Transcriptional and physiological analyses uncover the mineralization and uptake mechanisms of phytic acid in symbiotically grown Vicia faba plants.

Legume-rhizobia symbiosis requires high phosphorus (P) in the form of ATP to convert atmospheric nitrogen (N) into ammonia. The fixed ammonia is converted to NH4+ by H+-ATPase via protonation. To the best of our knowledge, most of these research works resort to using only inorganic P (Pi) to the neglect of the organic P (Po) counterpart. As it stands, the potential regulating roles of plasma membrane (PM) H+-ATPases during legume-rhizobia symbiosis in response to phytic acid supply and how it alters and modulates the regulation of PM H+-ATPases remain obscure. To contribute to the above hypothesis, we investigate the mechanisms that coordinately facilitate the growth, uptake, and transcript expression of PM H+-ATPase gene isoforms in response to different P sources when hydroponically grown Vicia faba plants were exposed to three P treatments, viz., low- and high-Pi (2.0 and 200 µM KH2PO4; LPi and HPi), and phytic acid (200 µM; Po) and inoculated with Rhizobium leguminosarum bv. viciae 384 for 30 days. The results consistently reveal that the supply of Po improved not only the growth and biomass, but also enhanced photosynthetic parameters, P uptake and phosphatase activities in symbiotically grown Vicia faba relative to Pi. The supply of Po induced higher transcriptional expression of all PM H+-ATPase gene isoforms, with possible interactions between phosphatases and H+-ATPase genes in Vicia faba plants when exclusively reliant on N derived from nodule symbiosis. Overall, preliminary results suggest that Po could be used as an alternative nutrition in symbiotic crops to improve plant growth.

Exploring the biochemical dynamics in faba bean (Vicia faba L. minor) in response to Orobanche foetida Poir. parasitism under inoculation with different rhizobia strains.

In Tunisia, Orobanche foetida Poir. is considered an important agricultural biotic constraint on faba bean (Vicia faba L.) production. An innovative control method for managing this weed in faba bean is induced resistance through inoculation by rhizobia strains. In this study, we explored the biochemical dynamics in V. faba L. minor inoculated by rhizobia in response to O. foetida parasitism. A systemic induced resistant reaction was evaluated through an assay of peroxidase (POX), polyphenol oxidase (PPO) and phenyl alanine ammonialyase (PAL) activity and phenolic compound and hydrogen peroxide (H2O2) accumulation in faba bean plants infested with O. foetida and inoculated with rhizobia. Two rhizobia strains (Mat, Bj1) and a susceptible variety of cultivar Badi were used in a co-culture Petri dish experiment. We found that Mat inoculation significantly decreased O. foetida germination and the number of tubercles on the faba bean roots by 87% and 88%, respectively. Following Bj1 inoculation, significant decreases were only observed in O. foetida germination (62%). In addition, Mat and Bj1 inoculation induced a delay in tubercle formation (two weeks) and necrosis in the attached tubercles (12.50% and 4.16%, respectively) compared to the infested control. The resistance of V. faba to O. foetida following Mat strain inoculation was mainly associated with a relatively more efficient enzymatic antioxidative response. The antioxidant enzyme activity was enhanced following Mat inoculation of the infected faba bean plant. Indeed, increases of 45%, 67% and 86% were recorded in the POX, PPO and PAL activity, respectively. Improvements of 56% and 12% were also observed in the soluble phenolic and H2O2 contents. Regarding inoculation with the Bj1 strain, significant increases were only observed in soluble phenolic and H2O2 contents and PPO activity (especially at 45 days after inoculation) compared to the infested control. These results imply that inoculation with the rhizobia strains (especially Mat) induced resistance and could bio-protect V. faba against O. foetida parasitism by inducing systemic resistance, although complete protectionwas not achieved by rhizobia inoculation. The Mat strain could be used as a potential candidate for the development of an integrated method for controlling O. foetida parasitism in faba bean.

Green-synthesized zinc oxide nanoparticles by Enterobacter sp.: unveiling characterization, antimicrobial potency, and alleviation of copper stress in Vicia faba (L.) plants.

BACKGROUND: The biosynthesis of zinc oxide nanoparticles (ZnO NPs) using Enterobacter sp. and the evaluation of their antimicrobial and copper stress (Cu+ 2)-reducing capabilities in Vicia faba (L.) plants. The green-synthesized ZnO NPs were validated using X-ray powder diffraction (XRD); Fourier transformed infrared (FTIR), Ultraviolet-Visible spectroscopy (UV-Vis), Transmission electron microscope (TEM) and scanning electron microscopy (SEM) techniques. ZnO NPs could serve as an improved bactericidal agent for various biological applications. as well as these nanoparticles used in alleviating the hazardous effects of copper stress on the morphological and physiological traits of 21-day-old Vicia faba (L.) plants. RESULTS: The results revealed that different concentrations of ZnO NPs (250, 500, or 1000 mg L-1) significantly alleviated the toxic effects of copper stress (100 mM CuSO4) and increased the growth parameters, photosynthetic efficiency (Fv/Fm), and pigments (Chlorophyll a and b) contents in Cu-stressed Vicia faba (L.) seedlings. Furthermore, applying high concentration of ZnO NPs (1000 mg L-1) was the best dose in maintaining the levels of antioxidant enzymes (CAT, SOD, and POX), total soluble carbohydrates, total soluble proteins, phenolic and flavonoid in all Cu-stressed Vicia faba (L.) seedlings. Additionally, contents of Malondialdehyde (MDA) and hydrogen peroxide (H2O2) were significantly suppressed in response to high concentrations of ZnO NPs (1000 mg L-1) in all Cu-stressed Vicia faba (L.) seedlings. Also, it demonstrates strong antibacterial action (0.9 mg/ml) against various pathogenic microorganisms. CONCLUSIONS: The ZnO NPs produced in this study demonstrated the potential to enhance plant detoxification and tolerance mechanisms, enabling plants to better cope with environmental stress. Furthermore, these nanoparticles could serve as an improved bactericidal agent for various biological applications.

Low-intensity pulsed electric field processing prior to germination improves in vitro digestibility of faba bean (Vicia faba L.) flour and its derived products: A case study on legume-enriched wheat bread.

This study investigated the effect of low-intensity pulsed electric field (PEF) (0.3-0.7 kV/cm) and/or germination (0-72 h, 20 °C) on faba beans prior to flour- and breadmaking. PEF (0.5 and 0.7 kV/cm) had no significant effect on the germination performance of faba bean but had a positive effect on in vitro starch and protein hydrolysis of PEF-treated beans germinated for 72 h. The incorporation of flour from soaked, germinated, PEF-treated, and PEF-treated+germinated faba beans into wheat bread, at 30% mass level, improved the nutritional composition (total starch and protein contents) and protein digestibility but it reduced the specific volume and increased the density, brownness, and hardness of the bread. This finding shows for the first time that PEF-treatment (<0.7 kV/cm) of faba beans followed by germination (72 h) improved in vitro starch and protein hydrolysis of its flour and the protein digestibility at gastric phase of its enriched wheat bread.

Antioxidant and Angiotensin-Converting Enzyme Inhibitory Activity of Faba Bean-Derived Peptides After In Vitro Gastrointestinal Digestion: Insight into Their Mechanism of Action.

Faba bean flour, after in vitro gastrointestinal digestion, showed important antioxidant and angiotensin-converting enzyme (ACE) inhibitory activities. In the present study, 11 faba bean- derived peptides were synthesized to confirm their bioactivities and provide a deeper understanding of their mechanisms of action. The results revealed that 7 peptides were potent antioxidants, namely, NYDEGSEPR, TETWNPNHPEL, TETWNPNHPE, VIPTEPPH, VIPTEPPHA, VVIPTEPPHA, and VVIPTEPPH. Among them, TETWNPNHPEL had the highest activity in the ABTS (EC50 = 0.5 ± 0.2 mM) and DPPH (EC50 = 2.1 ± 0.1 mM) assays (p < 0.05), whereas TETWNPNHPE had the highest activity (p < 0.05) in the ORAC assay (2.84 ± 0.08 mM Trolox equivalent/mM). Synergistic and/or additive effects were found when selected peptides (TETWNPNHPEL, NYDEGSEPR, and VVIPTEPPHA) were combined. Four peptides were potent ACE inhibitors, where VVIPTEPPH (IC50 = 43 ± 1 µM) and VVIPTEPPHA (IC50 = 50 ± 5 µM) had the highest activity (p < 0.05), followed by VIPTEPPH (IC50 = 90 ± 10 µM) and then VIPTEPPHA (IC50 = 123 ± 5 µM) (p < 0.05). These peptides were noncompetitive inhibitors, as supported by kinetic studies and a molecular docking investigation. This study demonstrated that peptides derived from faba beans have multifunctional bioactivities, making them a promising food-functional and nutraceutical ingredient.

Preparation of isolated guard cells, containing cell walls, from Vicia faba.

Stomatal movement, initiated by specialized epidermal cells known as guard cells (GCs), plays a pivotal role in plant gas exchange and water use efficiency. Despite protocols existing for isolating GCs through proplasting for carrying out biochemical, physiological, and molecular studies, protocals for isolating GCs with their cell walls still intact have been lacking in the literature. In this paper, we introduce a method for the isolation of complete GCs from Vicia faba and show their membrane to remain impermeable through propidium iodide staining. This methodology enables further in-depth analyses into the cell wall composition of GCs, facilitating our understanding of structure-function relationship governing reversible actuation within cells.

True Ileal Amino Acid Digestibility and Protein Quality of 15N-Labeled Faba Bean in Healthy Humans.

BACKGROUND: The recommended transition toward more plant-based diets, particularly containing legumes, requires a wider knowledge of plant protein bioavailability. Faba beans are cultivated at different latitudes and are used increasingly in human nutrition. OBJECTIVES: We aimed to assess the nutritional quality of faba bean protein in healthy volunteers equipped with an intestinal tube to implement the ileal 15N balance method. METHODS: Nine volunteers completed the study (7 males, 2 females, aged 33 ± 10 y, BMI: 24.7 ± 2.6 kg/m2). They were equipped with a nasoileal tube. After fasting overnight, they ingested a test meal consisting of cooked mash of dehulled faba bean seeds (20 g protein per serving of approximately 250 g) intrinsically labeled with 15N. Samples of ileal contents, plasma, and urine were collected over an 8-h postprandial period. Undigested nitrogen (N) and amino acids (AAs) were determined using isotopic MS, and subsequently, ileal digestibility and digestible indispensable amino acid score (DIAAS) were calculated. The measurement of postprandial deamination allowed calculation of the net postprandial protein utilization (NPPU). RESULTS: The ileal N digestibility was 84.1% ± 7.7%. Postprandial deamination represented 19.2% ± 3.6% of ingested N, and the NPPU was 64.7% ± 9.7%. The ileal digestibility of individual AAs varied from 85.1% ± 13.7% for histidine to 94.2% ± 3.6% for glutamine + glutamate. The mean AA digestibility was ∼6 percentage points higher than the digestibility of N, reaching 89.8% ± 5.9%, whereas indispensable AA digestibility was 88.0% ± 7.3%. Histidine and tryptophan were the first limiting AAs [DIAAS = 0.77 (calculated by legume-specific N-to-protein conversion factor 5.4); 0.67 (by default factor 6.25)]. Sulfur AAs were limiting to a lesser extent [DIAA ratio = 0.94 (N × 5.4); 0.81 (N × 6.25)]. CONCLUSIONS: Protein ileal digestibility of cooked, dehulled faba beans in humans was moderate (<85%), but that of AAs was close to 90%. Overall protein quality was restricted by the limited histidine and tryptophan content. This trial was registered at clinicaltrials.gov as NCT05047757.

Amelioration of NaCl stress on germination, growth, and nitrogen fixation of Vicia faba at isosmotic Na-Ca combinations and Rhizobium.

MAIN CONCLUSION: The Na+/Ca2+ ratio of 1/5 ameliorated the inhibitory action of NaCl and improved the germination and growth of Vicia faba. Addition of Rhizobium also enhanced nodulation and nitrogen fixation. Casting light upon the impact of salinity stress on growth and nitrogen fixation of Vicia faba supplemented with Rhizobium has been traced in this work. How Ca2+ antagonizes Na+ toxicity and osmotic stress of NaCl was also targeted in isosmotic combinations of NaCl and CaCl2 having various Na+:Ca2+ ratios. Growth of Vicia faba (cultivar Giza 3) was studied at two stages: germination and seedling. At both experiments, seeds or seedlings were exposed to successively increasing salinity levels (0, 50, 100, 150, and 200 mM NaCl) as well as isosmotic combinations of NaCl and CaCl2 (Na+:Ca2+ of 1:1, 1:5, 1:10, 1:15, 1:18, and 1: 20), equivalent to 150 mM NaCl. Inocula of the local nitrogen-fixing bacteria, Rhizobium leguminosarum (OP715892) were supplemented at both stages. NaCl salinity exerted a negative impact on growth and metabolism of Vicia faba; inhibition was proportional with increasing salinity level up to the highest level of 200 mM. Seed germination, shoot and root lengths, fresh and dry weights, chlorophyll content, and nodules (number, weight, leghemoglobin, respiration, and nitrogenase activity) were inhibited by salinity. Ca2+ substitution for Na+, particularly at a Na/Ca ratio of 1:5, was stimulatory to almost all parameters at both stages. Statistical correlations between salinity levels and Na/Ca combinations proved one of the four levels (strong- or weak positive, strong- or weak negative) with most of the investigated parameters, depending on the parameter.

Thaumatin-like protein family genes VfTLP4-3 and VfTLP5 are critical for faba bean's response to drought stress at the seedling stage.

Thaumatin-like proteins (TLPs) are a diverse family of pathogenesis-related proteins (PR-5) found in various plant species. Faba bean is an economically important crop known for its nutritional value and resilience to harsh environmental conditions, including drought. In this study, we conducted a comprehensive analysis of the gene structure, phylogenetics, and expression patterns of TLP genes in faba bean, with a specific focus on their response to drought stress. A total of 10 TLP genes were identified and characterized from the faba bean transcriptome, which could be classified into four distinct groups based on their evolutionary relationships. Conserved cysteine residues and REDDD motifs, which are characteristic features of TLPs, were found in most of the identified VfTLP members, and these proteins were likely to reside in the cytoplasm. Two genes, VfTLP4-3 and VfTLP5, exhibited significant upregulation under drought conditions. Additionally, ectopically expressing VfTLP4-3 and VfTLP5 in tobacco leaves resulted in enhanced drought tolerance and increased peroxidase (POD) activity. Moreover, the protein VfTLP4-3 was hypothesized to interact with glycoside hydrolase family 18 (GH18), endochitinase, dehydrin, Barwin, and aldolase, all of which are implicated in chitin metabolism. Conversely, VfTLP5 was anticipated to associate with peptidyl-prolyl cis-trans isomerase-like 3, a molecule linked to the synthesis of proline. These findings suggest that these genes may play crucial roles in mediating the drought response in faba bean through the regulation of these metabolic pathways, and serve as a foundation for future genetic improvement strategies targeting enhanced drought resilience in this economically important crop.

From ancient crop to modern superfood: Exploring the history, diversity, characteristics, technological applications, and culinary uses of Peruvian fava beans.

The search for plant-based superfoods has shown that many regional populations already have these foods in their diet, with significant potential for production and marketing. This critical review intends to show the history, diversity, characteristics, and uses, emphasizing their significance in traditional diets and potential in the food industry of Peruvian fava beans. As a valuable plant-based protein source, fava beans offer essential micronutrients and have diverse culinary applications. Innovative food industry applications include plant-based meat alternatives, fortified gluten-free products, and a natural color, protein, and fiber source in extruded foods. Key studies have highlighted the successful incorporation of fava beans into various food products, improving their nutritional properties, though some studies also point to limitations in their sensory acceptance. Further research is needed to understand the bioactive components, health effects, and techno-functional characteristics of beans. Challenges facing cultivating and consuming fava beans in Peru include adapting to climate change, enhancing productivity and quality, and promoting consumption and added value. Addressing these challenges involves developing climate-resilient varieties, optimizing agricultural practices, and providing access to resources and financing. In conclusion, this review highlights the promising prospects of Peruvian fava beans as a sustainable, nutritionally rich, and versatile ingredient in the food industry. By harnessing their potential and overcoming challenges, Peruvian fava beans can transition from an ancient crop to a modern superfood, inspiring a global shift towards sustainable and nutritionally balanced diets, aiding the fight against malnutrition, and enriching culinary traditions worldwide.

An Investigation of the Protein Quality and Temporal Pattern of Peripheral Blood Aminoacidemia following Ingestion of 0.33 g·kg-1 Body Mass Protein Isolates of Whey, Pea, and Fava Bean in Healthy, Young Adult Men.

An increase in the intake of legumes is recommended in the promotion of plant-sourced (PSP) rather than animal-sourced (ASP) protein intake to produce a more sustainable diet. This study evaluated the quality of novel PSP isolates from pea (PEA) and fava bean (FAVA) and an ASP isolate of whey (WHEY) and compared the magnitude and temporal pattern of peripheral arterial aminoacidemia following ingestion of 0.33 g·kg-1 body mass of protein isolate in healthy young adult men (n = 9). Total indispensable amino acids (IAA) comprised 58% (WHEY), 46% (PEA), and 42% (FAVA) of the total amino acid (AA) composition, with the ingested protein providing 108% (WHEY), 77% (PEA), and 67% (FAVA) of the recommended per diem requirement of IAA. Reflecting the AA composition, the area under the curve (∆AUC0-180), post-ingestion increase in total IAA for WHEY was 41% (p < 0.001) and 57% (p < 0.001) greater than PEA and FAVA, respectively, with PEA exceeding FAVA by 28% (p = 0.003). As a sole-source, single-dose meal-size serving, the lower total IAA for PEA and FAVA would likely evoke a reduced post-prandial anabolic capacity compared to WHEY. Incorporated into a food matrix, the promotion of PSP isolates contributes to a more sustainable diet.

Epigenetic modifications evidenced by isolation of proteins on nascent DNA and immunofluorescence in hydroxyurea-treated root meristem cells of Vicia faba.

MAIN CONCLUSION: By implementation of the iPOND technique for plant material, changes in posttranslational modifications of histones were identified in hydroxyurea-treated root meristem cells of Vicia. Replication stress (RS) disrupts or inhibits replication forks and by altering epigenetic information of the newly formed chromatin can affect gene regulation and/or spatial organisation of DNA. Experiments on Vicia faba root meristem cells exposed to short-term treatment with 3 mM hydroxyurea (HU, an inhibitor of DNA replication) were aimed to understand epigenetic changes related to RS. To achieve this, the following histone modifications were studied using isolation of proteins on nascent DNA (iPOND) technique (for the first time on plant material) combined with immunofluorescence labeling: (i) acetylation of histone H3 at lysine 56 (H3K56Ac), (ii) acetylation of histone H4 at Lys 5 (H4K5Ac), and (iii) phosphorylation of histone H3 at threonine 45 (H3T45Ph). Certainly, the implementation of the iPOND method for plants may prove to be a key step for a more in-depth understanding of the cell's response to RS at the chromatin level.

Protein blends and extrusion processing to improve the nutritional quality of plant proteins.

Plant proteins have low protein nutritional quality due to their unbalanced indispensable amino acid (IAA) profile and the presence of antinutritional factors (ANFs) that limit protein digestibility. The blending of pulses with cereals/pseudocereals can ensure a complete protein source of IAA. In addition, extrusion may be an effective way to reduce ANFs and improve protein digestibility. Thereby, we aimed to improve the protein nutritional quality of plant protein ingredients by blending different protein sources and applying extrusion processing. Protein blends were prepared with pea, faba bean, quinoa, hemp, and/or oat concentrates or flours, and extrudates were prepared either rich in pulses (texturized vegetable proteins, TVPs) or rich in cereals (referred to here as Snacks). After extrusion, all samples showed a reduction in trypsin inhibitor activity (TIA) greater than 71%. Extrusion caused an increase in the total in vitro protein digestibility (IVPD) of TVPs, whereas no significant effect was shown for the snacks. According to the molecular weight distribution, TVPs presented protein aggregation. The results suggest that the positive effect of decreased TIA on IVPD is partially counteracted by the formation of aggregates during extrusion which restricts enzyme accessibility. After extrusion, all snacks retained a balanced amino acid score whereas a small loss of methionine + cysteine was observed in the TVPs, resulting in a small reduction in IAA content. Thus, extrusion has the potential to improve the nutritional quality of TVPs by reducing TIA and increasing protein digestibility.

Hormonal crosstalk controls cell death induced by kinetin in roots of Vicia faba ssp. minor seedlings.

Studies of vitality/mortality of cortex cells, as well as of the concentrations of ethylene (ETH), gibberellins (GAs), indolic compounds/auxins (ICs/AUXs) and cytokinins (CKs), were undertaken to explain the hormonal background of kinetin (Kin)-regulated cell death (RCD), which is induced in the cortex of the apical parts of roots of faba bean (Vicia faba ssp. minor) seedlings. Quantification was carried out with fluorescence microscopy, ETH sensors, spectrophotometry and ultrahigh-performance liquid chromatography tandem mass spectrometry (UHPLC‒MS/MS). The results indicated that Kin was metabolized to the transport form, i.e., kinetin-9-glucoside (Kin9G) and kinetin riboside (KinR). KinR was then converted to cis-zeatin (cZ) in apical parts of roots with meristems, to cis-zeatin riboside (cZR) in apical parts of roots without meristems and finally to cis-zeatin riboside 5'-monophosphate (cZR5'MP), which is indicated to be a ligand of cytokinin-dependent receptors inducing CD. The process may be enhanced by an increase in the amount of dihydrozeatin riboside (DHZR) as a byproduct of the pathway of zeatin metabolism. It seems that crosstalk of ETH, ICs/AUXs, GAs and CKs with the cZR5'MP, the cis-zeatin-dependent pathway, but not the trans-zeatin-dependent pathway, is responsible for Kin-RCD, indicating that the process is very specific and offers a useful model for studies of CD hallmarks in plants.

Vicia faba Peptide Network Supplementation Does Not Differ From Milk Protein in Modulating Changes in Muscle Size During Short-Term Immobilization and Subsequent Remobilization, but Increases Muscle Protein Synthesis Rates During Remobilization in Healthy Young Men.

BACKGROUND: Muscle mass and strength decrease during short periods of immobilization and slowly recover during remobilization. Recent artificial intelligence applications have identified peptides that appear to possess anabolic properties in in vitro assays and murine models. OBJECTIVES: This study aimed to compare the impact of Vicia faba peptide network compared with milk protein supplementation on muscle mass and strength loss during limb immobilization and regain during remobilization. METHODS: Thirty young (24 ± 5 y) men were subjected to 7 d of one-legged knee immobilization followed by 14 d of ambulant recovery. Participants were randomly allocated to ingest either 10 g of the Vicia faba peptide network (NPN_1; n = 15) or an isonitrogenous control (milk protein concentrate; MPC; n = 15) twice daily throughout the study. Single-slice computed tomography scans were performed to assess quadriceps cross-sectional area (CSA). Deuterium oxide ingestion and muscle biopsy sampling were applied to measure myofibrillar protein synthesis rates. RESULTS: Leg immobilization decreased quadriceps CSA (primary outcome) from 81.9 ± 10.6 to 76.5 ± 9.2 cm2 and from 74.8 ± 10.6 to 71.5 ± 9.8 cm2 in the NPN_1 and MPC groups, respectively (P < 0.001). Remobilization partially recovered quadriceps CSA (77.3 ± 9.3 and 72.6 ± 10.0 cm2, respectively; P = 0.009), with no differences between the groups (P > 0.05). During immobilization, myofibrillar protein synthesis rates (secondary outcome) were lower in the immobilized leg (1.07% ± 0.24% and 1.10% ± 0.24%/d, respectively) than in the non-immobilized leg (1.55% ± 0.27% and 1.52% ± 0.20%/d, respectively; P < 0.001), with no differences between the groups (P > 0.05). During remobilization, myofibrillar protein synthesis rates in the immobilized leg were greater with NPN_1 than those with MPC (1.53% ± 0.38% vs. 1.23% ± 0.36%/d, respectively; P = 0.027). CONCLUSION: NPN_1 supplementation does not differ from milk protein in modulating the loss of muscle size during short-term immobilization and the regain during remobilization in young men. NPN_1 supplementation does not differ from milk protein supplementation in modulating the myofibrillar protein synthesis rates during immobilization but further increases myofibrillar protein synthesis rates during remobilization.

Electron beam irradiation modification of ultra-high pressure treated broad bean starch: Improvement of multi-scale structure and functional properties.

To investigate the synergistic effect of electron beam irradiation (EBI) on the ultra-high pressure (UHP) modification of broad bean starch, various pressures (200, 400, 600 MPa) combined with different irradiation doses (3, 6, 12 kGy) were used to modify the structure-properties of broad bean starch in this study. The results showed that both UHP and EBI induced a reduction of amylopectin molecular weight (Mw) and depolymerization of long chains, caused the loss of short-range ordered structure and imperfection of crystal structure, and improved starch viscosity, solubility and enzyme sensitivity. Furthermore, the applied pressure causes changes in starch granule structure, upon which EBI promotes further degradation and depolymerization of starch by affecting the crystalline and amorphous regions. Hence, appropriate doses of EBI treatment can impart more desirable processing properties to UHP-modified starches, and EBI can be used as a promising way to promote starch modification further.

A comparative metabolomics investigation of flavonoid variation in faba bean flowers.

INTRODUCTION: Faba bean (Vicia faba L.) flowers are edible and used as garnishes because of their aroma, sweet flavor and attractive colors. Anthocyanins are the common plant pigments that give flowers their vivid colors, whereas non-anthocyanin flavonoids can serve as co-pigments that can modify the color intensity of flowers. OBJECTIVES: To explore the polyphenol diversity and differences in standard and wing petals of faba bean flowers; and identify glycosylated flavonoids that contribute to flower color. METHODS: Flower standard and wing petals from 30 faba bean genotypes (eight color groups with a total of 60 samples) were used for polyphenol extraction. Samples were analyzed using a targeted method and a semi-untargeted analysis using liquid chromatography-high resolution mass spectrometry (LC-HRMS) combined with photodiode array (PDA) detection. Compound Discoverer software was used for polyphenol identification and multivariate analysis. RESULTS: The semi-untargeted analysis guided by the PDA detected 90 flavonoid metabolites present in faba bean flower petals. Ten anthocyanins largely influenced the flower colors, but other flavonoids (63 flavonols and 12 flavones) found with variable levels in different flower color groups appeared to also influence color, especially in mixed colors. CONCLUSION: Analysis of the different colored faba bean flowers confirmed that the color variation between the flowers was mainly controlled by anthocyanins in brown, red and purple-red flowers. Of the other flavonoids, multiglycosylated kaempferols were abundant in white and brown flowers, monoglycosylated kaempferols were common in red and purple-red flowers, and quercetin and apigenin glycosides were abundant co-pigments in purple-red flowers.

Effects of forage and grain legume-based silages supplemented with faba bean meal or rapeseed expeller on lactational performance, nitrogen utilization, and plasma amino acids in dairy cows.

The objective of this experiment was to investigate the effect of forage type [red clover (51%)-grass silage, i.e., RCG; vs. faba bean (66%)-grass silage, i.e., FBG] and concentrate type (faba bean, FB; vs. rapeseed expeller, RE) on lactational performance, milk composition and nitrogen (N) utilization in lactating dairy cows. Eight lactating multiparous Nordic Red cows were used in a replicated 4 × 4 Latin Square experiment, with 21-d periods, in a 2 × 2 factorial arrangement of treatments. The experimental treatments were as follows: (1) RCG with RE, (2) RCG with FB, (3) FBG with RE, and (4) FBG with FB. Inclusion rates of RE and FB were isonitrogenous. Crude protein contents of the experimental diets were 16.3, 15.9, 18.1, and 17.9% of dry matter, respectively. All diets included oats and barley and were fed ad libitum as total mixed rations with forage-to-concentrate ratio of 55:45. Dry matter intake and milk yield were recorded daily, and spot samples of urine, feces, and blood were collected at the end of each experimental period. Dry matter intake did not differ across diets, averaging 26.7 kg/d. Milk yield averaged 35.6 kg/d and was 1.1 kg/d greater for RCG versus FBG, and milk urea N concentration was lower for RCG compared with FBG. Milk yield was 2.2 kg/d and milk protein yield 66 g/d lower for FB versus RE. Nitrogen intake, urinary N, and urinary urea N excretions were lower, and milk N excretion tended to be lower for RCG compared with FBG. The proportion of the dietary N excreted as fecal N was larger in cows fed RCG than for those fed FBG, and the opposite was true for urinary N. We detected an interaction for milk N as percentage of N intake: it increased with RE compared with FB for RCG-based diet, but only a marginal increase was observed for FBG-based diet. Plasma concentration of His and Lys were lower for RCG than for FBG, whereas His tended to be greater and Lys lower for FB compared with RE. Further, plasma Met concentration was around 26% lower for FB than for RE. Of milk fatty acids, saturated fatty acids were decreased by RCG and increased by FB compared with FBG and RE, respectively, whereas monounsaturated fatty acids were increased by RCG versus FBG, and were lower for FB than for RE. In particular, 18:1n-9 concentration was lower for FB compared with RE. Polyunsaturated fatty acids, such as 18:2n-6 and 18:3n-3, were greater for RCG than for FBG, and 18:2n-6 was greater and 18:3n-3 was lower for FB versus RE. In addition, cis-9,trans-11 conjugated linoleic acid was lower for FB compared with RE. Faba bean whole-crop silage and faba bean meal have potential to be used as a part of dairy cow rations, but further research is needed to improve their N efficiency. Red clover-grass silage from a mixed sward, without inorganic N fertilizer input, combined with RE, resulted in the greatest N efficiency in the conditions of this experiment.

Comparison of standardized ileal digestibility of amino acids in faba beans and field peas in broiler chickens and pigs.

An experiment, using the same set of five diets was conducted to determine and compare the standardized ileal digestibility (SID) of amino acids (AA) in faba beans and three cultivars of field peas in broiler chickens and pigs. Four test diets were prepared to contain faba beans, DS-Admiral field peas, Hampton field peas, or 4010 field peas as the sole source of nitrogen. A nitrogen-free diet (NFD), as the fifth diet, was prepared to estimate the basal endogenous losses of AA to determine the SID of AA in the test ingredients. A total of 416 male broiler chickens with an initial body weight (BW) of 951 ±â€…111 g were allotted to five diets in a randomized complete block design with BW as a blocking factor on day 21 posthatching. There were 8 replicate cages with 10 birds per cage for diets containing test ingredients and 12 birds per cage for NFD. All birds were allowed ad-libitum access to feed for 5 days. On day 26 posthatching, all birds were euthanized by CO2 asphyxiation and digesta contents were collected from the distal two-thirds of the ileum. Twenty barrows with an initial BW of 30.2 ±â€…1.58 kg, surgically fitted with T-cannulas in the distal ileum, were divided into four blocks according to BW and each block was assigned to a 5 × 2 incomplete Latin Square design that consisted of five dietary treatments and two experimental periods. For each experimental period, there was a 5-day adaptation period followed by a 2-day collection of ileal digesta samples. The data were analyzed as a 2 × 4 factorial treatment arrangement with the effect of species (i.e., broiler chickens and pigs) and test diets (i.e., four test ingredients). For broiler chickens, the SID of Lys in faba beans, DS-Admiral field peas, and Hampton field peas were above 90% but in 4010 field peas, it was 85.1%. The SID of Lys in faba beans, DS-Admiral field peas, and Hampton field peas for pigs was above 80% but was 78.9% in 4010 field peas. The respective SID of Met in faba beans, DS-Admiral field peas, Hampton field peas, and 4010 field peas were 84.1%, 87.3%, 89.8%, and 72.1% for broiler chickens, and 71.5%, 80.4%, 81.8%, and 68.1% for pigs. The SID of AA in 4010 field peas was the least (P < 0.05) for chickens, but in pigs, it was comparable with faba beans. In conclusion, the SID of AA in faba beans and field peas was greater in broiler chickens compared with pigs and there was a cultivar effect.

Faba beans and field peas are legume seeds relatively rich in carbohydrates and protein and may serve as alternative ingredients to reduce dependence on major feed ingredients such as corn and soybean meal. The digestive tract of broiler chickens and pigs is different, and this could influence the utilization of amino acids in feed ingredients. Hence, the objective of this study was to determine and compare the digestibility of amino acids in organic faba beans, and three cultivars of field peas between chickens and pigs. Chickens and pigs were fed diets containing faba beans and field peas as the only source of protein. We determined that 72% to 95% of the essential amino acids in faba beans and field peas were digested by broiler chickens, whereas 68% to 95% were digested by pigs. Also, one cultivar of field peas had lower amino acid digestibility compared with the others. Overall, chickens had greater digestibility of amino acids in these legumes compared with pigs.

Processed fava bean as a substitute for rapeseed meal with or without rumen-protected methionine supplement in grass silage-based dairy cow diets.

Fava bean offers a sustainable home-grown protein source for dairy cows, but fava bean protein is extensively degraded in the rumen and has low Met concentration. We studied the effects of protein supplementation and source on milk production, rumen fermentation, N use, and mammary AA utilization. The treatments were unsupplemented control diet, and isonitrogenously given rapeseed meal (RSM), processed (dehulled, flaked, and heated) fava bean without (TFB) or with rumen-protected (RP) Met (TFB+). All diets consisted of 50% grass silage and 50% cereal-based concentrate including studied protein supplement. The control diet had 15% of crude protein and protein-supplemented diets 18%. Rumen-protected Met in TFB+ corresponded to 15 g/d of Met absorbed in the small intestine. Experimental design was a replicated 4 × 4 Latin square with 3-wk periods. The experiment was conducted using 12 multiparous mid-lactation Nordic Red cows, of which 4 were rumen cannulated. Protein supplementation increased dry matter intake (DMI), and milk (31.9 vs. 30.7 kg/d) and milk component yields. Substituting RSM with TFB or TFB+ decreased DMI and AA intake but increased starch intake. There were no differences in milk yield or composition between RSM diet and TFB diets. Rumen-protected Met did not affect DMI, or milk or milk component yields but increased milk protein concentration in comparison to TFB. There were no differences in rumen fermentation except for increased ammonium-N concentration with the protein-supplemented diets. Nitrogen-use efficiency for milk production was lower for the supplemented diets versus control diet but tended to be greater for TFB and TFB+ versus RSM. Protein supplementation increased plasma essential AA concentration but there were no differences between TFB diets and RSM. Rumen-protected Met clearly increased plasma Met concentration (30.8 vs. 18.2 µmol/L) but did not affect other AA. Absence of differences between RSM and TFB in milk production together with limited effects of RP Met suggest that TFB is a potential alternative protein source for dairy cattle.