICs exhibit significantly higher antioxidant activity compared to CBEs.</p>
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<p>Correlation coefficients of MTT and CAA assays for each individual compound. Most compounds show strong correlations except for compounds 11, 12, 21, 25, etc. MTT and CAA are the complete opposite. For CAA, compounds 1–12, 17, 20, and 25 showed positive correlations.</p>
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17 pages, 1859 KiB  
Article
Genistein and Aphid Probing Behavior: Case Studies on Polyphagous Aphid Species
by Anna Wróblewska-Kurdyk, Bożena Kordan, Katarzyna Stec, Jan Bocianowski and Beata Gabryś
Molecules 2024, 29(23), 5715; https://doi.org/10.3390/molecules29235715 - 3 Dec 2024
Viewed by 542
Abstract
(1) Background: Genistein is a naturally occurring flavonoid with a rich spectrum of biological activities, including plant-herbivore interactions. The aim of the study was to evaluate the effect of exogenous application of genistein on aphid behavior during probing in plant tissues. (2) Methods: [...] Read more.
(1) Background: Genistein is a naturally occurring flavonoid with a rich spectrum of biological activities, including plant-herbivore interactions. The aim of the study was to evaluate the effect of exogenous application of genistein on aphid behavior during probing in plant tissues. (2) Methods: Vicia faba, Brassica rapa ssp. pekinensis, and Avena sativa were treated transepidermally with a 0.1% ethanolic solution of genistein, and the probing behavior of generalist aphid species Aphis fabae, Myzus persicae, and Rhopalosiphum padi was monitored on their respective treated and untreated host plants using electropenetrography (=electrical penetration graph technique, EPG); (3) Results: Genistein did not deter aphid probing activities in non-phloem tissues. In A. fabae and R. padi, a trend towards reduction and in M. persicae a trend towards increase in phloem sap ingestion occurred on genistein-treated plants, but these trends were not statistically significant. (4) Conclusions: Genistein is not a deterrent chemical against generalist aphid species studied; therefore, it is not recommended for practical application. Full article
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<p>Chemical structure of genistein [<a href="#B2-molecules-29-05715" class="html-bibr">2</a>].</p>
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<p>Visualization of aphid stylet activities in plant tissues recorded using electropenetrography (a sample derived from EPG recording of <span class="html-italic">Myzus persicae</span> on <span class="html-italic">Brassica rapa</span> ssp. <span class="html-italic">pekinensis</span> treated with 0.1% genistein). Upper panel illustrates a 1 h section of the 8 h EPG. Lower panels present the details of individual EPG waveforms corresponding with the display in the upper panel. ‘G’—stylets in the xylem (EPG waveform visualizes the active uptake of xylem sap); ‘C’—stylets in epidermis and mesophyll (EPG waveform visualizes the progressive stylet movements within the apoplast and occasional uptake of sap from cells adjacent to the stylet track represented as potential drops ‘pd’); ‘E1’—stylets in phloem (egestion of saliva into sieve elements); ‘E2’—stylets in phloem (passive ingestion of phloem sap from sieve elements).</p>
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<p>Individual variation in probing behavior of aphids on control untreated plants and plants treated transepidermally with 0.1% genistein: (<b>a</b>,<b>b</b>) <span class="html-italic">Aphis fabae</span> on <span class="html-italic">Vicia faba</span>; (<b>c</b>,<b>d</b>) <span class="html-italic">Myzus persicae</span> on <span class="html-italic">Brassica rapa</span> ssp. <span class="html-italic">pekinensis;</span> and (<b>e</b>,<b>f</b>) <span class="html-italic">Rhopalosiphum padi</span> on <span class="html-italic">Avena sativa</span>. Panels (<b>a</b>–<b>f</b>) represent the proportion of time (percentage of cumulative time for individual aphids and the mean of the group) devoted to Np—no probing, C + F + G—pathway + derailed stylet activities + xylem phase, and E—phloem phase E1 (salivation) + E2 (sap ingestion) activities recorded during the 8 h EPG experiments.</p>
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<p>Sequential changes in EPG-recorded stylet penetration activities of aphids on control untreated plants and plants treated transepidermally with 0.1% genistein: (<b>a</b>,<b>b</b>) <span class="html-italic">Aphis fabae</span> on <span class="html-italic">Vicia faba</span>; (<b>c</b>,<b>d</b>) <span class="html-italic">Myzus persicae</span> on <span class="html-italic">Brassica rapa</span> ssp. <span class="html-italic">pekinensis</span>; and (<b>e</b>,<b>f</b>) <span class="html-italic">Rhopalosiphum padi</span> on <span class="html-italic">Avena sativa</span>. Panels (<b>a</b>–<b>f</b>) represent the proportion of time (average percentage of cumulative time for aphids in the group) devoted to Np—no probing, C + F + G—pathway + derailed stylet activities + xylem phase, and E—phloem phase E1 (salivation) + E2 (sap ingestion) activities during the consecutive hours of 8 h EPG recording.</p>
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<p>Sequential changes in EPG-recorded stylet penetration activities of aphids on control untreated plants and plants treated transepidermally with 0.1% genistein: (<b>a</b>,<b>b</b>) <span class="html-italic">Aphis fabae</span> on <span class="html-italic">Vicia faba</span>; (<b>c</b>,<b>d</b>) <span class="html-italic">Myzus persicae</span> on <span class="html-italic">Brassica rapa</span> ssp. <span class="html-italic">pekinensis</span>; and (<b>e</b>,<b>f</b>) <span class="html-italic">Rhopalosiphum padi</span> on <span class="html-italic">Avena sativa</span>. Panels (<b>a</b>–<b>f</b>) represent the proportion of time (average percentage of cumulative time for aphids in the group) devoted to Np—no probing, C + F + G—pathway + derailed stylet activities + xylem phase, and E—phloem phase E1 (salivation) + E2 (sap ingestion) activities during the consecutive hours of 8 h EPG recording.</p>
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<p>Cumulative percentage of aphids that attained phloem phase (=reached sieve elements) during the 8 h EPG monitoring on control untreated plants and plants treated transepidermally with 0.1% genistein: (<b>a</b>,<b>b</b>) <span class="html-italic">Aphis fabae</span> on <span class="html-italic">Vicia faba</span>; (<b>c</b>,<b>d</b>) <span class="html-italic">Myzus persicae</span> on <span class="html-italic">Brassica rapa</span> ssp. <span class="html-italic">pekinensis</span>; and (<b>e</b>,<b>f</b>) <span class="html-italic">Rhopalosiphum padi</span> on <span class="html-italic">Avena sativa</span>. E1—phloem phase salivation represents any contact with sieve elements; E2—phloem phase sap ingestion represents actual feeding, i.e., the uptake of phloem sap.</p>
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<p>Cumulative percentage of aphids that attained phloem phase (=reached sieve elements) during the 8 h EPG monitoring on control untreated plants and plants treated transepidermally with 0.1% genistein: (<b>a</b>,<b>b</b>) <span class="html-italic">Aphis fabae</span> on <span class="html-italic">Vicia faba</span>; (<b>c</b>,<b>d</b>) <span class="html-italic">Myzus persicae</span> on <span class="html-italic">Brassica rapa</span> ssp. <span class="html-italic">pekinensis</span>; and (<b>e</b>,<b>f</b>) <span class="html-italic">Rhopalosiphum padi</span> on <span class="html-italic">Avena sativa</span>. E1—phloem phase salivation represents any contact with sieve elements; E2—phloem phase sap ingestion represents actual feeding, i.e., the uptake of phloem sap.</p>
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26 pages, 3171 KiB  
Article
Producing and Characterizing Polyhydroxyalkanoates from Starch and Chickpea Waste Using Mixed Microbial Cultures in Solid-State Fermentation
by Karlo Grgurević, Dora Bramberger, Martina Miloloža, Krešimir Stublić, Vesna Ocelić Bulatović, Jasmina Ranilović, Šime Ukić, Tomislav Bolanča, Matija Cvetnić, Marinko Markić and Dajana Kučić Grgić
Polymers 2024, 16(23), 3407; https://doi.org/10.3390/polym16233407 - 3 Dec 2024
Viewed by 676
Abstract
The environmental impact of plastic waste is a growing global challenge, primarily due to non-biodegradable plastics from fossil resources that accumulate in ecosystems. Biodegradable polymers like polyhydroxyalkanoates (PHAs) offer a sustainable alternative. PHAs are microbial biopolymers produced by microorganisms using renewable substrates, including [...] Read more.
The environmental impact of plastic waste is a growing global challenge, primarily due to non-biodegradable plastics from fossil resources that accumulate in ecosystems. Biodegradable polymers like polyhydroxyalkanoates (PHAs) offer a sustainable alternative. PHAs are microbial biopolymers produced by microorganisms using renewable substrates, including agro-industrial byproducts, making them eco-friendly and cost-effective. This study focused on the isolation and characterization of PHA-producing microorganisms from agro-industrial waste, including chickpeas, chickpeas with bean residues, and starch. Screening via Sudan Black staining identified PHA-accumulating strains such as Brevibacillus sp., Micrococcus spp., and Candida krusei, among others. To assess the potential for PHA biosynthesis, solid-state fermentation (SSF) was conducted using agro-industrial waste as substrates, along with a mixed culture of the isolated microorganisms. The highest observed yield was a PHA accumulation of 13.81%, achieved with chickpeas containing bean residues. Structural and thermal characterization of the PHAs was performed using Fourier-transform infrared spectroscopy with attenuated total reflectance (FTIR-ATR), differential scanning calorimetry (DSC), and thermogravimetric analysis (TGA). FTIR-ATR spectra indicated polyhydroxybutyrate (PHB), suggesting it as the synthesized PHA type. This study highlights the potential of agro-industrial waste for sustainable PHA production and eco-friendly bioplastics. Full article
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<p>Experimental workflow for PHA production.</p>
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<p>Obtained pure cultures by streaking method, cultures stained with Sudan Black dye, and microphotographs of Gram staining of bacteria isolates <span class="html-italic">Brevibacillus</span> sp. (<b>A</b>,<b>A1</b>,<b>A2</b>), <span class="html-italic">Empedobacter brevis</span>; (<b>B</b>,<b>B1</b>,<b>B2</b>), <span class="html-italic">Aneurinibacillus aneurinilyticus</span>; (<b>C</b>,<b>C1</b>,<b>C2</b>), <span class="html-italic">Micrococcus</span> spp.; (<b>D</b>,<b>D1</b>,<b>D2</b>), <span class="html-italic">Leuconostoc</span> sp; (<b>E</b>,<b>E1</b>,<b>E2</b>), <span class="html-italic">Bacillus licheniformis</span>; (<b>F</b>,<b>F1</b>,<b>F2</b>), <span class="html-italic">Staphylococcus lentus</span>; (<b>G</b>,<b>G1</b>,<b>G2</b>), <span class="html-italic">Citrobacter freundii</span>; (<b>H</b>,<b>H1</b>,<b>H2</b>), M = 1000×.</p>
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<p>Obtained pure cultures by streaking method, cultures stained with Sudan Black dye, and microphotographs of yeast isolates <span class="html-italic">Trichosporon asahii</span> (<b>A</b>,<b>A1</b>,<b>A2</b>), <span class="html-italic">Cryptococcus humicola</span>; (<b>B</b>,<b>B1</b>,<b>B2</b>), <span class="html-italic">Geotrichum klebahnii</span>; (<b>C</b>,<b>C1</b>,<b>C2</b>), <span class="html-italic">Candida krusei</span>; (<b>D</b>,<b>D1</b>,<b>D2</b>), M = 400×.</p>
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<p>FTIR spectra of PHAs obtained in the fermentation processes using (<b>A</b>) chickpea 1; (<b>B</b>) chickpea 2; and (<b>C</b>) starch as a substrate.</p>
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<p>DSC spectra of extracted samples (PHA) obtained in the fermentation processes using (<b>A</b>) chickpea 1; (<b>B</b>) chickpea 2; and (<b>C</b>) starch as a substrate.</p>
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<p>Thermograms of PHAs obtained in the fermentation processes using (<b>A</b>) chickpea 1; (<b>B</b>) chickpea 2; and (<b>C</b>) starch as a substrate.</p>
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18 pages, 6632 KiB  
Article
Efficient and Accurate Calibration of Discrete Element Method Parameters for Black Beans
by Xuezhen Wang, Qinghang Zhai, Shihao Zhang, Qianwen Li and Hanmi Zhou
Agronomy 2024, 14(12), 2803; https://doi.org/10.3390/agronomy14122803 - 25 Nov 2024
Viewed by 389
Abstract
Discrete element parameters of the black bean (BLB) are key to developing high-performance BLB machineries (e.g., seeders and shellers), which are still lacking in previous literature. In this study, the effects of the radius and lifting speed of cylinder-in-cylinder lifting method (CLM) simulations [...] Read more.
Discrete element parameters of the black bean (BLB) are key to developing high-performance BLB machineries (e.g., seeders and shellers), which are still lacking in previous literature. In this study, the effects of the radius and lifting speed of cylinder-in-cylinder lifting method (CLM) simulations were investigated to efficiently and accurately obtain the repose angle. Discrete element method (DEM) parameters of the BLB were determined by combining the Plackett–Burman Design test, the steepest ascent design test, and the central composite design test. The results show that the measurement moment (i.e., 12 s) of repose angles should be determined when kinetic energy reaches the minimal threshold (1 × 10−6 J) to efficiently and accurately obtain repose angles; too early or too late a measurement can result in inaccurate repose angles or excessive computation time of the computer, respectively. The lifting speed and cylinder radius affected the lateral displacements of BLBs and came at the cost of higher computation time and memory usage. A lifting speed of 0.015 m·s−1 and a radius of 40 mm of the cylinder were determined in CLM simulations. The static friction coefficient and rolling friction coefficient between BLBs significantly affected the repose angles. A static friction coefficient of 0.202 and rolling friction coefficient of 0.0104 between BLBs were obtained based on the optimization results. A low relative error (0.74%) and insignificant difference (p > 0.05) between the simulated and measured repose angles were found. The suggested method can be potentially used to calibrate the DEM parameters of BLBs with good accuracy. The results from this study can provide implications for investigating interactions of BLBs and various BLB processing machines and for the efficient and accurate determination of DEM parameters of crop grains. Full article
(This article belongs to the Section Precision and Digital Agriculture)
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<p>The geometrical measurement positions of the black bean (BLB) (<span class="html-italic">L</span> = length; <span class="html-italic">W</span> = width; <span class="html-italic">T</span> = thickness).</p>
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<p>Experimental principle for determining the coefficient of restitution (BLB, H, and h represent black bean, initial height, and the largest rebound height of the black bean, respectively).</p>
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<p>Test principle for measuring the static friction coefficient (BLB and <span class="html-italic">α</span> represent black bean and the angle between the inclined plane and horizontal plane, respectively).</p>
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<p>Test principle for measuring the rolling friction coefficient (x, β, and L represent the distance from the black bean (BLB) to the base edge of the inclined plane, the inclination angle of the plate, and the rolling distance of the BLB on the horizontal plexiglass plate, respectively).</p>
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<p>Cone image from laboratory repose angle test.</p>
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<p>Discrete element modeling of black beans (BLBs): (<b>a</b>) 3D DEM model of a BLB; (<b>b</b>) lifting process model of the cylinder filled with BLBs.</p>
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<p>Dimension distributions of the black bean (BLB): (<b>a</b>) BLB length distribution (i.e., <span class="html-italic">L</span>); (<b>b</b>) BLB width distribution (i.e., <span class="html-italic">W</span>); (<b>c</b>) BLB thickness distribution (i.e., <span class="html-italic">T</span>).</p>
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<p>Laboratory repose angle results of black bean: (<b>a</b>) fit line from the left side; (<b>b</b>) fit line from the right side.</p>
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<p>BLB accumulation forms from lifting speeds of (<b>a</b>) 0.005 m/s, (<b>b</b>) 0.015 m/s, (<b>c</b>) 0.025 m/s, (<b>d</b>) 0.035 m/s, (<b>e</b>) 0.045 m/s, and (<b>f</b>) 0.055 m/s, and (<b>g</b>) effects of the lifting speed on the solution time of the computer and memory usage (i.e., space occupied by simulation data).</p>
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<p>BLB accumulation forms from lifting speeds of (<b>a</b>) 0.005 m/s, (<b>b</b>) 0.015 m/s, (<b>c</b>) 0.025 m/s, (<b>d</b>) 0.035 m/s, (<b>e</b>) 0.045 m/s, and (<b>f</b>) 0.055 m/s, and (<b>g</b>) effects of the lifting speed on the solution time of the computer and memory usage (i.e., space occupied by simulation data).</p>
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<p>The bean particle cones with cylinder radii of (<b>a</b>) 20 mm, (<b>b</b>) 30 mm, (<b>c</b>) 40 mm, (<b>d</b>) 50 mm, and (<b>e</b>) 60 mm, and (<b>f</b>) the effects of the cylinder radius on the number of bean particles and memory usage.</p>
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<p>The bean particle cones with cylinder radii of (<b>a</b>) 20 mm, (<b>b</b>) 30 mm, (<b>c</b>) 40 mm, (<b>d</b>) 50 mm, and (<b>e</b>) 60 mm, and (<b>f</b>) the effects of the cylinder radius on the number of bean particles and memory usage.</p>
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<p>The change in energy during the formation of the repose angle. (<b>a</b>) Particle energy vs. time. (<b>b</b>) Kinetic energy change of the BLB pile after cylinder separation.</p>
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<p>Accumulation angle formation process in releasing stage of bean particles: (<b>a</b>) 4 s; (<b>b</b>) 5 s; (<b>c</b>) 6 s; (<b>d</b>) 7 s; (<b>e</b>) 8 s.</p>
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<p>Change in accumulation angle during 8–15 s.</p>
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<p>Pareto chart.</p>
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<p>Response surface analysis on the effects of static friction and rolling friction coefficients of black beans (BLBs) on the simulated repose angle θx.</p>
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<p>Verification of optimized DEM parameters: (<b>a</b>) repose angle measured in the laboratory. (<b>b</b>) Repose angle from the DEM simulation.</p>
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14 pages, 809 KiB  
Article
Genistein and Naringenin as Defense Molecules
by Sylwia Goławska, Iwona Łukasik and Paweł Czerniewicz
Molecules 2024, 29(23), 5505; https://doi.org/10.3390/molecules29235505 - 21 Nov 2024
Viewed by 435
Abstract
Genistein and naringenin, plant phenolic compounds, are recognized for their health benefits and role in plant defense against herbivores. However, little research exists on how these compounds affect aphid feeding, particularly that of the black bean aphid (Aphis fabae Scopoli) (Hemiptera: Aphididae), [...] Read more.
Genistein and naringenin, plant phenolic compounds, are recognized for their health benefits and role in plant defense against herbivores. However, little research exists on how these compounds affect aphid feeding, particularly that of the black bean aphid (Aphis fabae Scopoli) (Hemiptera: Aphididae), a major pest. This study aimed to evaluate the effects of genistein and naringenin, applied in vitro at different concentrations, on the feeding behavior of A. fabae. Statistical analysis indicated that both the type and concentration of flavonoids significantly influenced aphid stylet activity, salivation, and ingestion. Higher concentrations of both compounds hindered feeding behavior. A longer initial probe was observed on gels containing the studied flavonoids. Genistein at 0.1% completely inhibited salivation while at 0.01%, it reduced the duration of salivation activities. Both compounds also delayed the start and lengthened the duration of active ingestion, though A. fabae tolerated genistein better than naringenin. Naringenin’s effects on feeding behavior were more pronounced at higher concentrations. These findings suggest that genistein and naringenin could be valuable chemicals to protect plants from aphids in a sustainable and environmentally friendly way. Full article
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<p>Chemical structures of naringenin (<b>a</b>) and genistein (<b>b</b>).</p>
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<p>Proportions of time spent by aphids on probing/feeding activities on gels, as affected by naringenin and genistein during 2 h EPG monitoring.</p>
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20 pages, 1941 KiB  
Article
Screening of the Nutritional Properties, Bioactive Components, and Antioxidant Properties in Legumes
by Mihaela Multescu, Alina Culetu and Iulia Elena Susman
Foods 2024, 13(22), 3528; https://doi.org/10.3390/foods13223528 - 5 Nov 2024
Viewed by 2027
Abstract
This study provides an assessment of nutrients (protein, amino acid profiles, fiber, starch), phenolic content TPC, flavonoid content TFC, and antioxidant capacity through different in vitro methods in 12 legume species (red, green, yellow, brown, and black lentils; mung, pinto, black, and kidney [...] Read more.
This study provides an assessment of nutrients (protein, amino acid profiles, fiber, starch), phenolic content TPC, flavonoid content TFC, and antioxidant capacity through different in vitro methods in 12 legume species (red, green, yellow, brown, and black lentils; mung, pinto, black, and kidney beans; chickpea, soy, and lupin) and hemp. Legumes with a protein content above 30% were black lentil, lupin, and soy. Chickpea, soy, black bean, kidney bean, and mung bean did not have any limiting amino acids. All samples had moderate overall protein quality, except green and brown lentils. Black bean was less digestible (68.1%), while soy, hemp, and red lentil had higher protein digestibility (79.3–84.7%). Pinto bean had the highest TPC (425.19 mg GAE/100 g), comparable with hemp, but the lowest TFC (0.24 mg QE/100 g). Yellow and red lentils showed the lowest TPC (69–85.89 mg GAE/100 g). Mung bean presented the highest concentration of flavonoids (45.47 mg QE/100 g), followed by black lentil (28.57 mg QE/100 g). There were distinct variations in the antioxidant capacity across different legume samples and assays. Pinto bean, hemp, and green lentil had the highest relative antioxidant capacity index, while yellow lentil, red lentil, and chickpea presented the lowest. Dark-colored legume samples showed a higher TPC and a lower antioxidant capacity (CUPRAC and PCL assays), while yellow legumes had less antioxidant capacity (DPPH assay). A high correlation coefficient was observed between TPC and DPPH (r = 0.8133), TPC and FRAP (r = 0.8528), TPC and CUPRAC (r = 0.9425), and TPC and ACL (r = 0.8261) methods. The results highlight large variations in the legume properties and support the exploitation of the nutritional properties of legumes as raw materials for the development of products designed to fulfil modern consumer demands. Full article
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<p>In vitro protein digestibility values. Bars with different letters represent a significant difference between samples (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Antioxidant capacity using the Photochem device in an ACL (lipid-soluble antioxidant capacity) system. Bars with different letters represent a significant difference between samples (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Relative antioxidant capacity index in the legumes and hemp samples.</p>
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<p>Correlation between RACI and antioxidant capacity assays: (<b>a</b>) RACI and DPPH; (<b>b</b>) RACI and ABTS; (<b>c</b>) RACI and FRAP; (<b>d</b>) RACI and CUPRAC; (<b>e</b>) RACI and ACL.</p>
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<p>Correlation between RACI and antioxidant capacity assays: (<b>a</b>) RACI and DPPH; (<b>b</b>) RACI and ABTS; (<b>c</b>) RACI and FRAP; (<b>d</b>) RACI and CUPRAC; (<b>e</b>) RACI and ACL.</p>
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<p>Powdered legume colors in the CIE L*a*b* color coordinates are shown as bar length and bar color. The values are presented as mean values ± standard deviation (n = 10). The values followed by different letters indicate significant differences between samples (<span class="html-italic">p</span> &lt; 0.05).</p>
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14 pages, 2851 KiB  
Article
Total Deoxynivalenol Contamination of Wheat Products and Coarse Grains in Shanghai, China: Occurrence and Health Risk Assessment
by Anqi Xu, Shenghao Yu, Yiqi Li, Hong Liu, Zheng Yan, Aibo Wu, Shaojie Peng and Na Liu
Foods 2024, 13(21), 3373; https://doi.org/10.3390/foods13213373 - 23 Oct 2024
Viewed by 1006
Abstract
Deoxynivalenol (DON) is an important mycotoxin produced by Fusarium spp., typically found in cereals, which has garnered considerable research attention. However, the risk assessment of DON exposure to muti-cereal is partial and biased, especially lacking the evaluation of different coarse grains. In this [...] Read more.
Deoxynivalenol (DON) is an important mycotoxin produced by Fusarium spp., typically found in cereals, which has garnered considerable research attention. However, the risk assessment of DON exposure to muti-cereal is partial and biased, especially lacking the evaluation of different coarse grains. In this study, we synthesized and compared the presence of the total deoxynivalenol (free, acetylated, and masked) of a total of 372 grain samples (17 different types) based on high-performance liquid chromatography–tandem mass spectrometry (HPLC–MS/MS), as well as assessed the chronic and acute risks of total DON exposure in the Chinese population. DON was found at the highest frequency with an occurrence of 85.8% (319/372), followed by D3G at 17.2% (64/372). In total, 88.7% (330/372) of the grains were co-contaminated with mycotoxins. The DON and D3G contamination correlation coefficient was 0.68 in wheat flour. Moreover, different DON contamination levels were found in black beans (133.5 µg/kg), soybeans (128.7 µg/kg), and black rice (122.1 µg/kg). The DON/D3G/15A/tDON contamination level was significant differently among different coarse grains. Notably, the Monte Carlo model showed that 3.2–5.9% of adolescents consuming wheat flour and noodles suffered a chronic tDON risk. Full article
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<p>Contamination of detected mycotoxins in total cereals and their products (µg/kg, n = 372) (<b>A</b>) Contamination levels in total samples (the top line of the bar graph represents the mean); The dots means the values of samples. (<b>B</b>) co-occurrence of mycotoxins in total samples; (<b>C</b>) geographical distribution of mycotoxins in total samples (NA: Heilongjiang, Jilin, Liaoning, Inner Mongolia, Beijing, Hebei, and Tianjin; EA: Shandong, Shanghai, Jiangsu, Zhejiang, Anhui, and Fujian; CA: Henan, Hubei, and Hunan; SA: Guangxi, Guangdong, Sichuan, Guizhou, and Yunnan; and Foreign: Japan, Italy, Thailand, Germany, South Korea, and Indonesia).</p>
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<p>Contamination of detected mycotoxins in different cereals and their products. (<b>A</b>) Wheat flour; (<b>B</b>) noodles; (<b>C</b>) fermented products; (<b>D</b>) beer; (<b>E</b>) processed products; and (<b>F</b>) coarse grains. The dots means the values of samples.</p>
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<p>Contamination of detected mycotoxins in 12 types of coarse grains (µg/kg, n = 165). The dots means the values of samples. (<b>A</b>) DON contamination in coarse grains; (<b>B</b>) 3A contamination in coarse grains; (<b>C</b>) 15A contamination in coarse grains; (<b>D</b>) D3G contamination in coarse grains; (<b>E</b>) DONs contamination in coarse grains.</p>
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<p>Probable daily intake estimated from chronic tDON exposure in different cereals and their products for populations performed by deterministic estimation (µg/kg·bw/day). (<b>A</b>) Wheat flour; (<b>B</b>) noodles; (<b>C</b>) fermented products; (<b>D</b>) beer; (<b>E</b>) processed products; (<b>F</b>) coarse grains; and (<b>G</b>) total samples.</p>
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<p>Probable daily intake from acute tDON exposure in different cereals and their products for populations performed by deterministic estimation (µg/kg·bw/day). (<b>A</b>) Wheat flour; (<b>B</b>) noodles; (<b>C</b>) fermented products; (<b>D</b>) beer; (<b>E</b>) processed products; (<b>F</b>) coarse grains; and (<b>G</b>) total samples. The dashed line means the ARfD value.</p>
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<p>Probable daily intake intake from tDON exposure in different cereals and their products for populations performed by probabilistic estimation (µg/kg·bw/day). (<b>A</b>) Wheat flour; (<b>B</b>) noodles; (<b>C</b>) fermented products; (<b>D</b>) beer; (<b>E</b>) processed products; (<b>F</b>) coarse grains; and (<b>G</b>) total samples.</p>
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12 pages, 2548 KiB  
Article
Morpho–Molecular Characterization of Brazilian Common Bean Landraces Varieties and Commercial Cultivars
by Ana Claudia Schllemer dos Santos, Isadora Bischoff Nunes, Lucas Teixeira da Silva, Lucas Vinícius de Sousa Alcântara, Lucas da Silva Domingues, Joel Donazzolo, Juliana Morini Kupper Cardoso Perseguini and Jean Carlo Possenti
Horticulturae 2024, 10(11), 1123; https://doi.org/10.3390/horticulturae10111123 - 22 Oct 2024
Viewed by 676
Abstract
This study aimed to characterize forty genotypes (31 landrace varieties and nine commercial cultivars) of common bean (Phaseolus vulgaris L.) morphologically and molecularly. Morphological descriptors were evaluated during seedling, flowering, physiological maturation and post-harvest stages. Ten microsatellite markers were used for molecular [...] Read more.
This study aimed to characterize forty genotypes (31 landrace varieties and nine commercial cultivars) of common bean (Phaseolus vulgaris L.) morphologically and molecularly. Morphological descriptors were evaluated during seedling, flowering, physiological maturation and post-harvest stages. Ten microsatellite markers were used for molecular screening. The markers were analyzed according to the number of alleles per locus, the allele frequency per locus and the polymorphism in content (PIC). Genetic distances and cluster analysis were performed using Bayesian inference and the UPGMA method. All black beans evaluated have anthocyanin in the cotyledons, hypocotyls and stems, and their flowers are purple. For the growth habit, 50% of the evaluated genotypes belong to type III, and at the physiological maturation stage, yellow color is predominant in the pods (85%). Through the genetic dissimilarity, three groups were observed for the likelihood reason, and five groups were observed through the UPGMA method, a strong indication of the wide genetic diversity among the evaluated genotypes. All genotypes from the Andean center of origin were grouped into the same cluster. Full article
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<p>Morphological diversity of seeds of the 31 landraces (1 to 31) and the 9 commercial cultivars (32 to 40) used in this study.</p>
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<p>Representation of the 40 common bean genotypes according to Bayesian inference. The evaluated accessions were divided into 3 subpopulations (K = 3), represented by the gray bar on top. The accesses are represented by the colored bars. Different accesses with the same color belong to the same subpopulation. Different colors in the same access indicate the percentage of genome shared between each subpopulation.</p>
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<p>Unrooted dendrogram of the 40 common bean genotypes through the UPGMA method based on morphological and molecular data obtained in this study.</p>
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18 pages, 1663 KiB  
Article
Co-Encapsulation of Coffee and Coffee By-Product Extracts with Probiotic Kluyveromyces lactis
by Dérica Gonçalves Tavares, Mayara Andrade Martins de Souza, Tamara Leite dos Santos, Adriele do Amor Divino Silva, Danilo José Machado de Abreu and Whasley Ferreira Duarte
Foods 2024, 13(19), 3056; https://doi.org/10.3390/foods13193056 - 26 Sep 2024
Viewed by 1200
Abstract
Coffee and coffee by-products contain several chemical compounds of great relevance, such as chlorogenic acid (CGA), trigonelline, and caffeine. Furthermore, yeasts have been the target of studies for their use as probiotics because of their interesting biochemical characteristics. The combined administration of probiotic [...] Read more.
Coffee and coffee by-products contain several chemical compounds of great relevance, such as chlorogenic acid (CGA), trigonelline, and caffeine. Furthermore, yeasts have been the target of studies for their use as probiotics because of their interesting biochemical characteristics. The combined administration of probiotic microorganisms with components that provide health benefits mediated by alginate encapsulation is an alternative that ensures the stability of cells and chemical compounds. In this context, the aim of this work was to co-encapsulate the probiotic yeast Kluyveromyces lactis B10 and extracts of green coffee beans, coffee silverskin, and PVA (black, green or immature, and sour coffee beans). The bioactive composition, antioxidant and antimicrobial activities of the extracts, microcapsule morphological characteristics and encapsulation efficiency, ability of the encapsulation to protect the yeast cells subjected to gastrointestinal conditions, and antioxidant activity of the microcapsules were evaluated. All the evaluated extracts showed antioxidant activity, of which PVA showed 75.7% and 77.0%, green coffee bean showed 66.4% and 45.7%, and coffee silverskin showed 67.7% and 37.4% inhibition of DPPH and ABTS•+ radicals, respectively, and antimicrobial activity against the pathogenic bacteria E. coli, Salmonella, and S. aureus, with high activity for the PVA extract. The microcapsules presented diameters of between 1451.46 and 1581.12 μm. The encapsulation efficiencies referring to the yeast retention in the microcapsules were 98.05%, 96.51%, and 96.32% for green coffee bean, coffee silverskin, and PVA, respectively. Scanning electron microscopy (SEM) showed that the microcapsules of the three extracts presented small deformations and irregularities on the surface. The K. lactis cells encapsulated in all treatments with the extracts showed viability higher than 8.59 log CFU/mL, as recommended for probiotic food products. The addition of green coffee bean, coffee silverskin, and PVA extracts did not reduce the encapsulation efficiency of the alginate microcapsules, enabling a safe interaction between the extracts and the K. lactis cells. Full article
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<p>Antioxidant activity of green coffee bean, coffee silverskin, and PVA extracts by the 1-diphenyl-2-picrylhydrazyl (DPPH) and 2′-azino-bis-3-ethylbenzthiazoline-6-sulfonic acid (ABTS<sup>•+</sup>) methods. Data are expressed as triplicate mean ± standard deviation. Means with different letters are significantly different at <span class="html-italic">p</span> &lt; 0.05 by Scott–Knott test.</p>
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<p>SEM and fluorescence microscopy of microcapsules. (<b>a</b>) External structure of the microcapsule; (<b>b</b>) distribution of <span class="html-italic">K. lactis</span> B10 in the microcapsule in groups of cells (arrow); (<b>c</b>) detail of the alginate porous network; (<b>d</b>) microcapsule with green coffee bean extract showing <span class="html-italic">K. lactis</span> B10 group of cells and cells that have undergone budding (arrows); (<b>e</b>) microcapsule with coffee silverskin extract showing <span class="html-italic">K. lactis</span> B10 group of cells; (<b>f</b>) microcapsule with PVA extract showing <span class="html-italic">K. lactis</span> B10 group of cells; (<b>g</b>–<b>i</b>) viability of <span class="html-italic">K. lactis</span> B10 encapsulated by the live–dead test. Dead cells were stained with Propidium Iodide (red) and living cells were stained with Syto9 (green). Microcapsules with green coffee bean extract (<b>g</b>), PVA extract (<b>h</b>), and coffee silverskin extract (<b>i</b>).</p>
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17 pages, 16970 KiB  
Article
Effects of Media and Processes on the Aromas of White Truffle Solid-State Fermented Products
by Chih-Yuan Cheng and Su-Der Chen
Processes 2024, 12(9), 2036; https://doi.org/10.3390/pr12092036 - 21 Sep 2024
Viewed by 626
Abstract
This study aimed to formulate a black bean soy sauce using black beans and black rice as media for the solid-state fermentation of white truffle. Various proportions of these media (4:0, 3:1, 2:2, 1:3, and 0:4) were prepared, with methionine concentrations (0, 0.3, [...] Read more.
This study aimed to formulate a black bean soy sauce using black beans and black rice as media for the solid-state fermentation of white truffle. Various proportions of these media (4:0, 3:1, 2:2, 1:3, and 0:4) were prepared, with methionine concentrations (0, 0.3, 0.6, 0.9, 1.2, and 1.5%) serving as precursors for a 4-week solid-state fermentation to analyze the aroma profiles. GC-MS analysis showed that samples with 1.5% methionine exhibited significantly higher levels of sulfur-containing volatile compounds compared to those without methionine. GC-IMS analysis revealed that a 2:2 ratio of black beans to black rice produced the most enriched aroma. Lower methionine levels improved mycelial growth, with 0.3% methionine yielding the richest aroma components. After fermentation, the white truffle products were sterilized using autoclaving, hot air assisted radio frequency (HARF), and high pressure processing (HPP), followed by freeze drying. GC-IMS analysis showed that HPP samples had an aroma closest to fresh samples, whereas HARF and autoclave resulted in similar aromas. However, 24 h freeze drying significantly diminished the aroma, resulting in no significant difference in aroma among the freeze-dried products treated with different sterilization methods. Full article
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<p>Experimental design of aroma study in <span class="html-italic">Tuber magnatum</span> solid-state fermentation.</p>
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<p>GC-MS spectrum of <span class="html-italic">Tuber magnatum</span> four-week solid-state fermented products using different ratios of black bean and black rice as media.</p>
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<p>GC-MS spectrum of <span class="html-italic">Tuber magnatum</span> four-week solid-state fermented products using different ratios of black bean and black rice as media with 1.5% methionine.</p>
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<p>Aroma fingerprints of <span class="html-italic">Tuber magnatum</span> solid-state fermented products using different ratios of black bean and black rice as media.</p>
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<p>The characteristic aroma fingerprints of <span class="html-italic">Tuber magnatum</span> solid-state fermented products using different ratios of black bean and black rice as media.</p>
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<p>Principal component analysis of <span class="html-italic">Tuber magnatum</span> solid-state fermented products using different ratios of black bean and black rice as media.</p>
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<p>Aroma fingerprints of <span class="html-italic">Tuber magnatum</span> solid-state fermented products using different concentrations of methionine in media.</p>
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<p>The characteristic aroma fingerprints of <span class="html-italic">Tuber magnatum</span> solid-state fermented products with different concentrations of methionine in media.</p>
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<p>Aroma fingerprints of unfermented products with different concentrations of methionine in media.</p>
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<p>The characteristic aroma fingerprints of (<b>a</b>) before and (<b>b</b>) after <span class="html-italic">Tuber magnatum</span> solid-state fermented products with different concentrations of methionine in media.</p>
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<p>Principal component analysis of (<b>a</b>) before and (<b>b</b>) after <span class="html-italic">Tuber magnatum</span> solid-state fermented products with different concentrations of methionine in media.</p>
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<p>Aroma fingerprints of different pasteurization methods and their freeze-dried (<b>a</b>) before and (<b>b</b>) after <span class="html-italic">Tuber magnatum</span> solid-state fermented products.</p>
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<p>Aroma fingerprints of different pasteurization methods and their freeze-dried (<b>a</b>) before and (<b>b</b>) after <span class="html-italic">Tuber magnatum</span> solid-state fermented products.</p>
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<p>(<b>a</b>) Before and (<b>b</b>) after characteristic aroma compound fingerprints of <span class="html-italic">Tuber magnatum</span> solid-state fermented products by different pasteurization methods and freeze-drying.</p>
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<p>(<b>a</b>) Before and (<b>b</b>) after characteristic aroma compound fingerprints of <span class="html-italic">Tuber magnatum</span> solid-state fermented products by different pasteurization methods and freeze-drying.</p>
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<p>Principal component analysis of <span class="html-italic">Tuber magnatum</span> solid-state fermented products using different pasteurization methods and freeze-drying.</p>
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30 pages, 1445 KiB  
Review
An Overview of Aspergillus Species Associated with Plant Diseases
by Latiffah Zakaria
Pathogens 2024, 13(9), 813; https://doi.org/10.3390/pathogens13090813 - 20 Sep 2024
Cited by 2 | Viewed by 3026
Abstract
The genus Aspergillus contains several species that are important plant pathogens. Plant pathogenic Aspergillus spp. affect agricultural crops in the field as well as after harvest, often associated with corn ear rot, cotton boll rot, peanut yellow mold, black mold of onion and [...] Read more.
The genus Aspergillus contains several species that are important plant pathogens. Plant pathogenic Aspergillus spp. affect agricultural crops in the field as well as after harvest, often associated with corn ear rot, cotton boll rot, peanut yellow mold, black mold of onion and garlic, fruit rot on grapes, pomegranates, olives, citrus, and apples. Coffee berries and coffee beans as well as tree nuts are also frequently infected by Aspergillus spp. Some of the plant pathogenic Aspergillus spp. are also mycotoxigenic, produced mycotoxin in the plant tissues leading to contamination of agricultural products. Over the years, reports of plant diseases caused by Aspergillus in various crops have increased, suggesting they are commonly encountered plant pathogens. This review focuses on agricultural crops or cultivated plants infected by Aspergillus spp. The compilation of plant pathogenic Aspergillus spp. provides information to mycologists, particularly those involved in plant pathology and crop protection, with updated information on plant diseases caused by various species of Aspergillus. The updated information also includes the locality or location, province, state and the country. The knowledge on the prevalence and geographic distribution of plant pathogenic Aspergillus spp. is beneficial in the application of crop protection. Full article
(This article belongs to the Special Issue Filamentous Fungal Pathogens: 2nd Edition)
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<p>Black mold of onion (<b>A</b>) and garlic (<b>B</b>).</p>
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<p>Infection of <span class="html-italic">Aspergillus</span> spp. on various fruit crops in the field and after harvest.</p>
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15 pages, 1948 KiB  
Article
Nutritional Qualities, Metabolite Contents, and Antioxidant Capacities of Yardlong Beans (Vigna unguiculata subsp. sesquipedalis) of Different Pod and Seed Colors
by Yu-Mi Choi, Myoung-Jae Shin, Hyemyeong Yoon, Sukyeung Lee, Jungyoon Yi, Xiaohan Wang and Kebede Taye Desta
Antioxidants 2024, 13(9), 1134; https://doi.org/10.3390/antiox13091134 - 19 Sep 2024
Viewed by 1094
Abstract
Studying the effects of genetic and environmental factors on plant biochemical components helps in selecting the best varieties for the food industry and breeding programs. This study analyzed the nutritional qualities, secondary metabolites, and antioxidant activities of 14 field-grown yardlong beans accessions and [...] Read more.
Studying the effects of genetic and environmental factors on plant biochemical components helps in selecting the best varieties for the food industry and breeding programs. This study analyzed the nutritional qualities, secondary metabolites, and antioxidant activities of 14 field-grown yardlong beans accessions and how they are affected by differences in pod and seed colors. The analyzed parameters varied significantly among the yardlong bean accessions, with variances ranging from 1.36% in total unsaturated fatty acid content to 51.01% in DPPH scavenging activity. Accessions YLB4, YLB7, and YLB14 performed the best, showing antioxidant indices of 100.00, 70.10, and 67.88%, respectively. Among these, YLB14 showed a characteristic property, having the highest levels of vitamin C (2.62 mg/g) and omega-6 to omega-3 ratio (2.67). It also had the second highest dietary fiber (21.45%), stearic acid (4.44%), and linoleic acid (40.39%) contents, as well as the lowest thrombogenicity index (0.38). Although cluster and principal component analyses did not clearly separate the yardlong beans based on pod or seed color, analysis of variance revealed that these factors and their interaction had significant effects on total phenol, DPPH scavenging activity, ABTS•+ scavenging activity, and reducing power. In contrast, the nutritional parameters, except for dietary fiber, were not significantly affected by pod and seed color variations. Therefore, consuming yardlong beans of different pod and seed colors may not affect the overall nutrient intake. In general, this study identified yardlong beans with green pods and black seeds as good sources of antioxidants. Accordingly, further metabolomics and genomics studies are suggested to thoroughly explore their characteristics. Full article
(This article belongs to the Special Issue Plant Materials and Their Antioxidant Potential, 2nd Edition)
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<p>The contents of crude protein (<b>a</b>), total fat (<b>b</b>), crude fiber (<b>c</b>), and dietary fiber (<b>d</b>) across 14 yardlong bean accessions of different immature pod colors (green and light green) and seed colors (brown and black). Different letters on bars (charts) indicate significantly different means at <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>LC-chromatogram of vitamin C in representative samples with light green pod and black seed color (<b>a</b>) and green pod and brown seed color (<b>b</b>), and the content of vitamin C in 14 yardlong bean accessions (<b>c</b>). Peak assignment: 1: Vitamin C. Different letters on bars indicate significantly different mean values at <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Dendrogram of 14 yardlong bean accessions (<b>a</b>), score plot of yardlong bean accessions according to seed color (<b>b</b>) and loading plot of variable (<b>c</b>) from PCA. ABTS: ABTS<sup>•+</sup> scavenging activity, CFC: crude fiber content, DFC: dietary fiber content, DPPH: DPPH<sup>•</sup> scavenging activity, LA: linoleic acid, LLA: linolenic acid, OA: oleic acid, PA: palmitic acid, PUAF: total polyunsaturated fatty acid, RP: reducing power, SA: stearic acid, TF: total fat, TP: total protein, TSFA: total saturated fatty acid, TUFA: total unsaturated fatty acid, TPC: total phenolic content, TSC: total saponin content, TTC: total tannin content, Vit C: vitamin C.</p>
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<p>Pearson’s correlation matrix showing the association between the analyzed quantitative parameters. *** <span class="html-italic">p</span> &lt; 0.001, ** <span class="html-italic">p</span> &lt; 0.01, * <span class="html-italic">p</span> &lt; 0.05, ns: not significant. ABTS: ABTS<sup>•+</sup> scavenging activity, CFC: crude fiber content, DFC: dietary fiber content, DPPH: DPPH<sup>•</sup> scavenging activity, LA: linoleic acid, LLA: linolenic acid, OA: oleic acid, PA: palmitic acid, PUFA: total polyunsaturated fatty acid, RP: reducing power, SA: stearic acid, TF: total fat, TP: total protein, TSFA: total saturated fatty acid, TUFA: total unsaturated fatty acid, TPC: total phenolic content, TSC: total saponin content, TTC: total tannin content, Vit C: vitamin C.</p>
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