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Faba bean ingredients are rich in proteins and good sources of calcium (Ca), although containing phytic acid (PA) molecules. PA, a polyphosphate compound, can affect the bioavailability of minerals/proteins through complex formation. This study evaluates the impact of two extraction processes, Alkaline Extraction-IsoElectric Precipitation (AE-IEP) and Sequential Extraction (SE), on the ability of faba bean globulin systems to bind added calcium ions. Increasing concentrations of CaCl 2 were introduced into 2.5% (w/v) protein dispersions at pHs 4.5, 5.5, 6.5, and 7.5, and free Ca monitored. Near the isoelectric point of globulin (pH ∼ 4-5), Ca binding capacity was found to be low. At higher pHs, significant Ca chelation occurred, initially attributed to free PA binding sites, resulting in the formation of insoluble complexes and subsequent protein precipitation. The AE-IEP globulin fraction exhibited a higher Ca binding capacity than the SE globulin, attributed to its higher PA and lower initial Ca concentrations., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Aluminium (Al) stress is the second-leading abiotic stress on crops. An improved understanding of the response mechanisms of plants to Al stress will provide scientific guidance for enhancing the crops' tolerance to Al stress. In this study, Al stress (50-200 μM AlCl 3 ) caused visible damage to broad bean (Vicia faba L.) roots rather than shoots, which was attributed to Al accumulation and distribution in different tissues. Root transcriptomic analysis revealed that Al stress altered cell wall properties by downregulating lignin synthesis and several xyloglucan endotransglucosylase/hydrolase-, expansin- and peroxidase (POD)-encoding genes, which likely weakened cell extensibility to inhibit root growth. Additionally, Al stress impeded reactive oxygen species scavenging pathways involving POD activity and flavonoid biosynthesis, leading to oxidative damage characterised by malondialdehyde accumulation. These results indicate that optimising cell wall properties and/or enhancing antioxidant processes are crucial for alleviating Al toxicity to broad beans. Interestingly, exogenous application (500 and 1000 μM) of the flavonoid apigenin effectively alleviated Al toxicity in broad bean roots by partially improving the total antioxidant capacity of the roots. This study contributes to understanding the interaction between plants and Al and provides new strategies to alleviate Al toxicity in crops., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

Leaf-cutting ants (Formicidae; Atta spp., Acromyrmex spp.) cut off pieces of leaves and other plant tissue and feed it to their symbiotic fungi. As this foraging behavior poses an imminent threat to agriculture, leaf-cutting ants are considered as pests of huge ecologically and economically importance. Consequently, research on leaf-cutting ants focused on their foraging decisions and interactions with their cultivated symbiotic fungi, whereas their effect on the attacked plants, apart from the loss of plant tissue, remains largely unknown. In this study, we investigated the consequences of an attack by leaf-cutting ants and analyzed the plants' defense responses in comparison to chewing caterpillars and mechanical damage. We found that an attack by leaf-cutting ants induces the production of jasmonates in several host and non-host plant species (Arabidopsis thaliana, Vicia faba, Phaseolus lunatus, Tococa quadrialata). Additionally, we showed in the natural host plant lima bean (P. lunatus) that leaf-cutting ant damage immediately leads to the emission of typical herbivory-induced plant volatiles, including green leaf volatiles and terpenoids. Further data exploration revealed clear differences in the defense-related phytohormone profile in plant species of Neotropical and Eurasian origin. Taken together, we show that leaf-cutting ant infestation and their way of clipping the plants' tissues induce jasmonate and jasmonates-mediated responses and do not differ from those to mechanical injury or larval feeding., (© 2024. The Author(s).)

Reducing the content of quickly absorbed carbohydrates and saturated fats in snack formulations while increasing the consumption of high-quality proteins are effective strategies to prevent obesity in childhood. Thus, the nutritional value, digestibility, and functionality of fava beans (Vicia faba L.) fermented with Pleurotus ostreatus were examined as potential ingredients for food design. Solid-state fermentation enhanced the protein content by 16% with a rise in essential (25%) and non-essential (15%) amino acids while decreasing total carbohydrate content and tannin levels. Moreover, fermentation modified the amino acid profile released during digestion, increasing amino acids such as valine, isoleucine, and threonine, which are vital for health and development in childhood. Furthermore, the bioaccessible fraction of the fermented bean showed a 60% of ACE inhibition and improved magnesium bioaccessibility. Consequently, fava beans fermented with Pleurotus ostreatus emerged as a new ingredient in the development of new protein-rich snacks tailored for children and adolescents., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Mitigating the negative impacts of polycyclic aromatic hydrocarbons (PAHs) is an urgent need due to their toxicity and persistence in the environment. This study investigated the use of Pseudomonas aeruginosa ASU-B6 to detoxify pyrene (PY). The bacterium P. aeruginosa ASU-B6 is capable of degrading PY by 92% as a sole carbon source after 15 days of incubation with phthalate being the major metabolic product. In this regard, the impact of pyrene (PY), P. aeruginosa ASU-B6 (ASU-B6), the bacterial strain combined with pyrene (ASU-B6/PY) and the metabolites produced after pyrene degradation (PY-metabolites) on the germination and physiological attributes of Hordeum vulgare and Vicia faba seedlings were studied. A single application of PY or ASU-B6 showed a toxic effect on the germination of both tested seeds. Interestingly, broad bean seedlings exhibited less sensitivity to PY stress in terms of growth and metabolism compared to barley. Notably, ASU-B6 inhibited fresh and dry weight of shoots and roots of barley and, to a lesser extent, reduced the germination of broad beans compared to the control. However, the combined PY-metabolites and ASU-B6/PY showed a mutual ameliorative effect on seedlings growth, alleviating the phytotoxic impact of each component. Pyrene reduced the virulence of ASU-B6 by inhibiting the production of pyocyanin pigment, while bacteria ameliorated pyrene toxicity through its degradation. Heatmap and principal component analyses highlighted that increasing the contents of hydrogen peroxide, superoxide anion, hydroxyl radical, and lipid peroxidation positively correlated to the toxicity of PY or ASU-B6. However, improving the antioxidant system which buffers the oxidative stress induced by different combinations of PY and ASU-B6 enhanced the growth of germinated seedlings corresponding to PY or ASU-B6. This study reflected the role of ASU-B6 in ameliorating PY-phytotoxicity. In addition, the application of ASU-B6 strain is recommended as a prospective candidate for remediation of PAHs-contaminated environment with a positive impact on the plant growth and metabolic products., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

Enhancement of plant growth at early growth stages is usually associated with the stimulation of various metabolic activities, which is reflected on morphological features and yield quantity and quality. Vitamins is considered as anatural plant metabolites which makes it a safe and ecofriendly treatment when used in appropriate doses, for that this research aimed to study the effect of two different vitamin B forms (thiamine and pyridoxine) on Vicia faba plants as agrowth stimutator in addition to study it's effect on plant as astrong antioxidant under salinity stress.Our findings demonstrated that both vitamin forms significantly increased seedling growth at germination and early growth stages, especially at 50 ppm for pyridoxine and 100 ppm for thiamine. Pyridoxine at 50 ppm increased seedling length by approximately 35% compared to control, while thiamine at 100 ppm significantly promoted seedling fresh and dry wt by 4.36 and 1.36 g, respectively, compared to control seedling fresh wt 2.17 g and dry weight 1.07 g. Irrigation with 100 mM NaCl had a negative impact on plant growth and processes as well as the uptake of several critical ions, such as K + and Mg +2 , increasing Na uptake in comparison to that in control plants. Compared to control plants irrigated with NaCl solution, the photosynthetic pigments, soluble sugars, soluble proteins, and total antioxidant capacity increased in the presence of pyridoxine and thiamine, both at 50 and 100 ppm salinity. The proline content increased in both treated and untreated plants subjected to salt stress compared to that in control plants. Thiamine, especially at 50 ppm, was more effective than pyridoxine at improving plant health under saline conditions. An increase in Vicia faba plant tolerance to salinity was established by enhancing antioxidant capacity via foliar application of vitamin B through direct and indirect scavenging methods, which protect cell macromolecules from damage by oxidative stress, the highest antioxidant capacity value 28.14% was recorded at 50 ppm thiamine under salinity stress.The provided results is aguide for more researches in plant physiology and molecular biology to explain plant response to vitamins application and the suggest the sequence by which vitamins work inside plant cell., (© 2024. The Author(s).)

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., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

In this study, the transepithelial transport of bioactive peptides derived from faba bean flour gastrointestinal digestates was investigated, in vitro, using a Caco-2 and HT29-MTX-E12 coculture monolayer, in comparison to those of pea and soy. The profile of transported peptides was determined by mass spectrometry, and the residual antioxidant activity was assessed. The ORAC value significantly ( p < 0.05) decreased after transepithelial transport (24-36% reduction) for all legumes, while the antioxidant activity in ABTS assay significantly ( p < 0.05) increased, as shown by the EC 50 decrease of 26-44%. Five of the nine faba bean peptides that crossed the intestinal cell monolayer exhibited antioxidant activity. Two of these peptides, TETWNPNHPEL and TETWNPNHPE, were further hydrolyzed by the cells' brush border peptidases to smaller fragments TETWNPNHP and TWNPNHPE. These metabolized peptides were synthesized, and both maintained high antioxidant activity in both ABTS (EC 50 of 1.2 ± 0.2 and 0.4 ± 0.1 mM, respectively) and ORAC (2.5 ± 0.1 and 3.4 ± 0.2 mM of Trolox equivalent/mM, respectively) assays. These results demonstrated for the first time the bioaccessibility of faba bean peptides produced after in vitro gastrointestinal digestion and how their bioactive properties can be modulated during transepithelial transport.

In 2005-2007, a field study was conducted into intercropping of maize with faba bean at Pawlowice research station, Wroclaw University of Environmental and Life Sciences. The main aim of the multi-year field research was an investigation into the reactions of differing maize hybrid earliness to intercropping cultivation with faba bean. The field research evaluated the effect of three maize hybrids-Wilga (early-E), Blask (medium-M) and Iman (late-L)-and the sowing rate of faba bean-18 (Fb1), 27 (Fb2) and 36 (Fb3) seeds per 1 m 2 -on growth dynamics and yield structure, and biomass, protein, and energy yield. Cultivation of faba bean in maize inter-rows led to significant competition with maize and affected yields, causing a decrease in maize dry matter yield from 14.1 (Fb1) to 20.6% (FB3) compared with maize sown alone. In terms of total biomass yield from maize and faba beans, no significant differences were found, but a slight increase in yield of 1.1-4.2% (repective to Fb1 and Fb3) was noted compared to maize sown alone. The early maize hybrid had a significantly lower yield but was most suitable for intercropping with faba bean. The dry biomass yield of early hybrids increased in intercropping by 25% compared to pure maize cultivation. Total protein yield from both intercropping components was higher than in the pure sowing of maize: from 24 (Fb1) to 39% (Fb3). The increase in protein production resulted in an improvement in the energy-protein ratio. The number of UFL per kg of total protein decreased from 13.2 in pure maize cultivation (M-P) to 9.3 (Fb3). A more balanced forage biomass was produced from intercropping maize with faba bean, especially when an early maize hybrid was sown with faba beans., (© 2024. The Author(s).)

Heavy metal pollution threatens human and ecological health. Heavy metals can exist in the soil for a long time and migrate to organisms along the food chain. However, only a few studies have investigated the effects of a single stress on broad beans. Here, we aimed to characterize Cd and Pb bioaccumulation, at varying concentrations, in the broad bean, Vicia faba L. We also determined how the bioaccumulated metals are impacted by aphids that consume the plant. No significant difference was noted in the germination rates of broad beans at the early stage of planting (after 8 days), but eventually, the germination rates of broad beans at all time points first decreased and then increased, and the highest inhibition efficiency was observed in the T3 group (12.5 mg/L Cd 2+  + 50 mg/L Pb 2+ ). Fourteen days after planting, there was no significant difference in seedling height between the T5 (50 mg/L Cd 2+  + 200 mg/L Pb 2+ ) and control groups; however, that in the other groups decreased significantly and there was no dependence between stress concentration and inhibition efficiency. In addition, both Cd and Pb in the soil could be transferred to broad beans, and the concentration of Pb in the roots of broad beans was greater than that of Cd, whereas the opposite was observed in the stems and leaves. Notably, under mixed stress, aphids could significantly reduce the content of Cd in broad beans; similarly, the Pb content in the roots and stems of broad beans decreased significantly after being infested with aphids but increased significantly in the leaves. Further, the aphid infestation decreased the Pb content in the soil and the soil Cd content in the highest concentration group (T5 group) (50 mg/L Cd 2+  + 200 mg/L Pb 2+ ). These results highlight the necessity of focusing on the effect of insects on heavy metal remediation in plants and provide a new perspective for reducing plant Cd toxicity., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

In the course of their life, plants face a multitude of environmental anomaly that affects their growth and production. In recent decades, lead (Pb) gained an increasing attention as it is among the most significant contaminants in the environment. Therefore, in this study the effects of Pb concentrations (0, 50 and 100 ppm) on Vicia faba plants and attempts to alleviate this stress using chitosan (Chs; 0 and 0.1%) were performed. The results validated that with increasing Pb concentrations, a decline in growth, pigments and protein contents was observed. In the same time, a significant upsurge in the stress markers, both malondialdehyde (MDA) and H 2 O 2 , was observed under Pb stress. Nonetheless, foliar spraying with Chs improves the faba bean growth, pigment fractions, protein, carbohydrates, reduces MDA and H 2 O 2 contents and decreases Pb concentrations under Pb stress. Pb mitigation effects by Chs are probably related with the activity of antioxidant enzymes, phenylalanine ammonia lyase (PAL) and proline. The application of Chs enhanced the activities of peroxidase, catalase and PAL by 25.77, 17.71 and 20.07%, respectively at 100 ppm Pb compared to their control. Plant genomic material exhibits significant molecular polymorphism, with an average polymorphism of 91.66% across all primers. To assess the genetic distance created among treatments, the dendrogram was constructed and the results of the similarity index ranged from 0.75 to 0.95, indicating genetic divergence. Our research offers a thorough comprehension of the role of Chs in lessening the oxidative stress, which will encourage the use of Chs in agricultural plant protection., (© 2024. The Author(s).)

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., Competing Interests: No authors have a competing interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Bouraoui et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

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 CuSO 4 ) 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 (H 2 O 2 ) 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., (© 2024. The Author(s).)

Soil acidity is a global issue; soils with pH <4.5 are widespread in Europe. This acidity adversely affects nutrient availability to plants; pH levels <5.0 lead to aluminum (Al 3+ ) toxicity, a significant problem that hinders root growth and nutrient uptake in faba bean (Vicia faba L.) and its symbiotic relationship with Rhizobium. However, little is known about the specific traits and tolerant genotypes among the European faba beans. This study aimed to identify response traits associated with tolerance to root zone acidity and Al 3+ toxicity and potentially tolerant genotypes for future breeding efforts. Germplasm survey was conducted using 165 genotypes in a greenhouse aquaponics system. Data on the root and shoot systems were collected. Subsequently, 12 genotypes were selected for further phenotyping in peat medium, where data on physiological and morphological parameters were recorded along with biochemical responses in four selected genotypes. In the germplasm survey, about 30% of genotypes showed tolerance to acidity and approximately 10% exhibited tolerance to Al 3+ , while 7% showed tolerance to both. The phenotyping experiment indicated diverse morphological and physiological responses among treatments and genotypes. Acid and Al 3+ increased proline concentration. Interaction between genotype and environment was observed for ascorbate peroxidase activity, malondialdehyde, and proline concentrations. Genomic markers associated with acidity and acid+Al 3+ -toxicity tolerances were identified using GWAS analysis. Four faba bean genotypes with varying levels of tolerance to acidity and Al 3+ toxicity were identified., (© 2024 The Author(s). Physiologia Plantarum published by John Wiley & Sons Ltd on behalf of Scandinavian Plant Physiology Society.)

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 15 N balance method., Methods: Nine volunteers completed the study (7 males, 2 females, aged 33 ± 10 y, BMI: 24.7 ± 2.6 kg/m 2 ). 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 15 N. 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., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

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 (EC 50 = 0.5 ± 0.2 mM) and DPPH (EC 50 = 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 (IC 50 = 43 ± 1 μM) and VVIPTEPPHA (IC 50 = 50 ± 5 μM) had the highest activity ( p < 0.05), followed by VIPTEPPH (IC 50 = 90 ± 10 μM) and then VIPTEPPHA (IC 50 = 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.

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., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Fleetwood et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Main Conclusion: The Na + /Ca 2+ 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 Ca 2+ antagonizes Na + toxicity and osmotic stress of NaCl was also targeted in isosmotic combinations of NaCl and CaCl 2 having various Na + :Ca 2+ 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 CaCl 2 (Na + :Ca 2+ 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. Ca 2+ 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., (© 2024. The Author(s).)

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., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Masson SAS. All rights reserved.)

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., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

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., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

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., (© 2023. The Author(s).)

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., (The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

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 (∆AUC 0-180 ), 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.

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.

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., (© 2023. The Author(s).)

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&#95;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 cm 2 and from 74.8 ± 10.6 to 71.5 ± 9.8 cm 2 in the NPN&#95;1 and MPC groups, respectively (P < 0.001). Remobilization partially recovered quadriceps CSA (77.3 ± 9.3 and 72.6 ± 10.0 cm 2 , 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&#95;1 than those with MPC (1.53% ± 0.38% vs. 1.23% ± 0.36%/d, respectively; P = 0.027)., Conclusion: NPN&#95;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&#95;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., (Copyright © 2023 American Society for Nutrition. Published by Elsevier Inc. All rights reserved.)

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., (© 2023. Crown.)

Uranium (U) phytotoxicity is an inherently difficult problem in the phytoremediation of U-contaminated environments. Plant chelating and antioxidant systems play an authoritative role in resistance to abiotic stress. To reveal the toxicity of U, the changes of chelating system, osmoregulatory substances and antioxidant systems in Vicia faba roots were studied after short-term (24 h) U exposure. The results indicated that the development of lateral roots and root activity of V. faba were significantly inhibited with U accumulation. Compared with the control, plant chelating systems showed significant positive effects after U exposure (15 - 25 μM). Osmoregulatory substances (proline and soluble protein) increasingly accumulated in roots with increasing U concentration, and O 2 - and H 2 O 2 rapidly accumulated after U exposure (15 - 25 μM). Thus, the contents of malondialdehyde (MDA), a marker of lipid peroxidation, were also significantly increased. Antioxidant systems were activated after U exposure but were inhibited at higher U concentrations (15 - 25 μM). In summary, although the chelating, osmotic regulation and antioxidant systems in V. faba were activated after short-term U exposure, the antioxidases (CAT, SOD and POD) were inhibited at higher U concentrations (15 - 25 μM). Therefore, the root cells were severely damaged by peroxidation, which eventually resulted in inhibited activity and arrested root development., (© 2023. The Author(s) under exclusive licence to The Botanical Society of Japan.)

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., (The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Recent studies have shown that Cr uses other element transporters such as phosphate transporters to enter cells. The aim of this work is to explore the interaction between dichromate and inorganic phosphate (Pi) in the plant of Vicia faba L. To study this interaction, we used three concentrations of Dipotassium hydrogen phosphate (K 2 HPO 4 ) 10 mM (Pi10), 50 mM (Pi50) and 100 mM (Pi100) added alone or in combination with potassium dichromate (K 2 Cr 2 O 7 ) Cr + Pi10, Cr + Pi50 and Cr + Pi100. In order to investigate the impact of this interaction on morpho-physiological parameters, the biomass, chlorophyll content, proline level, H 2 O 2 level, Catalase and Ascorbate peroxidase activity and Cr-bioaccumulation has been determined. For the molecular scale, the theoretical chemistry was used via molecular docking to explore the various interactions between dichromate Cr 2 O 7 2- /HPO 4 2- /H 2 O 4 P - and the phosphate-transporter. We have selected the eukaryotic phosphate transporter (PDB: 7SP5) as the module. The results showed that K 2 Cr 2 O 7 negatively affects morpho-physiological parameters and generates oxidative damage (+84% H 2 O 2 than the control), which involved the production of antioxidant enzymes (+147% Catalase and +176% Ascorbate-peroxidase) and Proline (+108%). The addition of Pi improved the growth of Vicia faba L. and induces the partial restoration of the parameters affected by Cr (VI) to the normal levels. Also, it decreased oxidative damage and reduce Cr (VI) bioaccumulation in shoots and roots. Molecular docking has shown that the dichromate structure is more compatible and establishes more bonds with the Pi-transporter which generates a very stable complex compared to HPO 4 2- /H 2 O 4 P - . Overall, these results confirmed that there is a strong relationship between dichromate uptake and the Pi-transporter., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Masson SAS. All rights reserved.)

Increasing the proportion of locally produced plant protein in currently meat-rich diets could substantially reduce greenhouse gas emissions and loss of biodiversity 1 . However, plant protein production is hampered by the lack of a cool-season legume equivalent to soybean in agronomic value 2 . Faba bean (Vicia faba L.) has a high yield potential and is well suited for cultivation in temperate regions, but genomic resources are scarce. Here, we report a high-quality chromosome-scale assembly of the faba bean genome and show that it has expanded to a massive 13 Gb in size through an imbalance between the rates of amplification and elimination of retrotransposons and satellite repeats. Genes and recombination events are evenly dispersed across chromosomes and the gene space is remarkably compact considering the genome size, although with substantial copy number variation driven by tandem duplication. Demonstrating practical application of the genome sequence, we develop a targeted genotyping assay and use high-resolution genome-wide association analysis to dissect the genetic basis of seed size and hilum colour. The resources presented constitute a genomics-based breeding platform for faba bean, enabling breeders and geneticists to accelerate the improvement of sustainable protein production across the Mediterranean, subtropical and northern temperate agroecological zones., (© 2023. The Author(s).)

Background: Assessment of protein quality is necessary to satisfy the nutritional needs of populations across the world. In addition to indispensable amino acid (IAAs) composition, protein digestibility is a major component of IAA bioavailability, playing a crucial role in human health and affecting the linear growth of children., Objectives: This study aimed to evaluate IAA digestibility of fava beans, a legume widely consumed in Morocco using the dual-tracer method., Methods: 2 H-intrinsically labeled Fava beans supplemented with 12 mg/kg BW of 13 C spirulina were given to 5 healthy volunteers (3 men and 2 women), aged 25.8 ± 3.3 y, with a mean BMI of 20.0 kg/m 2 . The meal was spread in small portions and was given hourly throughout 7 h. Blood was sampled at baseline and hourly from 5 to 8 h after meal ingestion. IAA digestibility was evaluated by gas chromatography-combustion-isotope ratio mass spectrometry using the 2 H/ 13 C ratio in plasma IAA. Digestible indispensable amino acid ratios (DIAAR) were calculated using the scoring pattern for people older than 3 y., Results: Fava beans had an adequate level of lysine but were limiting in several IAAs, especially methionine. Under our experimental conditions, the average IAA digestibility of fava bean was 61.1% ± 5.2%. Valine had the highest digestibility (68.9% ± 4.3%) and threonine had the lowest (43.7% ± 8.2%). In consequence, the lowest DIAAR was 67% for threonine and only 47% for sulfur amino acids (SAA)., Conclusions: The present study is the first to determine the digestibility of fava bean amino acids in humans. The mean IAA digestibility was moderate, and consequently, we conclude that fava bean provides a limited amount of several IAAs, especially SAA, but adequately for lysine. Preparation and cooking methods of fava beans should be improved to increase digestibility. This study was registered at ClinicalTrials.gov as NCT04866927., (Copyright © 2023 American Society for Nutrition. All rights reserved.)

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 ± 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 ± 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., (© The Author(s) 2023. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

The objective of this study was to investigate the effects of heat treatment (toasting) and particle size alterations (grinding; rolling) on nutrient utilization, ruminal fermentation, and supply of metabolizable protein (MP), and to study the interaction between processing conditions of fava beans and forage type. Six Danish Holstein dairy cows fitted with ruminal, duodenal, and ileal cannulas were used in a 6 × 4 incomplete Latin square design with 4 periods of 21-d duration. Cows were fed ad libitum with 6 experimental diets: diets high in either grass-clover silage or corn silage were combined with ground untoasted, ground toasted, or rolled untoasted fava beans. Samples of ruminal fluid, digesta from duodenum and ileum, and feces were collected, and nutrient digestibility was estimated using Cr 2 O 3 and TiO 2 as flow markers. Diets high in corn silage resulted in higher ruminal pH and higher proportion of propionate in ruminal volatile fatty acids compared with diets high in grass-clover silage. Diets high in corn silage resulted in higher apparent total-tract digestibility of crude protein and starch but lower apparent ruminal and total-tract digestibility of neutral detergent fiber compared with diets high in grass-clover silage. Rolling of fava beans decreased the in situ small intestinal disappearance of rumen-undegradable protein corrected for particle losses. Compared with grinding, rolling of fava beans reduced apparent ruminal digestibility of starch, true ruminal digestibility of organic matter, crude protein, and AA, and small intestinal digestibility of AA and starch. Grinding of fava beans increased apparent ruminal digestibility of neutral detergent fiber and reduced the proportion of propionate in ruminal volatile fatty acids compared with rolling of fava beans. In addition, rolling of fava beans had no effect on MP supply. Toasting of fava beans had no effect on in vivo nutrient digestibility except for an interaction with forage source on apparent ruminal dry matter and organic matter digestibility. Toasting of fava beans did not affect small intestinal digestion of individual and total AA, and therefore failed to increase MP supply. In conclusion, neither replacing grass-clover silage with corn silage, nor toasting nor rolling of fava beans had an effect on supply of MP., (The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

In order to further explain the fillet texture improvement of Yellow River carp (Cyprinus carpio) fed with faba bean (Vicia faba L.), a three-month rearing trial was conducted to investigate fatty acid composition, antioxidant capacity, myofiber development, collagen deposition and transcriptome in white muscle of two farmed carp groups (One was fed only faba bean, the other was fed commercial diet). As a strong oxidant, faba bean changed fatty acids composition in white muscle, especially DHA and EPA, up-regulated the levels of reactive oxygen species (ROS) and down-regulated major antioxidant enzyme activities in the hepatopancreas and white muscle. Through the analysis of transcriptome and subsequent verification analysis, we speculated that the increase of ROS led to the decrease of myofiber diameter and collagen metabolism. This study provides a theoretical basis for further understanding the regulation of faba bean on fillet texture characteristic of Yellow River carp., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Lupines and faba beans are promising ingredients for the beverage industry. They contain high amounts of protein and can be grown in different climate zones and agricultural areas. Therefore, these legumes appear as ideal raw material for vegan, functional, and sustainable beverages. Nevertheless, the sensory characteristic of legumes is generally not accepted in beverages. Therefore, the market contribution of legume-based beverages is currently only marginal. This review highlights known major flavor aspects of lupines and faba beans and the possibilities to improve these by germination, heat treatment, enzymatic treatment, and subsequent lactic acid fermentation. First, the main aroma and taste compounds are described. Thereby, the "beany" aroma is identified as the most relevant off-flavor. Second, the nutrients and antinutrients of these legumes regarding to their use as food and as substrate for lactic acid fermentation are reviewed, and possibilities to modulate the substrate are summarized. Finally, the modification of the sensory profile by lactic acid fermentation is outlined. To conclude, it seems likely that the nutritional and flavor attributes in legume-based beverages can be improved by a combined process of substrate modulation and fermentation. In a first step, antinutrients should be decomposed and proteins solubilized while transforming the solid grains into a liquid substrate. Due to such substrate modulation, a broader variety of strains could be employed and the fermentation could be based exclusively on their impact on the flavor. By applying the concept of combining a substrate modulation with a subsequent fermentation, the use of legumes in beverages could be facilitated and new products like vegan, protein-rich, refreshing beverages could be marketed., (© 2022 The Authors. Comprehensive Reviews in Food Science and Food Safety published by Wiley Periodicals LLC on behalf of Institute of Food Technologists.)

Meat analogs based on plant protein extracts are rising in popularity as meat consumption declines. A dietary shift away from meat, which has a high iron bioavailability, may have a negative effect on the amount of iron absorbed from the diet. Iron absorption from legumes cultivated in regions not suitable for soy production, such as fava bean, has not yet been explored. The aim of this study was to evaluate non-heme iron absorption from a meal with texturized fava bean protein compared to beef and cod protein meals. The study included two single-blinded iron isotope trials in healthy Swedish women of the ages 18-45 years, each of whom served as their own control. The participants were served matched test meals containing beef and fava bean protein (Study 1) or cod and fava bean protein (Study 2) with radiolabeled non-heme iron 55 Fe and 59 Fe. The absorption of non-heme iron from test meals was measured by whole-body counting and erythrocyte incorporation. The absorption of non-heme iron, measured as erythrocyte incorporation ratio, from beef protein meal was 4.2 times higher compared to texturized fava bean meal, and absorption from cod protein meal was 2.7 times higher compared to the fava bean meal. The adjusted non-heme iron absorption, normalized to a 40% reference dose uptake, was 9.2% for cod protein meal, 21.7% for beef protein meal, and 4.2% for texturized fava bean meal. A fava bean protein meal has markedly lower iron bioavailability in healthy females compared with a meal of beef or cod protein. Therefore, a dietary shift from meat and fish protein to fava bean protein may increase the risk of iron deficiency.

Rhizosphere soil microbial community and its diversity are important for the structure and functions of medicinal plant ecosystem. In this study, based on high-throughput sequencing, rhizosphere soil microbial diversity, and yield and quality of rhizome and root of Notopterygium incisum cultivated alone(control, CK) and intercropped with Vicia faba(QH) were analyzed, which is expected to lay a basis for optimization of the cultivation mode and ecological production of N. incisum. RESULTS:: showed that the rhizosphere soil bacteria of N. incisum were dominated by Proteobacteria and Bacteroides, with the relative abundance of 50.38%-51.95% and 16.36%-17.02%, respectively. Soil bacterial community at the phylum level was not significantly different between CK and QH. At the genus level, the relative abundance of MND1(3.54%), Spinstomonas(3.50%), Nitrospira(1.53%), and Rhizobacter(1.05%) was significantly higher and that of Gemmatimonas, Candidatus＿Solibacter, and Bryophytes was lower in QH treatment than in the CK. The plant height, leaf length, leaf width, and petiole length of N. incisum in QH treatment was significantly increased and the underground biomass rose by 71.43% compared with those in the CK. Thus, intercropping with V. faba promoted the aboveground growth of N. incisum and improved the yield of root and rhizome. Moreover, the content of notopterol and isoimperatorin increased by 37.96% and 4.09% in QH treatment, respectively, indicating that the intercropping with V. faba boosted the accumulation of secondary metabolites in N. incisum. Pearson's correlation analysis showed that the soil bacterial community was mainly influenced by the soil factors including the content of soil available nutrients, soil organic matter, pH value, and soil water. The influence was in the order: total potassium&gt;total nitrogen&gt;pH&gt;organic matter&gt;available potassium&gt;soil water content&gt;available nitrogen&gt;available phosphorus. In conclusion, the intercropping with V. faba altered soil microenvironment and also increased the yield and accumulation of secondary metabolites of N. incisum, which is a promising ecological planting model for N. incisum.

Background: The variation in amino acid (AA) digestibility and metabolisable energy (ME N ) in four spring and four winter faba bean genotypes differing in vicine/convicine (V/C) concentrations grown on two sites was investigated in caecectomised LSL-Classic laying hens. Effects of dehulling one faba bean genotype were also examined. Diets containing one out of 17 faba bean variants each and a basal diet were fed to ten caecectomised laying hens in a row-column design to achieve five replicates per diet., Results: Ranges and levels of digestibility of the hulled variants differed widely among AA with the lowest and highest range determined for Arg (90-93%) and Cys (-12-65%), respectively. ME N ranged between 10.3 and 12.3 MJ kg -1 dry matter. Lower ME N and digestibility of Cys, Glx, Phe, Pro, Tyr, and Val (P < 0.050) was determined for the winter genotypes grown in Nimtitz compared to the other variants. Digestibility of Ser was lower for the spring than for the winter genotypes (P < 0.050). Negative correlations with AA digestibility were determined for phytate, but not for tannin and V/C concentrations (P < 0.050). Negative correlations between tannin fractions and ME N were weak (P = 0.082-0.099). Dehulling increased ME N by 1.8 MJ kg -1 dry matter and raised the digestibility of Pro, His, and Glx (P < 0.050)., Conclusions: The results indicated that the digestible AA and ME N supply of laying hens was increased by using low phytate faba beans while breeding for low V/C genotypes did not affect AA digestibility or ME N . Dehulling increased ME N and the digestibility of some AA. © 2021 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry., (© 2021 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.)

Rationale: Favism is a well-known cause of acute hemolytic anemia. Rarely, methemoglobinemia can also happen because of fava bean ingestion in patients with glucose-6-phosphate dehydrogenase (G6PD) deficiency. Few cases with this co-occurrence have been reported in the literature., Patient Concerns: We report a case of a 47-year-old patient who presented with jaundice that started 2 days after eating fava beans., Diagnoses: Laboratory investigations revealed anemia with evidence of hemolysis (high reticulocytes count, high indirect bilirubin, bite cells in peripheral smear). Blood gases showed high methemoglobin level. Reduced level of G6PD enzyme confirmed the diagnosis of G6PD deficiency., Intervention: The patient was kept on supplemental oxygen. He was counselled to avoid food and drugs that can cause acute hemolysis., Outcomes: Oxygen saturation improved gradually. The patient was discharged without any complications after 2 days., Lessons: Patients with G6PD deficiency can develop both acute hemolytic anemia and methemoglobinemia secondary to fava beans ingestion. These patients should not receive methylene blue to avoid worsening hemolysis., Competing Interests: The authors have no conflicts of interest to disclose., (Copyright © 2021 the Author(s). Published by Wolters Kluwer Health, Inc.)

Faba bean (Vicia faba L.), a globally important grain legume providing a stable source of dietary protein, was one of the earliest plant cytogenetic models. However, the lack of draft genome annotations and unclear structural information on mRNA transcripts have impeded its genetic improvement. To address this, we sequenced faba bean leaf transcriptome using the PacBio single-molecule long-read isoform sequencing platform. We identified 28,569 nonredundant unigenes, ranging from 108 to 9669 bp, with a total length of 94.5 Mb. Many unigenes (3597, 12.5%) had 2-20 isoforms, indicating a highly complex transcriptome. Approximately 96.5% of the unigenes matched sequences in public databases. The predicted proteins and transcription factors included NB-ARC, Myb&#95;domain, C3H, bHLH, and heat shock proteins, implying that this genome has an abundance of stress resistance genes. To validate our results, we selected WCOR413-15785, DHN2-12403, DHN2-14197, DHN2-14797, COR15-14478, and HVA22-15 unigenes from the ICE-CBF-COR pathway to analyze their expression patterns in cold-treated samples via qRT-PCR. The expression of dehydrin-related genes was induced by cold stress. The assembled data provide the first insights into the deep sequencing of full-length RNA from faba bean at the single-molecule level. This study provides an important foundation to improve gene modeling and protein prediction., (© 2021. The Author(s).)

Background: Esters are indispensable aroma compounds and contribute significantly to the fruity aromas in fermented condiments. The ester synthesis activity and pathways of Bacillus licheniformis, Candida etchellsii, and Zygosaccharomyces rouxii, isolated from Chinese horse bean chili-paste (CHCP), were investigated. Chemical buffer models containing esterification and alcoholysis systems inoculated with extracellular extracts of these three strains were established., Results: The ester synthesis activity of C. etchellsii was stronger than that of the other two strains. Zygosaccharomyces rouxii could synthesize acetate esters via esterification, whereas the biosynthesis pathways of B. licheniformis and C. etchellsii were esterification and alcoholysis. Esterification exhibited relatively high activity at pH 4, whereas alcoholysis activity improved with an increase in the pH from 4 to 8. Candida etchellsii could synthesize C 6 -C 8 of acetate esters, and its activity improved with the number of alcohol carbon atoms. These three strains could synthesize C 10 -C 18 of ethyl esters. Their ethyl ester synthesis activity decreased with the aliphatic acid carbon number., Conclusion: Candida etchellsii has the potential to be used in CHCP fermentation to accumulate esters and improve flavor compared with the other two strains. This research is helpful in explaining the mechanism of ester synthesis in fermented condiments. © 2021 Society of Chemical Industry., (© 2021 Society of Chemical Industry.)

Suitability of poor quality water for irrigation depends on salinity level and solute concentration in the water and selected crop. Salt stress is a major potential constraint for faba bean. The present study aimed to investigate the effects of different Cl- and SO 4 -containing salt sources in irrigation water with different salinity levels on emergence, early seedling growth and photosynthetic capacity of six faba bean genotypes. The negative effect order of salinity level was high (3 dS/m) > medium (2 dS/m) > low (1 dS/m) > control (0.05 dS/m) for all investigated parameters except dry root weight. The negative effects of Cl-containing salt sources were higher than that of SO 4 -containing salt sources. The worst and the best performing genotypes were determined as III-28 and III-29 on emergence percentage at 10th DAS, I-29 and III-1 on mean emergence time, III-22 and III-1 on shoot height, III-1 and I-29 on fresh biomass weight, III-22 and III-28 on fresh shoot weight, III-29 and I-29 on fresh root weight, respectively. This study showed that faba bean genotypes have different behaviors in terms of response to the increasing salinity levels artificially makeup by using different salt sources indicating that salt response of faba bean is genotype-specific., (© 2021. The Author(s).)

This study investigated alkaline shifting, ultrasonication, and their combination on functional modifications of faba bean protein isolate (FPI). The protein characterization revealed that the combined alkaline shifting/ultrasound treatment dissociated large aggregates of FPI into small ones, resulting in increased protein surface hydrophobicity and decreased free sulfhydryl groups. Such structural changes significantly increased FPI solubility from 12.2 to 40.4% to more than 95% at pH 3 and 7 and more than 80% at pH 6. Furthermore, the FPI modified by the combined alkaline shifting/ultrasound treatment produced emulsions with smaller particle size and superior stability at pH values 3 and 7. Likewise, a striking enhancement in foaming capacity (from 93% to 306-386%) and stability (from 10 s to 473-974 s) was achieved by the combined treatment. This study suggested that the ultrasound-assisted alkaline shifting treatment could be considered an effective method for improving FPI functional properties., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Key Message: Faba bean genotypes showed significant and marked genetic differences in their success as pollen donors to cross-fertilized seeds. The findings may improve exploitation of heterosis in synthetic cultivars. In partially allogamous crops such as faba bean (Vicia faba L.), increasing the share of heterosis in a synthetic cultivar can improve yield and yield stability. The share of heterosis in such synthetic cultivars is increased by higher degrees of cross-fertilization. This trait is defined as percentage of cross-fertilized seeds among all seeds and is a crucial parameter in breeders' yield predictions. Current approaches use degree of cross-fertilization to predict inbreeding and share of heterosis, they even consider genotype-specific degrees; yet, all genotypes are assumed to contribute equally to the cross-fertilized seeds. Here, we expect faba bean genotypes to differ in their success rates as pollen donors, i.e. in paternal outcrossing success. To quantify the variation of both, the degree of cross-fertilization and the paternal outcrossing success, we assessed these parameters in inbred lines and F1 hybrids, grown in four polycrosses composed of eight genotypes each. We identified the paternal genotype of 500 to 800 seeds per genotype and polycross using SNP markers. In both traits, we found marked and significant variation among inbred lines and among F1 hybrids, as well as between inbred lines and F1. Based on our findings, we discuss how differential paternal outcrossing success influences the amount of inbreeding in synthetic cultivars. Our findings offer the potential for a better management and exploitation of heterotic yield increase in faba bean., (© 2021. The Author(s).)

Although Australia is the largest exporter of faba bean globally, there is limited information available on the levels of bioactive compounds found in current commercial faba bean varieties grown in this country. This study profiled the phenolic acid and flavonoid composition of 10 Australian faba bean varieties, grown at two different locations. Phenolic profiling by HPLC-DAD revealed the most abundant flavonoid to be catechin, followed by rutin. For the phenolic acids, syringic acid was found in high concentrations (72.4-122.5 mg/kg), while protocatechuic, vanillic, p -hydroxybenzoic, chlorogenic, p -coumaric, and trans-ferulic acid were all found in low concentrations. The content of most individual phenolics varied significantly with the variety, while some effect of the growing location was also observed. This information could be used by food processors and plant breeders to maximise the potential health benefits of Australian-grown faba bean.

Faba bean (Vicia faba L.) is a widely adapted and high-yielding legume cultivated for its protein-rich seeds 1 . However, the seeds accumulate the pyrimidine glucosides vicine and convicine, which can cause haemolytic anaemia (favism) in 400 million genetically predisposed individuals 2 . Here, we use gene-to-metabolite correlations, gene mapping and genetic complementation to identify VC1 as a key enzyme in vicine and convicine biosynthesis. We demonstrate that VC1 has GTP cyclohydrolase II activity and that the purine GTP is a precursor of both vicine and convicine. Finally, we show that cultivars with low vicine and convicine levels carry an inactivating insertion in the coding sequence of VC1. Our results reveal an unexpected, purine rather than pyrimidine, biosynthetic origin for vicine and convicine and pave the way for the development of faba bean cultivars that are free of these anti-nutrients., (© 2021. The Author(s), under exclusive licence to Springer Nature Limited.)

The dry matter partitioning is the product of the flow of assimilates from the source organs (leaves and stems) along the transport route to the storage organs (grains). A 2-year field experiment was conducted at the agronomy research farm of the University of Agriculture Peshawar, Pakistan during 2015-2016 (Y1) to 2016-2017 (Y2) having semiarid climate. Four summer crops, pearl millet (Pennisetum typhoidum L.), sorghum (Sorghum bicolor L.) and mungbean (Vigna radiata L.) and pigeonpea (Cajanus cajan L.) and four winter crops, wheat (Triticum aestivum L.), barley (Hordeum vulgare L.), fababean (Vicia faba) and rapeseed (Brassica napus) were grown under two irrigation regimes (full vs. limited irrigation) with the pattern of growing each crop either alone as sole crop or in combination of two crops in each intercropping system under both winter and summer seasons. The result showed that under full irrigated condition (no water stress), all crops had higher crop growth rate (CGR), leaf dry weight (LDW), stem dry weight (SDW), and spike/head dry weight (S/H/PDW) at both anthesis and physiological maturity (PM) than limited irrigated condition (water stress). In winter crops, both wheat and barley grown as sole crop or intercropped with fababean produced maximum CGR, LDW, SDW, S/H/PDW than other intercrops. Among summer crops, sorghum intercropped either with pigeon pea or with mungbean produced maximum CGR, LDW, SDW, and S/H/PDW at both growth stages. Sole mungbean and pigeon pea or pigeon pea and mungbean intercropping had higher CGR, LDW, SDW, S/H/PDW than millet and sorghum intercropping. On the other hand, wheat and barley grown as sole crops or intercropped with fababean produced maximum CGR, LDW, SDW, and S/H/PDW than other intercrops. Fababean grown as sole crop or intercropped with wheat produced higher CGR, LDW, SDW, and S/H/PDW at PM than intercropped with barley or rapeseed. From the results it was concluded that cereal plus legume intercropping particularly wheat/fababean in winter and sorghum/pigeon pea or sorgum/mungbean in summer are the most productive intercropping systems under both low and high moisture regimes.

Key Message: Modified pEAQ-HT-DEST1 vectors were used for agroinfiltration in legumes. We demonstrate protein expression and export in pea, lentil, and faba bean; however, the method for chickpea was not successful. Agroinfiltration is a valuable research method for investigating virulence and avirulence effector proteins from pathogens and pests, where heterologous effector proteins are transiently expressed in plant leaves and hypersensitive necrosis responses and other effector functions can be assessed. Nicotiana benthamiana is widely used for agroinfiltration and the characterisation of broad-spectrum effectors. The method has also been used in other plant species including field pea, but not yet developed for chickpea, lentil, or faba bean. Here, we have modified the pEAQ-HT-DEST1 vector for expression of 6 × histidine-tagged green-fluorescent protein (GFP) and the known necrosis-inducing broad-spectrum effector necrosis and ethylene-inducing peptide (Nep1)-like protein (NLP). Modified pEAQ-based vectors were adapted to encode signal peptide sequences for apoplast targeting of expressed proteins. We used confocal microscopy to assess the level of GFP expression in agroinfiltrated leaves. While at 3 days after infiltration in N. benthamiana, GFP was expressed at a relatively high level, expression in field pea and faba bean at the same time point was relatively low. In lentil, an expression level of GFP similar to field pea and faba bean at 3 days was only observed after 5 days. Chickpea leaf cells were transformed at low frequency and agroinfiltration was concluded to not be successful for chickpea. We concluded that the pEAQ vector is suitable for testing host-specific effectors in field pea, lentil, and faba bean, but low transformation efficiency limits the utility of the method for chickpea.