Levels Of Proanthocyanidins Research Articles

Temporal changes in the proanthocyanidins to anthocyanins ratio during dormancy associate with bloom time variations in peach

Our previous research demonstrated that fall applications of ethephon, an ethylene-releasing plant growth regulator, delay bloom in peach, accompanied by changes in endogenous hormones, ROS, sugar metabolism, and transcriptomic profiles during bud dormancy phases (endodormancy and ecodormancy). In this study, floral bud tissues were collected from ethephon-treated and untreated trees at three time points (200, 600, and 1000 chilling hours, CH) during endodormancy and two points (1000 and 3000 growing degree hours, GDH) during ecodormancy. Using ultra-performance liquid chromatography time-of-flight mass spectrometry (UPLC-TOF/MS), we aimed to unravel the untargeted metabolic changes explaining ethephon-mediated bloom delay. Metabolite set-enrichment analysis (MSEA) revealed significant chemical group variations between dormancy phases, with a threefold increase in flavonoids during endodormancy and a doubling of organic and amino acids during ecodormancy. Further analysis of genes associated with the biosynthesis and transcriptional regulation of the flavonoid pathway showed that ethephon treatment upregulated genes associated with proanthocyanidin (PA) biosynthesis and downregulated genes related to anthocyanins (ACNs). We quantified PA and ACN contents in 12 peach cultivars with contrasting bloom times and chilling requirements (e.g. 727–1308 CH). Late-bloom cultivars had higher PA levels during endodormancy, while early-bloom cultivars had higher ACN levels during ecodormancy. Staining buds with 4-dimethylaminocinnamaldehyde (DMAC) dye revealed a decline in the PA/ACN ratio at later ecodormancy stages, correlating with bloom time. Integrated analysis of metabolite content and gene expression in late-bloom 'KV021779' and early-bloom 'John Boy' cultivars validated that late-blooming cultivars have higher PA levels during endodormancy, extending dormancy-release periods and resulting in later blooms.

Targeted regulation of 5-aminolevulinic acid enhances flavonoids, anthocyanins and proanthocyanidins accumulation in Vitis davidii callus

BackgroundSpine grape (Vitis davidii) is a promising source of high-quality anthocyanins, with vast potential for application in food, pharmaceutical, and cosmetic industries. However, their availability is limited by resource constraints. Plant cell culture has emerged as a valuable approach for anthocyanin production and serves as an ideal model to investigate the regulation of anthocyanin biosynthesis. Elicitors are employed to achieve targeted enhancement of anthocyanin biosynthesis. The present study investigated the impact of 5-aminolevulinic acid (ALA) as an elicitor on the accumulation of anthocyanins and flavonoids during spine grape callus growth. Specifically, we examined the effects of ALA on anthocyanin and its component accumulation in callus, and biosynthetic anthocyanin gene expression.ResultsALA at 25 µg/L increased the biomass of spine grape callus. ALA induction enhanced the levels of flavonoids, anthocyanins and proanthocyanidins in callus, with maximum values reaching 911.11 mg/100 g DW, 604.60 mg/100 g DW, and 5357.00 mg/100 g DW, respectively, after callus culture for 45 days. Notably, those levels were 1.47-, 1.93- and 1.83-fold higher than controls. ALA induction modulated the flavonoid profile, and among 97 differential flavonoid metabolites differing from controls, 77 were upregulated and 20 were downregulated. Six kinds of anthocyanins, namely cyanidin (8), delphinidin (6), peonidin (5), malvidin (4), petunidin (3) and pelargonidin (3), were detected in callus, with peonidin most abundant. Compared with controls, anthocyanin components were increased in ALA-treated callus. The key genes PAL1, PAL2, PAL4, CHI, CHS3, F3’H, F3H, FLS, DFR, UFGT, MYBA1, LDOX, OMT3, GT1 and ACT involved in anthocyanin biosynthesis were upregulated following ALA treatment, resulting in anthocyanin accumulation.ConclusionThis study revealed a novel mode of ALA-mediated promotion of plant anthocyanin biosynthesis and accumulation at the cellular level, and a strategy for enhancing anthocyanin content in spine grape callus. The findings advance commercial-scale production of anthocyanins via spine grape callus culture. we also explored the accumulation patterns of flavonoids and anthocyanins under ALA treatment. Augmentation of anthocyanins coincided with elevated expression levels of most genes involved in anthocyanin biosynthesis within spine grape callus following ALA treatment.

Loss-of-function mutation in anthocyanidin reductase activates the anthocyanin synthesis pathway in strawberry

Fruit color substantially affects consumer preferences, with darker red strawberries being economically more valuable due to their higher anthocyanin content. However, the molecular basis for the dark red coloration remains unclear. Through screening of an ethyl methanesulfonate mutant library, we identified a rg418 mutant, that demonstrated anthocyanin accumulation during early fruit development stages. Furthermore, the ripening fruits of this mutant had higher anthocyanin content than wild-type (WT) fruits. An analysis of flavonoid content in WT and rg418 mutant fruits revealed substantial changes in metabolic fluxes, with the mutant exhibiting increased levels of anthocyanins and flavonols and decreased levels of proanthocyanidins. Bulked sergeant analysis sequencing indicated that the mutant gene was anthocyanidin reductase (ANR), a key gene in the proanthocyanidin synthesis pathway. Furthermore, transcriptome sequencing revealed the increased expression of MYB105 during the early development stage of mutant fruits, which promoted the expression of UFGT (UDP-glucose flavonoid 3-O-glucosyltransferase), a key gene involved in anthocyanin synthesis, thus substantially enhancing the anthocyanin content in the mutant fruits. Additionally, mutating ANR in a white-fruited strawberry variant (myb10 mutant) resulted in appealing pink-colored fruits, suggesting the diverse roles of ANR in fruit color regulation. Our study provides valuable theoretical insights for improving strawberry fruit color.

Antioxidant and Antimicrobial Properties of Stachys maritima via Quantum Dots and Molecular Docking.

Stachys maritima is a species of plant belonging to the Lamiaceae, commonly known as common sea lavender. The extraction of S. maritima involves drying the aerial parts, grinding them into powder, followed by extraction with methanol. ST-CQDs (S. maritima carbon quantum dots) were synthesized from S. maritima extract via pyrolysis. An optimal synthesis condition of 7 hours of pyrolysis was determined to yield ST-CQDs with high absorption in the visible-UV range. Characterization techniques such as UV-visible spectroscopy and TEM (Transmission Electron Microscopy) confirmed the spherical and homogeneous nature of the carbon quantum dots. S. maritima methanol extract exhibited strong antioxidant activity with a DPPH (2,2-Diphenyl-1-picrylhydrazyl) IC50 value of 0.114±0.001 mg/mL. Similarly, ST-CQDs showed strong antioxidant properties with a DPPH IC50 value of 0.69±0.03 mg/mL. Moreover, the methanol extract of S. maritima demonstrated antimicrobial activity against E. coli ATCC 25922 and S. aureus ATCC 25923, with effective MIC values of 25 mg/mL and 6.25 mg/mL, respectively. However, ST-CQDs did not show antimicrobial effects against the tested microorganisms (E. coli ATCC 25922, S. aureus ATCC 25923, K. pneumoniae ATCC 13883, and C. albicans ATCC 10231). Molecular docking simulations suggested that compounds derived from S. maritima (such as 9,12,15-octadecatrienoic acid and palmitic acid) could interact effectively with TNF-α (Tumor Necrosis Factor Alpha), indicating potential anti-inflammatory properties. This study highlights that the methanol extract of S. maritima possesses potent antioxidant and antimicrobial activities, and ST-CQDs exhibit similar antioxidant properties. However, ST-CQDs did not demonstrate antimicrobial activity against the tested pathogens. Molecular docking simulations also suggest potential anti-inflammatory properties of compounds derived from the plant.

Antioxidant, anti-tyrosinase, hepatoprotective, and anti-inflammatory potential in flowers and seeds of Ochna integerrima (Lour.) Merr

This study explored, for the first time, the antioxidant (total antioxidant content, reducing power, ferric ion reducing antioxidant power, hydroxyl radical scavenging, ferrous ion-chelating assays), anti-tyrosinase, anti-inflammatory properties, and hepatoprotective effect in HepG2 cell lines of Ochna integerrima (Loureiro) Merrill flowers and seeds. All extracts except n-hexane exhibited significant antioxidant activity, with high levels of tannin and proanthocyanidins. Luteolin (1), 6-γ,γ-dimethylallylkaempferol7-O-β-d-glucopyranoside (2), 6-γ,γ-dimethylallylquercetin7-O-β-d-glucopyranoside (3), and 6-γ,γ-dimethylallyldihydrokaempferol 7-O-β-d-glucopyranoside (4) were isolated using semi-preparative HPLC. Compounds 1–3 demonstrated good anti-tyrosinase activity. The most active hepatoprotective extracts were found to be aqueous extracts. The flower extracts exhibited greater anti-inflammatory properties by the decrease of NO in RAW 264.7 cells and bovine serum albumin protein. Among them, the n-hexane and EtOAc extracts from flowers displayed promising anti-inflammatory activity. This was predicted by in silico analysis of 1–4. In summary, O. integerrima appears to be a promising natural source for antioxidant, anti-tyrosinase, and anti-inflammatory applications.

Exogenous application of sucrose promotes the repartitioning of anthocyanin and proanthocyanidin in ‘Cabernet Sauvignon’ grapevine berries

Sugar and its metabolism play a critical role in fruit quality and determine the accumulation characteristics of secondary metabolites. In this study, the dynamic changes in anthocyanin and proanthocyanidin (PA) accumulation during exogenous sugar application were investigated throughout the grape ripening stage, and the underlying biosynthesis mechanism was monitored. Results showed that exogenous sucrose and N-acetyl-d-glucosamine (hexokinase inhibitor, HI) promoted grape berry ripening. Exogenous application of sucrose and HI significantly increased the levels of soluble sugars and anthocyanins, while significantly decreased the levels of acids and PAs in grape berries. Exogenous sucrose and HI activated the transcription of the VvSPS, VvAI and VvNI genes but inhibited the transcription of VvHXK gene to alter sugar metabolism. Notably, the accumulation of anthocyanin compounds, particularly peonidin derivatives, significantly increased in the sucrose- and HI-treated berries by upregulating the transcription of anthocyanin biosynthesis genes, including VvCHS2, VvF3′H, VvDFR, VvLDOX, and VvUFGT, whereas the accumulation of PA units, such as (‒)-epicatechin, (+)-gallocatechin, and (‒)-epigallocatechin, was repressed by downregulating the transcription of the VvLAR2 and VvANR genes, which boosted anthocyanin and PA reprogramming. These results provide preliminary information on how to modulate the biosynthesis and accumulation of anthocyanins and PAs in grape berries by the exogenous application of sucrose and thus, to modify berry composition.

Variation in Nutritional Components and Antioxidant Capacity of Different Cultivars and Organs of Basella alba

Basella alba is a frequently consumed leafy vegetable. However, research on its nutritional components is limited. This study aimed to explore the variation in the nutritional components and antioxidant capacity of different cultivars and organs of Basella alba. Here, we primarily chose classical spectrophotometry and high-performance liquid chromatography (HPLC) to characterize the variation in nutritional components and antioxidant capacity among different organs (inflorescences, green fruits, black fruits, leaves, and stems) of eight typical cultivars of Basella alba. The determination indices (and methods) included the total soluble sugar (anthrone colorimetry), total soluble protein (the Bradford method), total chlorophyll (the ethanol-extracting method), total carotenoids (the ethanol-extracting method), total ascorbic acid (the HPLC method), total proanthocyanidins (the p-dimethylaminocinnamaldehyde method), total flavonoids (AlCl3 colorimetry), total phenolics (the Folin method), and antioxidant capacity (the FRAP and ABTS methods). The results indicated that M5 and M6 exhibited advantages in their nutrient contents and antioxidant capacities. Additionally, the inflorescences demonstrated the highest total ascorbic acid and total phenolic contents, while the green and black fruits exhibited relatively high levels of total proanthocyanidins and antioxidant capacity. In a comparison between the green and black fruits, the green fruits showed higher levels of total chlorophyll (0.77–1.85 mg g−1 DW), total proanthocyanidins (0.62–2.34 mg g−1 DW), total phenolics (15.28–27.35 mg g−1 DW), and ABTS (43.39–59.16%), while the black fruits exhibited higher levels of total soluble protein (65.45–89.48 mg g−1 DW) and total soluble sugar (56.40–207.62 mg g−1 DW) in most cultivars. Chlorophyll, carotenoids, and flavonoids were predominantly found in the leaves of most cultivars, whereas the total soluble sugar contents were highest in the stems of most cultivars. Overall, our findings underscore the significant influence of the cultivars on the nutritional composition of Basella alba. Moreover, we observed notable variations in the nutrient contents among the different organs of the eight cultivars, and proanthocyanidins may contribute significantly to the antioxidant activity of the fruits. On the whole, this study provides a theoretical basis for the genetic breeding of Basella alba and dietary nutrition and serves as a reference for the comprehensive utilization of this vegetable.

Multiple bHLH/MYB-based protein complexes regulate proanthocyanidin biosynthesis in the herbage of Lotus spp.

Main conclusionThe complexes involving MYBPA2, TT2b, and TT8 proteins are the critical regulators of ANR and LAR genes to promote the biosynthesis of proanthocyanidins in the leaves of Lotus spp.The environmental impact and health of ruminants fed with forage legumes depend on the herbage's concentration and structure of proanthocyanidins (PAs). Unfortunately, the primary forage legumes (alfalfa and clover) do not contain substantial levels of PAs. No significant progress has been made to induce PAs to agronomically valuable levels in their edible organs by biotechnological approaches thus far. Building this trait requires a profound knowledge of PA regulators and their interplay in species naturally committed to accumulating these metabolites in the target organs. Against this background, we compared the shoot transcriptomes of two inter-fertile Lotus species, namely Lotus tenuis and Lotus corniculatus, polymorphic for this trait, to search for differentially expressed MYB and bHLH genes. We then tested the expression of the above-reported regulators in L. tenuis x L. corniculatus interspecific hybrids, several Lotus spp., and different L. corniculatus organs with contrasting PA levels. We identified a novel MYB activator and MYB-bHLH-based complexes that, when expressed in Nicotiana benthamiana, trans-activated the promoters of L. corniculatus anthocyanidin reductase and leucoanthocyanidin reductase 1 genes. The last are the two critical structural genes for the biosynthesis of PAs in Lotus spp. Competition between MYB activators for the transactivation of these promoters also emerged. Overall, by employing Lotus as a model genus, we refined the transcriptional network underlying PA biosynthesis in the herbage of legumes. These findings are crucial to engineering this trait in pasture legumes.

Effect of Adding Different Levels of Proanthocyanidin in Laying Hens Diet on Productive Performance

The research was carried out at the college of Agriculture/Al-Qasim Green University’s Department of Animal Production farm. The experimental field trial ran for 16 weeks, from March 19, 2022, to July 2, 2022 (divided into four 4-week phases). The purpose of this study was to determine the impact of supplementing chicken diets with varying concentrations of the Proanthocyanidin chemical on egg production, quality, and oxidation markers. During the experiment’s breeding phase, which was split into four experimental periods (ages 53–56 weeks, 57–60 weeks, 61–64 weeks, and 65–68 weeks), 105 Lohmann brown laying hens were used.15 pens were set up with 5 experimental treatments for each treatment of 21 birds, and each treatment contained 3 duplicates per 7 birds, all fed according to the standards outlined in the Lohmann Brown Layer Managements Guide. The following procedures were used in the pilot programme:In the first phase, a non-experimental group served as a control. Second, 100 milligrammes of proanthocyanidin per kilogramme of feed was given to a standard diet. Third, we have a standard diet with 200 milligrammes of proanthocyanidin per kilogramme of feed. Fourthly, a standard diet with 300 milligrammes of Proanthocyanidin per kilogramme of feed was used. Fifthly, we have a standard diet with 400 milligrammes of proanthocyanidin per kilogramme of feed. Key findings from the research include: Compared to the control group, the third, fourth, and fifth treatments significantly increased (p0.05) egg production rate, cumulative number of eggs, egg weight, egg mass, and feed conversion coefficient. - Shell thickness (in millimetres) and relative weight (in percent) were best for treatments three through five. In contrast, albumin height (in millimetres) and hu units improved significantly (p 0.05) for treatments four and five compared to the control therapy (treatment one).

Peculiarities of the Variation of Biologically Active Compounds in Fruit of Vaccinium oxycoccos L. Growing in the Čepkeliai State Strict Nature Reserve.

This study was carried out to analyze the accumulation patterns of anthocyanins, proanthocyanidins, flavonols, chlorogenic acid, and triterpene compounds in fruit samples of Vaccinium oxycoccos L. berries growing in the Čepkeliai State Strict Nature Reserve in Lithuania. Studies were carried out on the phytochemical composition of cranberry fruit samples during the period of 2020-2022. Anthocyanins, flavonols, chlorogenic acid and triterpene compounds were identified and quantified using UPLC-DAD methods, and proanthocyanins were determined using spectrophotometric methods. The content of identified compounds varied, as reflected in the total amounts of anthocyanins (710.3 ± 40 µg/g to 6993.8 ± 119 µg/g), proanthocyanidins (378.4 ± 10 µg EE/g to 3557. 3 ± 75 µg EE/g), flavonols (479.6 ± 9 µg/g to 7291.2 ± 226 µg/g), chlorogenic acid (68.0 ± 1 µg/g to 3858.2 ± 119 µg/g), and triterpenoids (3780.8 ± 98 µg/g to 7226.9 ± 224 µg/g). Cranberry fruit samples harvested from open oligotrophic wetland habitats contained higher levels of anthocyanins, anthocyanidins, flavonol glycosides, and proanthocyanidins. The highest levels of triterpene compounds were found in the cranberry fruits harvested in the spring of the following year after the snowmelt. The use of principal component analysis showed that cranberry plant material harvested in October and November had higher levels of bioactive compounds.

Transcriptome and weighted gene co-expression network analysis of jujube (Ziziphus jujuba Mill.) fruit reveal putative genes involved in proanthocyanin biosynthesis and regulation

Proanthocyanidin (PA) is an important bioactive compound with multiple physiological benefits in jujube (Ziziphus jujube Mill.). However, the molecular mechanisms underlying PA biosynthesis in jujube fruit have not been investigated. Here, the profiling of PA, (+)-catechin and (–)-epicatechin and transcriptome sequencing of three jujube cultivars from Xinjiang Uyghur Autonomous Region of China at five developmental stages were analyzed. The levels of total PAs and catechin exhibited a decreased trend over jujube ripening, and epicatechin content of two jujube cultivars increased first and then declined. Transcriptome analysis revealed that the differentially expressed genes (DEGs) were mainly enriched in ribosome, glycolysis/gluconeogenesis, fructose and mannose metabolism. 17 DEGs encoding PAL, CHS, CHI, CHS, F3’H, LAR, ANR, C4Hs, 4CLs, FLSs, DFRs and UFGTs involved in PA biosynthesis were relatively abundant. The highly transcribed LAR gene may greatly contribute to epicatechin accumulation. A weighted gene co-expression network analysis (WGCNA) was performed, and a network module including 1620 genes highly correlated with content of Pas and catechin was established. We identified 58 genes including 9 structural genes and 49 regulatory genes related to PA biosynthesis and regulation in the WGCNA module. Sixteen genes encoding 9 families of transcriptional factors (i.e., MYB, bHLH, ERF, bZIP, NAC, SBP, MIKC, HB, WRKY) were considered as hub genes. The results of qRT-PCR analysis validating 10 genes were well consistent with the transcriptome data. These findings provide valuable knowledge to facilitate its genetic studies and molecular breeding.

RhMYB1 and RhMYB123 form a positive feedback loop to regulate the proanthocyanidin biosynthesis in rose

Proanthocyanidins (PAs) are flavonoid-derived metabolites that make key contributions to plant and human health. Few studies have examined the mechanisms underlying the biosynthesis and accumulation of PAs in Rosa hybrida L. Here, we studied the role of the RhMYB1–RhMYB123 complex in regulating the biosynthesis of PAs in R. hybrida. The expression of PA biosynthesis genes was increased following overexpression of RhMYB123 in tobacco and rose somatic embryos, and this resulted in a marked increase in PA levels. Yeast two-hybrid assays showed that RhMYB123 interacted with RhMYB1, and these two proteins jointly promoted PAs biosynthesis by binding to the promoters of the genes encoding anthocyanin reductase (ANR) and leucoanthocyanidin reductase (LAR) for expression activation. Additional analysis showed that RhMYB1 could activate RhMYB123 expression through binding to related promoter and that the biosynthesis and accumulation of PAs are regulated by a positive feedback loop. Generally, the results of our study assist in better understanding the regulatory mechanisms of PAs biosynthesis from new perspectives and have implications for the use of metabolic engineering to enhance the quality of rose plants.

Phytochemical Composition of the Fruit of Large Cranberry (Vaccinium macrocarpon Aiton) Cultivars Grown in the Collection of the National Botanic Garden of Latvia

In this study, we conducted a qualitative and quantitative analysis of anthocyanins, proanthocyanidins, flavonols and triterpenoids in samples of introduced and bred large cranberry fruits from the collection of the National Botanic Garden of Latvia. The highest total anthocyanin levels (8638-9316 µg/g) were detected in the fruit samples of the cranberry cultivars 'Black Veil', 'Franclin' and 'Early Black'. The highest total proanthocyanidin levels (2775-3389 µg/g) were found in cranberries of cultivars 'Kalnciema Agrā', 'Kalnciema Tumšā', 'Searles', 'Howes', and 'Kalnciema Ražīgā'. The highest levels of flavonol compounds (1373-1402 µg/g) were detected in cranberries of cultivars 'Howes', 'Black Veil' and 'Salaspils Melnās'. The highest levels of triterpenoids (5292-5792 µg/g) were determined in cranberries of cultivars 'Kalnciema Agrā', 'Septembra', 'Džbrūklene' and 'Early Black'. The results of our study allow for the estimation of differences in the content of secondary metabolites in the fruit samples of the studied cranberry cultivars and for the selection of promising cultivars for further introduction and cultivation in the climatic conditions of the Baltic countries. These results are also important for the selection of the most promising cranberry cultivars for the preparation of cranberry raw material, and the high-quality composition of plant material ensures the effectiveness of cranberry supplements and other cranberry preparations.

Metabolite and Transcriptome Profiles of Proanthocyanidin Biosynthesis in the Development of Litchi Fruit.

The fruit of Litchi chinensis contains high levels of proanthocyanidins (PAs) in the pericarp. These substances can serve as substrates of laccase-mediated rapid pericarp browning after the fruit is harvested. In this study, we found that the major PAs in litchi pericarp were (-)-epicatechin (EC) and several procyanidins (PCs), primarily PC A2, B2, and B1, and the EC and the PC content decreased with the development of the fruit. RNA-seq analysis showed that 43 early and late structure genes related to flavonoid/PA biosynthesis were expressed in the pericarp, including five ANTHOCYANIDIN REDUCTASE (ANR), two LEUCOANTHOCYANIDIN REDUCTASE (LAR), and two ANTHOCYANIDIN SYNTHASE (ANS) genes functioning in the PA biosynthesis branch of the flavonoid pathway. Among these nine PA biosynthesis-related genes, ANR1a, LAR1/2, and ANS1 were highly positively correlated with changes in the EC/PC content, suggesting that they are the key PA biosynthesis-related genes. Several transcription factor (TF) genes, including MYB, bHLH, WRKY, and AP2 family members, were found to be highly correlated with ANR1a, LAR1/2, and ANS1, and their relevant binding elements were detected in the promoters of these target genes, strongly suggesting that these TF genes may play regulatory roles in PA biosynthesis. In summary, this study identified the candidate key structure and regulatory genes in PA biosynthesis in litchi pericarp, which will assist in understanding the accumulation of high levels of browning-related PA substances in the pericarp.

Bioactive compounds, anti-inflammatory, anti-nociceptive and antioxidant potentials of ethanolic leaf fraction of Sida linifolia L. (Malvaceae)

This study investigated the bioactive compounds, anti-inflammatory, anti-nociceptive, and antioxidant properties of the ethanolic leaf fraction of Sida linifolia (ELFSL). The in vitro anti-inflammatory study employed membrane stabilization, phospholipase A2, platelet aggregation, albumin denaturation, and protease inhibition assays. Intraperitoneal injection of freshly prepared carrageenan solution (0.1 mL of 0.01 g/mL), undiluted egg albumin (0.1 mL), acetic acid (0.6 % (v/v) (10 mL/kg bw), and formalin solution (0.02 mL of 1 % v/v) into mice hind paw, were used to evaluate the anti-inflammatory and anti-nociceptive mechanisms, respectively. In vitro antioxidant potentials were determined using 1,1-Diphenyl-2-picryl-hydrazyl (DPPH), nitric oxide (NO), ferric reducing power (FRAP), and total antioxidant capacity (TAC) assays. Varying quantities of flavonoids, phenols, tannins, saponins, terpenoids, steroids, and alkaloids, were detected in the fraction. GC-FID phytochemical profiling of ELFSL revealed a high level of epicatechin, moderate levels of catechin, kaempferol, flavone, naringenin, rutin flavanones, tannins, sapogenins, proanthocyanidin, and steroids, and small amounts of sparteine, resveratrol, and lunamarine. The ELFSL exerted excellent dose-dependent in vitro anti-inflammatory activities comparable with standard drugs (aspirin/prednisolone). The LD50 test showed safety up to 5000 mg/kg body weight (per oral) ELFSL. Interestingly, mice pre-administered various doses (200, 400, 600 mg/kg bw, po) of ELFSL showed significant (P < 0.05) reduction in edema, writhing, and time spent licking paw in all phases compared with control and were at par with 100 mg/kg bw (po) aspirin. The result also registered good concentration-dependent antioxidant potentials for ELFSL and was comparable to standards (gallic acid, butylated hydroxytoluene, and ascorbic acid). These imply that ELFSL possesses excellent antioxidant, anti-inflammatory, and anti-nociceptive potentials mediated by peripheral and central mechanisms.

Effects of in vitro digestion on protein degradation, phenolic compound release, and bioactivity of black bean tempeh.

The nutritional value and bioactivity of black beans are enhanced when fermented as tempeh, but their bioaccessibility and bioactivity after ingestion remain unclear. In this study, black bean tempeh and unfermented black beans were digested in vitro and changes in protein degradation, phenolic compound release, angiotensin I-converting enzyme (ACE)-inhibitory activity, and antioxidant activity between the two groups were compared. We observed that the soluble protein content of digested black bean tempeh was generally significantly higher than that of digested unfermented black beans at the same digestion stage (P < 0.05). The degree of protein hydrolysis and the content of <10 kDa peptides were also significantly higher in the digested black bean tempeh than in digested unfermented black beans (P < 0.05). SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and reversed-phase high-performance liquid chromatography (RP-HPLC) analysis showed that most macromolecular proteins in tempeh had been degraded during fermentation and more of the small peptides were released from black bean tempeh during digestion, respectively. Compared to that of the unfermented black beans, the level of ACE inhibition of black bean tempeh was lower, but this significantly increased to 82.51% following digestion, closing the gap with unfermented black beans. In addition, the total respective levels of phenolics, flavonoids, and proanthocyanidins released from black bean tempeh were 1.21, 1.40, and 1.55 times those of unfermented black beans following in vitro digestion, respectively. Antioxidant activity was also significantly higher in digested black bean tempeh than in digested unfermented black beans and showed a positive correlation with phenolic compound contents (P < 0.05). The results of this study proved that, compared to unfermented black beans, black bean tempeh retained protein and phenolic compound bioaccessibility and antioxidant activity and showed an improved ACE-inhibitory activity even after consumption.

Proanthocyanidins Alleviate Cadmium Stress in Industrial Hemp (Cannabis sativa L.).

Industrial hemp (Cannabis sativa L.), an annual herbaceous cash crop, is widely used for the remediation of heavy metal-contaminated soils due to its short growth cycle, high tolerance, high biomass, and lack of susceptibility to transfer heavy metals into the human food chain. In this study, a significant increase in proanthocyanidins was found in Yunnan hemp no. 1 after cadmium stress. Proanthocyanidins are presumed to be a key secondary metabolite for cadmium stress mitigation. Therefore, to investigate the effect of proanthocyanidins on industrial hemp under cadmium stress, four experimental treatments were set up: normal environment, cadmium stress, proanthocyanidin treatment, and cadmium stress after pretreatment with proanthocyanidins. The phenotypes from the different treatments were compared. The experimental results showed that pretreatment with proanthocyanidins significantly alleviated cadmium toxicity in industrial hemp. The transcriptome and metabolome of industrial hemp were evaluated in the different treatments. Proanthocyanidin treatment and cadmium stress in industrial hemp mainly affected gene expression in metabolic pathways associated with glutathione metabolism, phenylpropanoids, and photosynthesis, which in turn altered the metabolite content in metabolic pathways of phenylalanine, vitamin metabolism, and carotenoid synthesis. The combined transcriptomic and metabolomic analysis revealed that proanthocyanidins mitigated cadmium toxicity by enhancing photosynthesis, secondary metabolite synthesis, and antioxidant synthesis. In addition, exogenous proanthocyanidins and cadmium ions acted simultaneously on EDS1 to induce the production of large amounts of salicylic acid in the plant. Finally, overexpression of CsANR and CsLAR, key genes for proanthocyanidins synthesis in industrial hemp, was established in Arabidopsis plants. The corresponding plants were subjected to cadmium stress, and the results showed that CsLAR transgenic plants were more tolerant to cadmium than the CsANR transgenic and wild-type Arabidopsis plants. The results showed that salicylic acid and jasmonic acid were increased in Arabidopsis overexpressing CsLAR compared to AT wild-type Arabidopsis, and levels of secondary metabolites were significantly higher in Arabidopsis overexpressing CsLAR than in AT wild-type Arabidopsis. These results revealed how proanthocyanidins alleviated cadmium stress and laid the foundation for breeding industrial hemp varieties with higher levels of proanthocyanidins and greater tolerance.

The Impact of Storage Conditions and Bottle Orientation on the Evolution of Phenolic and Volatile Compounds of Vintage Port Wine.

This work evaluates the influence of the cellar conditions and bottle orientation, on the phenolic and volatile composition of a Vintage Port wine, sealed with natural cork stoppers, for 44 months post-bottling. The storage was performed in two different cellars, namely a cellar A with controlled temperature and humidity, and a cellar B, representing a traditional cellar, with uncontrolled temperature and humidity. The impact of bottle orientation was studied in cellar A, where the bottles were stored in horizontal and vertical positions. The phenolic and volatile composition of the bottled Vintage Port wine were analyzed after 6, 15 and 44 months. The results unveiled that the cellar conditions and bottle orientation had an impact in Port wine composition which was higher at 44 months post-bottling. The samples stored in the traditional cellar unveiled significantly higher yellow tones, lower tannin specific activity, and higher levels of furfural and 5-methylfurfural. Furthermore, the samples stored in the horizontal position revealed significant higher levels of total proanthocyanidins and higher tannin specific activity than the samples stored in the vertical position. Interestingly, for the first time to our knowledge, an ellagitannin-derived compound (Corklin) was detected in Vintage Port wines stored in the horizontal position, which results from the reaction of cork constituents with phenolic compounds present in wines.

Prophylactic Effects of Purple Shoot Green Tea on Cytokine Immunomodulation through Scavenging Free Radicals and NO in LPS-Stimulated Macrophages

Polyphenols and flavonoids from non-fermented green tea and fully-fermented black tea exhibit antioxidant abilities that function as natural health foods for daily consumption. Nonetheless, evidence regarding prophylactic effects of purple shoot tea on immunomodulation remains scarce. We compared the immunomodulatory effects of different tea processes on oxidative stress and cytokine expressions in lipopolysaccharide (LPS)-stimulated macrophages. Major constituents of four tea products, Taiwan Tea Experiment Station No.12 (TTES No. 12) black and green tea and purple shoot black and purple shoot green tea (TB, TG, PB and PG, respectively), were analyzed to explore the prophylactic effects on expressions of free radicals, nitric oxide (NO), monocyte chemoattractant protein-1 (MCP-1), interleukin 6 (IL-6) and tumor necrosis factor α (TNF-α) in LPS-activated RAW264.7 cell models. PG contained abundant levels of total polyphenols, flavonoids, condensed tannins and proanthocyanidins (371.28 ± 3.83; 86.37 ± 1.46; 234.67 ± 10.1; and 24.81 ± 0.75 mg/g, respectively) contributing to excellent free radical scavenging potency. In both the LPS-activated inflammation model and the prophylactic model, all tea extracts suppressed NO secretion in a dose-dependent manner, especially for PG. Intriguingly, most tea extracts enhanced expressions of IL-6 in LPS-stimulated macrophages, except PG. However, all teas disrupted downstream transduction of chemoattractant MCP-1 for immune cell trafficking. In the prophylactic model, all teas inhibited inflammatory responses by attenuating expressions of IL-6 and TNF-α in a dose-dependent manner, especially for TG and PG. Our prophylactic model demonstrated PG exerts robust effects on modulating LPS-induced cytokine expressions of MCP-1, IL-6 and TNF-α through scavenging free radicals and NO. In light of the prophylactic effects on LPS-related inflammation, PG effectively scavenges free radicals to modulate cytokine cascades that could serve as a functional beverage for immunomodulation.

BnbHLH92a negatively regulates anthocyanin and proanthocyanidin biosynthesis in Brassica napus

Yellow seed trait is a desirable characteristic with potential for increasing seed quality and commercial value in rapeseed, and anthocyanin and proanthocyanidins (PAs) are major seed-coat pigments. Few transcription factors involved in the regulation of anthocyanin and PAs biosynthesis have been characterized in rapeseed. In this study, we identified a transcription factor gene BnbHLH92a (BnaA06T0441000ZS) in rapeseed. Overexpressing BnbHLH92a both in Arabidopsis and in rapeseed reduced levels of anthocyanin and PAs. Correspondingly, the expression profiles of anthocyanin and PA biosynthesis genes (TT3, BAN, TT8, TT18, and TTG1) were shown by quantitative real-time PCR to be inhibited in BnbHLH92a-overexpressing Arabidopsis seeds, indicating that BnbHLH92a represses the anthocyanin and PA biosynthesis pathway in Arabidopsis. BnbHLH92a physically interacts with the BnTTG1 protein and represses the biosynthesis of anthocyanins and PAs in rapeseed. BnbHLH92a also binds directly to the BnTT18 promoter and represses its expression. These results suggest that BnbHLH92a is a novel upstream regulator of flavonoid biosynthesis in B. napus.