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Background: This study investigated the effects of the different concentrations of CaCl 2 (10 and 15 mol m -3 ) on the growth, physiology, and cytological characteristics of salt-stressed Vicia faba (L.) seedlings grown under greenhouse conditions., Results: Salinity stress (150 mol m -3 NaCl) had detrimental effects on all measured growth parameters, increased the micronucleus count number (MCN) by 26.6 micronuclei/1000 cells, decreased the mitotic index (MI) by 66.6%, and caused various chromosomal aberrations, nuclear alterations, and chromatin bridges in salt-stressed seedlings compared to the untreated plant. Nevertheless, the seed priming with CaCl 2 (10 and 15 mol m -3 ) significantly alleviated the toxic effects induced by salinity stress, improved growth parameters, total chlorophyll (TChl), proline, and total soluble sugar (TSS) contents in salt-stressed faba bean seedling compared with seedlings germinated from non-primed seeds. The antioxidative system of salt-stressed faba bean was highly stimulated by increasing the activity of catalase (CAT), peroxidase (POD), and superoxide dismutase (SOD) enzymes as well as phenolics and flavonoids were increased in all salt-stressed seedlings germinated from seeds primed with CaCl 2 (10 and 15 mol m -3 ) indicating an improved tolerance of faba bean plant to salinity stress. Notably, the pretreatment with CaCl 2 (10 mol m -3 ) reduced the micronuclei number per 1000 cells by 91.3% and decreased the abnormality index by 58.9% more effectively than CaCl 2 (15 mol m -3 ). SDS-PAGE profiling revealed the presence of 16 proteins with different molecular weights, including two peptides, induced by CaCl 2 (10 mol m -3 ) in response to salinity stress., Conclusions: This study showed that 10 mol m -3 CaCl 2 significantly improved salt tolerance in treated faba bean plants mitigating the antagonistic effects of salt stress on several physiological and cytological parameters., Competing Interests: Declarations. Ethics approval and consent to participate: Not applicable. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

Background: One of the primary challenges that the expanding population faces is water scarcity. Thus, a global imperative has been established to safeguard extant water resources and optimize their utility through sustainable practices and efficient management. In the present investigation, Azolla pinnata, a pteridophyte (fern), was employed to phytoremediate Cr (VI) from chromium-polluted water. The potential of this treated water for agricultural purposes was verified through the use of Vicia faba plants., Results: In vitro, A. pinnata effectively remediates Cr (VI) from an array of liquid concentrations (0.05 to 90 ppm) in a ratio of 25:1 {volume (mL): fresh weight of Azolla (g)} after 2 days incubation period at room temperature. At low concentrations (0.1 ppm), the phytoremediation capacity peaked at 70%, falling to 19.53% at a high concentration (90 ppm). Upon continuous irrigation with Cr-polluted water (0.05 to 50 ppm), the in vivo pot experiment on Vicia faba plants revealed high Cr accumulation in the roots reached 52.5 mg Kg -1 dry weight (Dwt) at the 50 ppm Cr treatment. Nevertheless, a reduced Cr content of 19.5 mg Kg -1 Dwt was observed when the plants were irrigated with 50 ppm Cr-polluted water that had been treated with Azolla. At 50 ppm of Cr, Azolla's treatment significantly increased shoot length, fresh weight, and Chl a content to 25.25 cm, 3.4 g, and 6.5 mg g -1 Dwt, respectively, up from 10.25, 1.8, and 4.7 in untreated plants. The chromosomal aberrations were significantly induced in the dividing cells of all Cr treatments, with the highest value of 4.8% at 50 ppm. This value was reduced to 2.88% at the same concentration when treated with Azolla. At a concentration of 10 ppm Cr, the mitotic index was significantly improved to 6.99% when combined with Azolla, as opposed to 3.63% when the same concentration was used without Azolla. The DNA degradation assay showed partial DNA degradation at 50 ppm Cr, which the Azolla treatment eliminated. Furthermore, the gene expression levels of both the PM H + -ATPase and the calcium-dependent protein kinase CDPK5 were upregulated in response to Cr, despite the fact that the expression level was altered in a dose- and concentration-dependent manner by Azolla treatment., Conclusion: Azolla exhibits substantial potential for reducing the detrimental effects of chromium stress including oxidative stress on plants. It modulates stress-related gene expression, protects DNA integrity, enhances cell mitosis, and reduces chromosomal damage. These results indicate that Azolla has the potential to be a valuable asset in phytoremediation strategies for chromium-contaminated environments, and that it may enhance plant survival and growth under Cr stress conditions., Key Message: Azolla pinnata can be effectively utilized as an environmentally-friendly method to remediate chromium-contaminated water for agricultural usage., Competing Interests: Declarations. Experimental research and field studies on plants: “All relevant institutional, national and international guidelines and legislation were compiled or adhered to the production of this study”. Ethics approval and consent to participate: This article does not contain any studies with human participants or animals performed by any of the authors. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

The current study aimed to detect the mutagenic impacts of aflatoxin B1 (AFB1), which is produced by Aspergillus group fungi, via a high-plant genotoxicity test. Different durations of treatment (3 h, 6 h, and 12 h) were used to treat the Vicia faba root tips with varying concentrations of Aflatoxin B1 (AFB1) following the approved protocol for plant assays published by the International Program on Chemical Safety (IPCS) and the World Health Organization (WHO). The data obtained indicated that AFB1 not only has the ability to induce various alterations in the process of mitosis, ranging from increasing to decreasing mitotic and phase indices but also leads to many mitotic aberrations. The abnormalities observed varied on the basis of the ratio of AFB1 to treatment time. The aberrations included micronuclei in interphase, stickiness; two groups ring star disturbed and oblique metaphase late separation diagonal bridge and laggard and disturbed. anaphase and telophase. This study showed that biomonitoring Vicia faba is a sustainable method for estimating the cytotoxicity and genotoxicity of applied AFB1. Additionally, AFB1 caused changes in the protein profile detected by SDS‒PAGE, with each treated sample displaying a unique electrophoretic pattern due to the formation and disappearance of certain bands. The ISSR and RAPD assays changes in band numbers in all samples compared with the untreated control, and a decrease in genetic template stability (GTS) ratios was observed with higher levels of AFB1. The image cytometric data revealed a correlation between the dosage of AFB1 and its impact on cell cycle components in the meristematic cells of Vicia faba roots. Furthermore, an increase in AFB1 concentrationled to a decrease in B-cell lymphoma 2 (Bcl2) levels, an increase in chromatin condensation levels, and an increase in poly ADP‒ribose polymorphism (PARP) levels., Competing Interests: Declarations. Ethics approval and consent to participate: Not applicable. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

Faba bean is a nutritionally and medicinally rich popular legume crop. However, vicine-convicine remain as potential threats for "favism" in human beings. In this study, 189 diverse faba bean accessions have been evaluated for yield component traits and vicine content in seeds followed by a correlation study. Combined genetic variability analysis shows that traits like days to pod initiation (DPI), pod length (PL), test weight (TW) and grain yield have minimally been influenced by the environment. PCA revealed that TW, PL and PW were the primary indicators for deciding yield performance. LC-MS/MS confirms that vicine concentration varied in between 3.489 and 10.025 g/kg and a significant positive correlation (0.40***) was observed between vicine conc. and grain yield of faba bean. Thus, present study demonstrated that the faba bean genotypes containing lower vicine were mostly poor yielding, which might be regulated by vicine in faba bean. Therefore, complete elimination of vicine or development of near-zero vicine faba bean could drastically reduce the yield potential of the crop, hence one has to be very cautious and follow efficient selection strategies while optimizing lower concentration of vicine for development of low vicine varieties. This study shows that faba bean genotypes containing 4.0-5.5 g/kg vicine were fairly productive and also have considerably lower vicine., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Faba bean is an important legume crop with significant potential to contribute to sustainable agricultural systems and food security in Ethiopia. Despite its importance, the crop is prone to various biotic and abiotic constraints that can reduce seed yield and affect its stability and adaptability. To identify stable and adaptable genotypes, 10 faba bean genotypes were evaluated at three locations over two growing seasons using different stability parameters. Genotype-by-environment interaction (GGE biplot) and additive main effect and multiplicative interaction (AMMI) analyses are the statistical methods used to evaluate crop genotype performance across different environments to identify high-performing, stable, and adaptable genotypes and to highlight preferable environments for genotype differentiation. This study utilized cultivar superiority, regression coefficients, and deviations from regression parameters that provide valuable insights into genotype performance under varying environmental conditions. This approach helps to identify robust cultivars that can thrive across different agricultural settings and challenges, ultimately contributing to improved crop production and food security. The results revealed that G9, G8, and G7 are the three most stable and adaptable genotypes. These faba bean genotypes showed greater resilience to environmental changes and improved suitability for sustainable production, making them better options for local farmers. They also bolster resilience against climate variability and ultimately ensure agricultural sustainability. The AMMI model indicated that the genotype-environment interaction (GEI) significantly influences seed yield. These findings provide crucial insights into the genetic potential of faba bean genotypes that can help breeding programs to develop high-yielding, adaptable, and stable varieties for the region and other areas with similar agro-ecological conditions., Competing Interests: Declarations. Ethics approval and consent to participate: Not applicable. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

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.)

Background: Plant mutagenesis creates novel alleles, thereby increasing genetic and phenotypic diversity. The availability of the faba bean (Vicia faba L.) reference genome and a growing set of additional genomic resources has increased the scientific and practical value of mutant collections. We aimed to genotype and morphologically phenotype a historical faba bean mutant collection developed and characterized by Jan Sjödin (1934-2023) over half a century ago in order to increase its value to researchers. The collection was genotyped using high-throughput single-primer enrichment technology (SPET) assays., Results: We used 11,073 informative single nucleotide polymorphism (SNP) markers spanning the faba bean genome to genotype 52 mutant lines along with the background line, cv. Primus. A range of flower, seed, leaf, and stipule mutations were observed. The analysis of population structure revealed a shallow structure with no major subpopulations. Principal component and cluster analyses revealed, to a minor extent, that the mutants clustered by their phenotype., Conclusions: The mutants' phenotypic variation and shallow structure indicate that the Sjödin faba bean collection has the potential to play a significant role in faba bean breeding and in genetic and functional studies., (© 2024. The Author(s).)

Faba bean is an important pulse. It provides proteins for the human diet and is used in industrial foodstuffs, such as flours. Drought stress severely reduces the yield of faba bean, and this can be efficiently overcome through the identification and application of key genes in response to drought. In this study, PacBio and Illumina RNA sequencing techniques were used to identify the key pathways and candidate genes involved in drought stress response. During seed germination, a total of 17,927 full-length transcripts and 12,760 protein-coding genes were obtained. There were 1676 and 811 differentially expressed genes (DEGs) between the varieties E1 and C105 at 16 h and 64 h under drought stress, respectively. Six and nine KEGG pathways were significantly enriched at 16 h and 64 h under drought stress, which produced 40 and 184 nodes through protein-protein interaction (PPI) analysis, respectively. The DEGs of the PPI nodes were involved in the ABA (abscisic acid) and MAPK (mitogen-activated protein kinase) pathways, N-glycosylation, sulfur metabolism, and sugar metabolism. Furthermore, the ectopic overexpression of a key gene, AAT , encoding aspartate aminotransferase (AAT), in tobacco, enhanced drought tolerance. The activities of AAT and peroxidase (POD), the contents of cysteine and isoleucine, were increased, and the contents of malonaldehyde (MDA) and water loss decreased in the overexpressed plants. This study provides a novel insight into genetic response to drought stress and some candidate genes for drought tolerance genetic improvements in this plant.

Faba bean is an important legume crop. The genetic diversity among faba bean genotypes is very important for the genetic improvement of target traits. A set of 128 fab bean genotypes that are originally from Egypt were used in this study to investigate the genetic diversity and population structure. The 128 genotypes were genotyped using the Single Primer Enrichment Technology (SPET) by which a set of 6759 SNP markers were generated after filtration. The SNP markers were distributed on all chromosomes with a range extending from 822 (Chr. 6) to 1872 (Chr.1). The SNP markers had wide ranges of polymorphic information content (PIC), gene diversity (GD), and minor allele frequency. The analysis of population structure divided the Egyptian faba bean population into five subpopulations. Considerable genetic distance was found among all genotypes, ranging from 0.1 to 0.4. The highly divergent genotype was highlighted in this study and the genetic distance among genotypes ranged from 0.1 and 0.6. Moreover, the structure of linkage disequilibrium was studied, and the analysis revealed a low level of LD in the Egyptian faba bean population. A slow LD decay at the genomic and chromosomal levels was observed. Interestingly, the distribution of haplotype blocks was presented in each chromosome and the number of haplotype block ranged from 65 (Chr. 4) to 156 (Chr. 1). Migration and genetic drift are the main reasons for the low LD in the Egyptian faba bean population. The results of this study shed light on the possibility of the genetic improvement of faba bean crop in Egypt and conducting genetic association analyses to identify candidate genes associated with target traits (e.g. protein content, grain yield, etc.) in this panel., (© 2024. The Author(s).)

Background: Salinity is a major abiotic stress, and the use of saline water in the agricultural sector will incur greater demand under the current and future climate changing scenarios. The objective of this study was to develop a dual-functional nanofertilizer capable of releasing a micronutrient that nourishes plant growth while enhancing salt stress resilience in faba bean (Vicia faba L.)., Results: Moringa oleifera leaf extract was used to synthesize sulfur nanoparticles (SNPs), which were applied as a foliar spray at different concentrations (0, 25, 50, and 100 mg/l) to mitigate the negative effects of salt stress (150 mM NaCl) on faba bean plants. The SNPs were characterized and found to be spherical in shape with an average size of 10.98 ± 2.91 nm. The results showed that salt stress had detrimental effects on the growth and photosynthetic performance (Fv/Fm) of faba bean compared with control, while foliar spraying with SNPs improved these parameters under salinity stress. SNPs application also increased the levels of osmolytes (soluble sugars, amino acids, proline, and glycine betaine) and nonenzymatic antioxidants, while reducing the levels of oxidative stress biomarkers (MDA and H 2 O 2 ). Moreover, SNPs treatment under salinity stress stimulated the activity of antioxidant enzymes (ascorbate peroxidase (APX), and peroxidase (POD), polyphenol oxidase (PPO)) and upregulated the expression of stress-responsive genes: chlorophyll a-b binding protein of LHCII type 1-like (Lhcb1), ribulose bisphosphate carboxylase large chain-like (RbcL), cell wall invertase I (CWINV1), ornithine aminotransferase (OAT), and ethylene-responsive transcription factor 1 (ERF1), with the greatest upregulation observed at 50 mg/l SNPs., Conclusion: Overall, foliar application of sulfur nanofertilizers in agriculture could improve productivity while minimizing the deleterious effects of salt stress on plants. Therefore, this study provides a strong foundation for future research focused on evaluating the replacement of conventional sulfur-containing fertilizers with their nanoforms to reduce the harmful effects of salinity stress and enhance the productivity of faba beans., (© 2024. The Author(s).)

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).)

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.)

Chocolate spot and heat stress devastatingly impact the production of faba bean, particularly under prevailing climatic changes and rising drastic environmental conditions. Hence, the adaptability of faba bean performance is a decisive objective of plant breeders to ensure its sustainable production. The present study aimed to evaluate the agronomic performance and stability of diverse eleven faba bean genotypes for yield characters, chocolate spot, and heat stress in eight different growing environments. The faba bean genotypes were evaluated at two sowing dates in two different locations during two growing seasons. The evaluated eleven faba bean genotypes were sown timely in autumn (25 October) and late sowing in early winter (25 November) in Bilbeis and Elkhatara during 2020 and 2021 growing seasons. The results exhibited substantial differences among the evaluated sowing dates, locations, and faba bean genotypes for all studied characters. The genotypes Sakha-3, Nubaria-3, Nubaria-5, Misr-3, and Wadi-1 were able to produce acceptable yield and quality characters under timely sowing in autumn and late sowing in early winter in all tested environments. Moreover, the genotypes Nubaria-3, Nubaria-4, Nubaria-5, Sakha-4, Giza-3, and Triple White exhibited better resistance to chocolate spot. The assessed faba bean genotypes were evaluated under late sowing to expose the plants to high temperature stress at flowering and throughout the anthesis and seed-filling stages. The genotypes Nubaria-5, Nubaria-3, Nubaria-4, Sakha-3, Sakha-4, Wadi-1, and Misr-3 possessed tolerance to heat stress more than the other genotypes. Different statistical methods were applied to study the stability of assessed genotypes such as joint regression, Additive Main Effect and Multiplicative Interaction (AMMI) analysis, AMMI stability value, Wricke's and Ecovalence values. The estimated stability parameters were consistent in depicting the stability of the assessed faba bean genotypes. The findings revealed that Sakha-1, Misr-3, Nubaria-4, and Nubaria-5 demonstrated stable and desirable performance across all tested environments. The heatmap was employed to classify the assessed faba bean genotypes into different groups based on agronomic performance, chocolate spot resistance and heat stress tolerance. Nubaria-3, Nubaria-4, Nubaria-5, and Misr-3 had the best performance for agronomic performance, chocolate spot resistance, and heat stress tolerance. The obtained results provide evidence of employing promising faba bean genotypes for improving the stability of agronomic performance, chocolate spot resistance, and heat stress tolerance in breeding programs principally under unprecedented climate fluctuations., (© 2024. The Author(s).)

Background: Orobanche is an obligate parasite on faba bean in the Mediterranean region, causes considerable yield losses. Breeding tolerant faba bean genotypes to Orobanche is pivotal to sustain production and ensuring global food security, particularly considering the challenges posed by population growth. In the present study, seven faba bean lines and four testers were used in a line×tester mating design during 2020-2021 and 2021-2022 growing seasons. The eleven parents and their 28 F 1 crosses were evaluated under Orobanche free and naturally infested soils., Results: The results demonstrated considerable variations among the evaluated genotypes, wide diversity among the parental materials, and heterotic effects for all studied agronomic traits under Orobanche-free and infested soils. Orbanche infestation displayed a significant adverse impact on all the studied agronomic traits. The genotypes Line1, Line2, Line3, and Line5 displayed superior performance under Orobanche-infested conditions and recorded the highest values of all studied agronomic traits. Additionally, Line1, Line2, Line3, Line5, and Line7 exhibited desirable significant GCA for most evaluated traits under the two infestation conditions. The obtained crosses displayed significant negative or positive heterosis for studied agronomic characters such as plant height, number of branches per plant, number of pods per plant, number of seeds per plant , and seed weight per plant were observed. Furthermore, specific cross combinations such as Line2×Sakha3, Line3×Nubaria5, Line7 × Nubaria5, Line6×Nubaria1, Line5×Sakha3, Line1×Sakha3, and Line1 × Nubaria5 exhibited superior performance in seed yield and contributing traits under Orobanche-infested conditions. Moreover, these specific crosses showed superior efficacy in reducing dry weight of Orobanche spikes. The results obtained from GGE biplot analysis closely aligned with those from the line×tester procedure, affirming the significance of GGE biplot as a valuable statistical tool for assessing genotype combining ability in line× tester data. Both additive and non-additive gene actions were reported to be predominantly involved in the inheritance of the studied agronomic traits in faba bean., Conclusions: The detected genetic diversity within the evaluated faba bean genotypes and their developed crosses exhibits substantial potential for improving faba bean productivity under Orobanche-infested conditions. The parental genotypes, Line1, Line2, Line3, Line5, and Line7, were identified as effective and promising combiners. Moreover, the developed crosses Line2×Sakha3, Line3×Nubaria5, Line7×Nubaria5, Line6×Nubaria1, Line5×Sakha3, Line1×Sakha3, and Line1×Nubaria5 could be considered valuable candidates for developing high-yielding and tolerant faba bean genotypes to Orobanche., (© 2024. The Author(s).)

The genome of faba bean was first published in 2023. To promote future molecular breeding studies, we improved the quality of the faba genome based on high-density genetic maps and the Illumina and Pacbio RNA-seq datasets. Two high-density genetic maps were used to conduct the scaffold ordering and orientation of faba bean, culminating in an increased length (i.e., 14.28 Mbp) of chromosomes and a decrease in the number of scaffolds by 45. In gene model mining and optimisation, the PacBio and Illumina RNA-seq datasets from 37 samples allowed for the identification and correction 121,606 transcripts, and the data facilitated a prediction of 15,640 alternative splicing events, 2,148 lncRNAs, and 1,752 fusion transcripts, thus allowing for a clearer understanding of the gene structures underlying the faba genome. Moreover, a total of 38,850 new genes including 56,188 transcripts were identified compared with the reference genome. Finally, the genetic data of the reference genome was integrated and a comprehensive and complete faba bean transcriptome sequence of 103,267 transcripts derived from 54,753 uni-genes was formed., (© 2024. The Author(s).)

Key Message: Low concentrations of hydroxyurea, an inhibitor of DNA replication, induced oxidative and replicative stress in root apical meristem (RAM) cells of Vicia faba. Plant cells are constantly exposed to low-level endogenous stress factors that can affect DNA replication and lead to DNA damage. Long-term treatments of Vicia faba root apical meristems (RAMs) with HU leads to the appearance of atypical cells with intranuclear asynchrony. This rare form of abnormality was manifested by a gradual condensation of chromatin, from interphase to mitosis (so-called IM cells). Moreover, HU-treated root cells revealed abnormal chromosome structure, persisting DNA replication, and elevated levels of intracellular hydrogen peroxide (H 2 O 2 ) and superoxide anion (O 2 ∙- ). Immunocytochemical studies have shown an increased number of fluorescent foci of H3 histones acetylated at lysine 56 (H3K56Ac; canonically connected with the DNA replication process). We show that continuous 3-day exposure to low concentrations (0.75 mM) of hydroxyurea (HU; an inhibitor of DNA replication) induces cellular response to reactive oxygen species and to DNA replication stress conditions., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Like other heavy metals, Cr (VI) is a powerful carcinogen and mutagen agent. Its toxic effects on plants are well considered. In order to elucidate its adverse effects, the present work aims to study the mitosis aberrations of Cr (VI) on the Vicia faba root-cells and its molecular docking analysis to understand the genotoxicity mechanisms. In-vivo, Vicia faba plants were exposed to 50 and 100 μM Cr (VI) for 48 h. In-silico, molecular docking and molecular dynamics simulation were used to study the interactions between dichromate and tubulin tyrosine ligase T2R-TTL (PDBID: 5XIW) with reference to Colchicine (microtubule inhibitor). According to our results, Cr (VI) affects growth and cell division and also induces many mitosis aberrations such as chromosome sticking, anaphase/telophase bridges, lagging chromosomes and fragmentation during all phases of mitosis. On the one hand, Cr (VI) reduces mitotic index and promotes micronuclei induction. The in-silico results showed that dichromate establishes very strong bonds at the binding site of the tubulin tyrosine ligase T2R-TTL, with a binding affinity of -5.17 Kcal/Mol and an inhibition constant of 163.59 μM. These interactions are similar to those of colchicine with this protein, so dichromate could be a very potent inhibitor of this protein's activity. TTL plays a fundamental role in the tyrosination/detyrosination of tubulin, which is crucial to the regulation of the microtubule cytoskeleton. Its inhibition leads to the appearance of many morphogenic abnormalities such as mitosis aberrations. In conclusion, our data confirm the highest genotoxicity effects of Cr (VI) on Vicia faba root-cells., 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.)

Differential accumulation of the distinct genome segments is a common feature of viruses with segmented genomes. The reproducible and specific pattern of genome segment accumulation within the host is referred to as the "genome formula". There is speculation and some experimental support for a functional role of the genome formula by modulating gene expression through copy number variations. However, the mechanisms of genome formula regulation have not yet been identified. In this study, we investigated whether the genome formula of the octopartite nanovirus faba bean necrotic stunt virus (FBNSV) is regulated by processes acting at the individual segment vs. viral population levels. We used a leaf infiltration system to show that the two most accumulated genome segments of the FBNSV possess a greater intrinsic accumulation capacity in Vicia faba tissues than the other segments. Nevertheless, processes acting at the individual segment level are insufficient to generate the genome formula, suggesting the involvement of additional mechanisms acting at the supra-segment level. Indeed, the absence of segments with important functions during systemic infection strongly modifies the relative frequency of the others, indicating that the genome formula is a property of the segment group. Together, these results demonstrate that the FBNSV genome formula is shaped by a complex process acting at both the individual segment and the segment group levels., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Bonnamy 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.)

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.)

Importance: Globally, viral diseases impair the growth and vigor of cultivated crops such as grains, leading to a significant reduction in quality, marketability, and competitiveness. As an island nation, Australia has a distinct advantage in using its border to prevent the introduction of damaging viruses, which threaten the continental agricultural sector. However, breeding programs in Australia rely on imported seeds as new sources of genetic diversity. As such, it is critical to remain vigilant in identifying new and emerging viral pathogens, by ensuring the availability of accurate genomic diagnostic tools at the grain biosecurity border. High-throughput sequencing offers game-changing opportunities in biosecurity routine testing. Genomic results are more accurate and informative compared to traditional molecular methods or biological indexing. The present work contributes to strengthening accurate phytosanitary screening, to safeguard the Australian grains industry, and expedite germplasm release to the end users., Competing Interests: The authors declare no conflict of interest.

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).)

Reference genomes facilitate trait improvement by aiding in the elucidation of causal genetic elements. Thanks to the recent release of a reference sequence for the faba bean, breeders and geneticists are poised to accelerate precision breeding and genetic improvement of this important crop., Competing Interests: Declaration of interest The authors declare no conflict of interest., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Cucumber mosaic virus (CMV) is one of the most prevalent plant viruses in the world, and causes severe damage to various crops. CMV has been studied as a model RNA virus to better understand viral replication, gene functions, evolution, virion structure, and pathogenicity. However, CMV infection and movement dynamics remain unexplored due to the lack of a stable recombinant virus tagged with a reporter gene. In this study, we generated a CMV infectious cDNA construct tagged with a variant of the flavin-binding LOV photoreceptor (iLOV). The iLOV gene was stably maintained in the CMV genome after more than four weeks of three serial passages between plants. Using the iLOV-tagged recombinant CMV, we visualized CMV infection and movement dynamics in living plants in a time course manner. We also examined whether CMV infection dynamics is influenced by co-infection with broad bean wilt virus 2 (BBWV2). Our results revealed that no spatial interference occurred between CMV and BBWV2. Specifically, BBWV2 facilitated the cell-to-cell movement of CMV in the upper young leaves. In addition, the BBWV2 accumulation level increased after co-infection with CMV., (© 2023. The Author(s).)

Faba bean is an important legume crop consumed as a vegetable or snack food, and its green cotyledons could present an attractive color for consumers. A mutation in SGR causes stay-green in plants. In this study, vfsgr was identified from a green-cotyledon-mutant faba bean, SNB7, by homologous blast between the SGR of pea and the transcriptome of faba bean. Sequence analysis revealed that a SNP at position 513 of the CDS of VfSGR caused a pre-stop codon, resulting in a shorter protein in the green-cotyledon faba bean SNB7. A dCaps marker was developed according to the SNP that caused the pre-stop, and this marker was completely associated with the color of the cotyledon of faba bean. SNB7 stayed green during dark treatment, while the expression level of VfSGR increased during dark-induced senescence in the yellow-cotyledon faba bean HST. Transient expression of VfSGR in Nicotiana. benthamiana leaves resulted in chlorophyll degradation. These results indicate that vfsgr is the gene responsible for the stay-green of faba bean, and the dCaps marker developed in this study provides a molecular tool for the breeding of green-cotyledon faba beans.

Key Message: We identified marker-trait associations for key faba bean agronomic traits and genomic signatures of selection within a global germplasm collection. Faba bean (Vicia faba L.) is a high-protein grain legume crop with great potential for sustainable protein production. However, little is known about the genetics underlying trait diversity. In this study, we used 21,345 high-quality SNP markers to genetically characterize 2678 faba bean genotypes. We performed genome-wide association studies of key agronomic traits using a seven-parent-MAGIC population and detected 238 significant marker-trait associations linked to 12 traits of agronomic importance. Sixty-five of these were stable across multiple environments. Using a non-redundant diversity panel of 685 accessions from 52 countries, we identified three subpopulations differentiated by geographical origin and 33 genomic regions subjected to strong diversifying selection between subpopulations. We found that SNP markers associated with the differentiation of northern and southern accessions explained a significant proportion of agronomic trait variance in the seven-parent-MAGIC population, suggesting that some of these traits were targets of selection during breeding. Our findings point to genomic regions associated with important agronomic traits and selection, facilitating faba bean genomics-based breeding., (© 2023. The Author(s).)

Background and Aims: Limiting maximum transpiration rate (TR) under high vapour pressure deficit (VPD) works as a water conservation strategy. While some breeding programmes have incorporated this trait into some crops to boost yields in water-limited environments, its underlying physiological mechanisms and genetic regulation remain unknown for faba bean (Vicia faba). Thus, we aimed to identify genetic variation in the TR response to VPD in a population of faba bean recombinant inbred lines (RILs) derived from two parental lines with contrasting water use (Mélodie/2 and ILB 938/2)., Methods: Plants were grown in well-watered soil in a climate-controlled glasshouse with diurnally fluctuating VPD and light conditions. Whole plant transpiration was measured in a gas exchange chamber that tightly regulated VPD around the shoot under constant light, while whole-plant hydraulic conductance and its components (root and stem hydraulic conductance) were calculated from dividing TR by water potential gradients measured with a pressure chamber., Key Results: Although TR of Mélodie/2 increased linearly with VPD, ILB 938/2 limited its TR above 2.0 kPa. Nevertheless, Mélodie/2 had a higher leaf water potential than ILB 938/2 at both low (1.0 kPa) and high (3.2 kPa) VPD. Almost 90 % of the RILs limited their TR at high VPD with a break-point (BP) range of 1.5-3.0 kPa and about 10 % had a linear TR response to VPD. Thirteen genomic regions contributing to minimum and maximum transpiration, and whole-plant and root hydraulic conductance, were identified on chromosomes 1 and 3, while one locus associated with BP transpiration was identified on chromosome 5., Conclusions: This study provides insight into the physiological and genetic control of transpiration in faba bean and opportunities for marker-assisted selection to improve its performance in water-limited environments., (© The Author(s) 2023. Published by Oxford University Press on behalf of the Annals of Botany Company.)

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: Faba bean ( Vicia faba L) is one of the most important legumes in the world. However, there is relatively little genomic information available for this species owing to its large genome. The lack of data impedes the discovery of molecular markers and subsequent genetic research in faba bean. The objective of this study was to analyze the faba bean transcriptome, and to develop simple sequence repeat (SSR) markers to determine the genetic diversity of 226 faba bean varieties derived from different regions in China., Methods: Faba bean varieties with different phenotype were used in transcriptome analysis. The functions of the unigenes were analyzed using various database. SSR markers were developed and the polymorphic markers were selected to conduct genetic diversity analysis., Results: A total of 92.43 Gb of sequencing data was obtained in this study, and 133,487 unigene sequences with a total length of 178,152,541 bp were assembled. A total of 5,200 SSR markers were developed on the basis of RNA-Seq analysis. Then, 200 SSR markers were used to evaluate polymorphisms. In total, 103 (51.5%) SSR markers showed significant and repeatable bands between different faba bean varieties. Clustering analysis revealed that 226 faba bean materials were divided into five groups. Genetic diversity analysis revealed that the relationship between different faba beans in China was related, especially in the same region. These results provided a valuable data resource for annotating genes to different categories and developing SSR markers., Competing Interests: The authors declare there are no competing interests., (©2023 Hou et al.)

Nanoviruses are plant multipartite viruses with a genome composed of six to eight circular single-stranded DNA segments. The distinct genome segments are encapsidated individually in icosahedral particles that measure ≈18 nm in diameter. Recent studies on the model species Faba bean necrotic stunt virus (FBNSV) revealed that complete sets of genomic segments rarely occur in infected plant cells and that the function encoded by a given viral segment can complement the others across neighbouring cells, presumably by translocation of the gene products through unknown molecular processes. This allows the viral genome to replicate, assemble into viral particles and infect anew, even with the distinct genome segments scattered in different cells. Here, we question the form under which the FBNSV genetic material propagates long distance within the vasculature of host plants and, in particular, whether viral particle assembly is required. Using structure-guided mutagenesis based on a 3.2 Å resolution cryogenic-electron-microscopy reconstruction of the FBNSV particles, we demonstrate that specific site-directed mutations preventing capsid formation systematically suppress FBNSV long-distance movement, and thus systemic infection of host plants, despite positive detection of the mutated coat protein when the corresponding segment is agroinfiltrated into plant leaves. These results strongly suggest that the viral genome does not propagate within the plant vascular system under the form of uncoated DNA molecules or DNA:coat-protein complexes, but rather moves long distance as assembled viral particles., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Trapani 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.)

Purpose: Three experiments were conducted to assess the effect of different doses of gamma radiation on various seedling traits; determine the optimum doses of gamma radiation for different faba bean genotypes; find out the variation in optimum doses with respect to the different times of sowings after irradiation and methods of irradiation., Materials and Methods: Five faba bean genotypes viz ., L-2013-060, L-2013-092, Anandnagar Local, Gazipur Local and Bangla Gangachar were used in these experiments. In Experiment I, seeds of five experimental genotypes were exposed to different doses (100 Gy 200 Gy, 300 Gy, 400 Gy, 500 Gy, 600 Gy, 700 Gy and 800 Gy) of gamma radiation and were sown immediately after irradiation. In Experiment II, seeds of Bangla Gangachar and L-2013-060 were exposed to varying doses (100-800 Gy) of gamma radiation and were sown at seven sowings starting from 0 h to 24 h at 4-h intervals after irradiation. In Experiment III, L-2013-092 genotypes was exposed to different doses (100 -800 Gy) of gamma radiation with two different methods of irradiation., Results: In Experiment I, the lethal dose 50 (LD 50 ) values have arrived at 140 Gy, 669 Gy, 575 Gy, 386 Gy and 158 Gy for L-2013-060, L-2013-092, Anandnagar Local, Gazipur Local and Bangla Gangachar, respectively. The growth reduction 50 (GR 50 ) doses for different seedling traits ranged from 130 Gy to 320 Gy for L-2013-060, 250 Gy to 480 Gy for L-2013-092, 130 Gy to 370 Gy for Anandnagar Local, 200 Gy to 350 Gy for Gazipur Local and 250 Gy to 400 Gy for Bangla Gangachar. In Experiment II, the values for LD 50 of the genotypes Bangla Gangachar and L-2013-060 were significantly singular for different time intervals of sowing. The values of GR 50 for most of the seedling traits were found to increase with the delay in sowing after irradiation from 4 to 24 h when compared with the immediately sown seed lots. In Experiment III, LD 50 for L-2013-092 was 337 Gy with Method 1 and 669 Gy with Method 2. In Method 1, most of the growth parameters attained GR 50 doses lower than Method 2. The first method was found to increase the radiosensitivity of L-2013-092., Conclusion: Every experimental genotype used in these three experiments showed dose-dependent retardation of different seedling traits. These optimized doses may be employed to establish mutant populations for exploiting the novel traits of faba bean. The time of sowing after irradiation and method of irradiation was found to be essential for confirming optimum doses.

Crisp grass carp (CGC), the most representative improved varieties of grass carp (GC), features higher muscle hardness after feeding faba bean ( Vicia faba L.) for 90-120 days. DNA methylation, a most widely studied epigenetic modification, plays an essential role in muscle development. Previous studies have identified numerous differentially expressed genes (DEGs) between CGC and GC. However, it remains unknown if the expression levels of these DEGs are influenced by DNA methylation. In the present study, we performed a comprehensive analysis of DNA methylation profiles between CGC and GC, and identified important candidate genes related to muscle development coupled with the transcriptome sequencing data. A total of 9,318 differentially methylated genes (DMGs) corresponding to 155,760 differentially methylated regions (DMRs) were identified between the two groups under the CG context in promoter regions. Combined with the transcriptome sequencing data, 14 key genes related to muscle development were identified, eight of which (gsk3b, wnt8a, wnt11, axin2, stat1, stat2, jak2, hsp90) were involved in muscle fiber hyperplasia, six of which (tgf-β1, col1a1, col1a2, col1a3, col4a1, col18a1) were associated with collagen synthesis in crisp grass carp. The difference of methylation levels in the key genes might lead to the expression difference, further resulting in the increase of muscle hardness in crisp grass carp. Overall, this study can help further understand how faba bean modulates muscle development by the epigenetic modifications, providing novel insights into the texture quality improvement in other aquaculture fish species by nutritional programming., Competing Interests: The authors declare that they have no competing interests., (© 2022 Li et al.)

Faba bean (Vicia faba) is a grain legume crop widely cultivated in temperate areas for food and feed. Its productivity can be constrained by numerous diseases and pests that can be managed by a number of strategies, complemented with the deployment of resistant cultivars in an integrated manner. Few sources of resistance are available to some of them, although their phenotypic expression is usually insufficiently described, and their genetic basis is largely unknown. A few DNA markers have been developed for resistance to rust, ascochyta blight, and broomrape, but not yet for other diseases or pests. Still, germplasm screenings are allowing the identification of resistances that are being accumulated by classical breeding, succeeding in the development of cultivars with moderate levels of resistance. The adoption of novel phenotyping approaches and the unprecedented development of genomic resources along with speed breeding tools are speeding up resistance characterization and effective use in faba bean breeding., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Faba bean ( Vicia faba L.), a drought-sensitive crop, is drastically affected by drought stresses compromising its growth and yield. However, wild relatives of faba bean are considered a reservoir of potential genetic resources for tolerance against abiotic stresses. This study was conducted to characterize wild relatives of faba bean for identification of a specific tolerance system required for its improvement against drought stress. The study focused on physiological, biochemical, and anatomical responses of wild Vicia species under drought stress conditions. The experiment was carried out under various levels of drought stress imposed through different field capacities (FC) which included 80% FC ie (well-watered condition), 55% FC (moderate stress), and 30% FC (severe stress). When compared to plants grown in a control environment, drought stress significantly reduced the studied physiological attributes including soluble sugars (21.3% and 15.8%), protein contents (14.7 and 14.6%), and chlorophyll (8.4 and 28.6%) under moderate (55% FC) and severe drought stress (30% FC), respectively. However, proline content increased by 20.5% and 27.6%, peroxidase activity by 48.5% and 57.1%, and superoxide dismutase activity by 72.6% and 64.8% under moderate and severe stress, respectively. The studied anatomical attributes were also affected under drought stress treatments, including diameter of stem xylem vessels (9.1% and 13.7%), leaf lower epidermal thickness (8.05% and 13.34%), and leaf phloem width (5.3% and 10.1%) under moderate and severe stress, respectively. Wild Vicia spp. showed better tolerance to water-deficit conditions as compared to cultivated Vicia L. The observed potential diversity for drought tolerance in wild Vicia spp. may assist in improvement of faba bean and may also help in understanding the mechanisms of adaptations in drought-prone environments.

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.)

Recent morphological and molecular studies confirmed Physoderma viciae , and not Olpidium viciae , to be the causative agent of the devastating Faba Bean Gall (FBG) disease on faba bean ( Vicia faba ) in Ethiopia and also highlighted its ability to cross-infect with other host genera such as Pisum and Trifolium . In this study, the first pair of specific primer 'Physo 1' and primer pair 'Physo D' are reported from molecular sequences of this pathogen from the conserved LSU (S28) gene. Whereas 'Physo 1' readily detects P. viciae , 'Physo D', clearly separates its identity from the common and confounding presence of Didymella/Phoma spp. The study also reports the presence of the Ascochyta blight pathogen complex, symptomless but almost universal on field pea ( Pisum sativum ), within faba bean infested by P. viciae. We emphasize historical evidence confirming such unique association in other legumes, such as the subterranean clover ( Trifolium subterraneum ). This new finding has significant implications for rotations involving different legume crop and/or forage legume genera and possibly provides the first explanation for the widespread occurrence of the field pea Ascochyta blight pathogen complex even in the absence of field pea cropping for many years.

The mechanisms by which plants respond to alkali salt stress are still obscure, and the relevance of alkaline pH under combined alkali salt stress. Early stress responses can indicate mechanisms leading to damage and plant resistance. The apoplast contains essential determinants for plant growth, specifically early apoplastic pH fluctuations are induced by many stressors and hypothesized to be involved in stress signalling. Hence, this study aims to identify fast responses specific to alkaline pH and alkali salt stress by exposing the root of hydroponically grown Vicia faba L. plants to 150 min of either 50 mM NaHCO 3 (pH 9) treatment or alkaline pH 9 alone. Apoplastic pH was monitored in real-time by ratiometric fluorescence microscopy simultaneously with SWIR transmission-based measurements of leaf water content (LWC). Moreover, we examined the effect of these stresses on apoplastic, symplastic and xylem ion and metabolite composition together with transcriptions of certain stress-responsive genes. Physiological and transcriptional changes were observed in response to NaHCO 3 but not to alkaline pH alone. NaHCO 3 elicited a transient reduction in LWC, followed by a transient alkalinization of the apoplast and stomatal closure. Simultaneously, organic acids and sugars accumulated. Fast upregulation of stress-responsive genes showed the significance of gene regulation for early plant adaptation to alkali salt stress., (Copyright © 2022 Elsevier B.V. All rights reserved.)

In Ethiopia, faba bean ( Vicia faba L.) varieties are important and widely used as a source of food and cash source to large number of subsistence farmers. However, their production and productivity is below the world's average partly because of lack of their sustainable performance in the current scenario of climate change. Therefore, the present study was designed to test the effects of seed storage time and to evaluate the performance of 31 faba bean varieties collected from Holeta, Kulumsa and Sinana agricultural research systems, Ethiopia. The study involved germination test and field experiment that was laid out using randomized complete block design (RCBD) and conducted at multiple test locations. Data were collected on qualitative and quantitative traits and analyzed using SAS version 9.0, and MINITAB® Release 19. Accordingly, most of the varieties showed a promising germination rate regardless of their storage duration suggesting their sustainable performance under suitable storage conditions. Most of the qualitative and quantitative traits showed a wide range of variations revealing their stable performance and better chance for further improvement. Analysis of variance also revealed a highly significant ( p < 0.001) variation for several of the traits suggesting maintenance of the original diversity that could be important in further selection breeding. Likewise, high genetic advance coupled with high heritability and genotypic coefficient of variation together with wide range of variations in both PCV and GCV observed in several of the quantitative traits suggest their sustainable performance and significance in further effective selection. Moreover, a promising high yielding varieties such as Dida1, Welki, Hachalu, Ashebeka and Obse have been identified for further use. Clustering grouped the varieties into three clusters implying significant amount of genetic variability among them. Overall, the results generated could be used as a baseline information for improving faba bean production and productivity. However, to exploit more and determine the actual performance of the varieties more markers such as molecular markers (DNA based) are recommended., Competing Interests: The authors declare that they have no conflicts of interest., (Copyright © 2022 Fekadu Gadissa et al.)

Autofertility describes the ability of faba bean flowers to self-fertilize thereby ensuring the productivity of this crop in the absence of pollinators or mechanical disturbance. In the legume crop faba bean (Vicia faba L.), lack of autofertility in a context of insufficient pollination can lead to a severe decrease in grain yield. Here we performed the first QTL analysis aimed at identifying the genomic regions controlling autofertility in this crop. We combined pod and seed setting scores from a recombinant inbred population (RIL) segregating for autofertility in different environments and years with measurements of morphological floral traits and pollen production and viability. This approach revealed 19 QTLs co-localizing in six genomic regions. Extensive co-localization was evident for various floral features whose QTLs clustered in chrs. I, II and V, while other QTLs in chrs. III, IV and VI revealed co-localization of flower characteristics and pod and seed set data. The percentage of phenotypic variation explained by the QTLs ranged from 8.9 for style length to 25.7 for stigma angle. In the three QTLs explaining the highest phenotypic variation (R 2  > 20), the marker alleles derived from the autofertile line Vf27. We further inspected positional candidates identified by these QTLs which represent a valuable resource for further validation. Our results advance the understanding of autofertility in faba bean and will aid the identification of responsible genes for genomic-assisted breeding in this crop., (© 2022. The Author(s).)

In Tunisia, drought stress is a major environmental factor limiting crop production and causing relatively low and unstable faba bean yields. In the present study, we explored the putative role of spermidine (0.5, 1, 1.5 and 2mM) in ameliorating the effects of drought stress induced by polyethylene glycol (PEG-6000, -0.58MPa) in faba bean seedlings. Drought stress reduced photosynthetic performance, chlorophyll and relative water content in leaves of faba bean variety Badii. Moreover, drought increased proline, electrolyte leakage and malondialdehyde content by inducing reactive oxygen species (hydrogen peroxide) generation in leaves. However, applying spermidine increased the activities of catalase, superoxide dismutase, ascorbate peroxidase and guaiacol peroxidase. The results show that the application of spermidine especially at a rate of 1.5mM effectively reduces oxidative damage and alleviates negative effects caused by drought stress. In addition, exogenous spermidine increased the expression of polyamine biosynthetic enzymes' genes (VfADC , VfSAMDC and VfSPDS ), and reduced the expression of VfSPMS suggesting that exogenous spermidine can regulate polyamines' metabolic status under drought challenge, and consequently may enhance drought stress tolerance in faba bean. Real-time quantitative polymerase chain reaction analysis revealed that some drought responsive genes (VfNAC , VfHSP , VfNCED , VfLEA , VfCAT , VfAPX , VfRD22 , VfMYB , VfDHN , VfERF , VfSOD and VfWRKY ) from various metabolic pathways were differentially expressed under drought stress. Overall, these genes were more abundantly transcribed in the spermidine-treated plants compared to untreated suggesting an important role of spermidine in modulating faba bean drought stress response and tolerance.

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.)

Weeds represent one of the major constraints for faba bean crop. The identification of molecular markers associated with key genes imparting tolerance to herbicides can facilitate and fasten the efficient and effective development of herbicide tolerant cultivars. We phenotyped 140 faba bean genotypes in three open field experiments at two locations in Lebanon and Morocco against three herbicide treatments (T1 metribuzin 250 g ai/ha; T2 imazethapyr 75 g ai/ha; T3 untreated) and one in greenhouse where T1 and T3 were applied. The same set was genotyped using genotyping by sequencing (GBS) which yield 10,794 high quality single nucleotide polymorphisms (SNPs). ADMIXTURE software was used to infer the population structure which revealed two ancestral subpopulations. To identify SNPs associated with phenological and yield related traits under herbicide treatments, Single-trait (ST) and Multi-trait (MT) Genome Wide Association Studies (GWAS) were fitted using GEMMA software, showing 10 and 14 highly significant associations, respectively. Genomic sequences containing herbicide tolerance associated SNPs were aligned against the NCBI database using BLASTX tool using default parameters to annotate candidate genes underlying the causal variants. SNPs from acidic endochitinase, LRR receptor-like serine/threonine-protein kinase RCH1, probable serine/threonine-protein kinase NAK, malate dehydrogenase, photosystem I core protein PsaA and MYB-related protein P-like were significantly associated with herbicide tolerance traits., (© 2022. The Author(s).)

&lt;b&gt;Background and Objective:&lt;/b&gt; It is known that any genetic improvement depends on the presence of many genetic variations so that the plant breeder can choose the desired traits such as the trait of the crop or resistance to some diseases. Different or it can be used for selection, whether for yield, early maturity or other characteristics. This study aimed to use gamma irradiation and the chemical mutagen Dimethyl sulfoxide for the induction of genetic variation in two types of beans. &lt;b&gt;Materials and Methods:&lt;/b&gt; The Giza 429 and Misr 1 genotypes were treated with three different concentrations of dimethyl sulfur oxide (1000, 2000 and 3000 ppm) and gamma radiation doses (10, 15 and 20 kr) to perform mutagenesis treatments. &lt;b&gt;Results:&lt;/b&gt; In the first and second seasons of the study, all the mutations resulting from the radiological and chemical mutagenic treatments of the two Egyptian bean cultivars, Giza 429 and Misr 1, were highly significant for all treatments in the following traits: Number of branches/plant, plant height, number of pods/plant with the weight of 100 seeds (g) and seed yield/plant (g). &lt;b&gt;Conclusion:&lt;/b&gt; The benefit of this research was obtained from plants that outperformed the two cultivars Giza 429 and Misr 1 and are considered among the promising plants that can be used in mixed breeding programs or direct selection for high productivity.

Water deficit has devastating impacts on legume production, particularly with the current abrupt climate changes in arid environments. The application of plant growth-promoting rhizobacteria (PGPR) is an effective approach for producing natural nitrogen and attenuating the detrimental effects of drought stress. This study investigated the influence of inoculation with the PGPR Rhizobium leguminosarum biovar viciae (USDA 2435) and Pseudomonas putida (RA MTCC5279) solely or in combination on the physio-biochemical and agronomic traits of five diverse Vicia faba cultivars under well-watered (100% crop evapotranspiration [ETc]), moderate drought (75% ETc), and severe drought (50% ETc) conditions in newly reclaimed poor-fertility sandy soil. Drought stress substantially reduced the expression of photosynthetic pigments and water relation parameters. In contrast, antioxidant enzyme activities and osmoprotectants were considerably increased in plants under drought stress compared with those in well-watered plants. These adverse effects of drought stress reduced crop water productivity (CWP) and seed yield-related traits. However, the application of PGPR, particularly a consortium of both strains, improved these parameters and increased seed yield and CWP. The evaluated cultivars displayed varied tolerance to drought stress: Giza-843 and Giza-716 had the highest tolerance under well-watered and moderate drought conditions, whereas Giza-843 and Sakha-4 were more tolerant under severe drought conditions. Thus, co-inoculation of drought-tolerant cultivars with R. leguminosarum and P. putida enhanced their tolerance and increased their yield and CWP under water-deficit stress conditions. This study showed for the first time that the combined use of R. leguminosarum and P. putida is a promising and ecofriendly strategy for increasing drought tolerance in legume crops., (© 2021. The Author(s).)

Experiments on Vicia faba root meristem cells exposed to 150 µM cadmium chloride (CdCl 2 ) were undertaken to analyse epigenetic changes, mainly with respect to DNA replication stress. Histone modifications examined by means of immunofluorescence labeling included: (1) acetylation of histone H3 on lysine 56 (H3K56Ac), involved in transcription, S phase, and response to DNA damage during DNA biosynthesis; (2) dimethylation of histone H3 on lysine 79 (H3K79Me2), correlated with the replication initiation; (3) phosphorylation of histone H3 on threonine 45 (H3T45Ph), engaged in DNA synthesis and apoptosis. Moreover, immunostaining using specific antibodies against 5-MetC-modified DNA was used to determine the level of DNA methylation. A significant decrease in the level of H3K79Me2, noted in all phases of the CdCl 2 -treated interphase cell nuclei, was found to correspond with: (1) an increase in the mean number of intranuclear foci of H3K56Ac histones (observed mainly in S-phase), (2) a plethora of nuclear and nucleolar labeling patterns (combined with a general decrease in H3T45Ph), and (3) a decrease in DNA methylation. All these changes correlate well with a general viewpoint that DNA modifications and post-translational histone modifications play an important role in gene expression and plant development under cadmium-induced stress conditions.

Background: Ascochyta blight caused by Ascochyta fabae Speg. and broomrape (Orobanche crenata) are among the economically most significant pathogens of faba bean. Several QTLs conferring resistance against the two pathogens have been identified and validated in different genetic backgrounds. The aim of this study was to saturate the most stable QTLs for ascochyta and broomrape resistance in two Recombinant Inbred Line (RIL) populations, 29H x Vf136 and Vf6 x Vf136, to identify candidate genes conferring resistance against these two pathogens., Results: We exploited the synteny between faba bean and the model species Medicago truncatula by selecting a set of 219 genes encoding putative WRKY transcription factors and defense related proteins falling within the target QTL intervals, for genotyping and marker saturation in the two RIL populations. Seventy and 50 of the candidate genes could be mapped in 29H x Vf136 and Vf6 x Vf136, respectively. Besides the strong reduction of the QTL intervals, the mapping process allowed replacing previous dominant and pedigree-specific RAPD flanking markers with robust and transferrable SNP markers, revealing promising candidates for resistance against the two pathogens., Conclusions: Although further efforts in association mapping and expression studies will be required to corroborate the candidate genes for resistance, the fine-mapping approach proposed here increases the genetic resolution of relevant QTL regions and paves the way for an efficient deployment of useful alleles for faba bean ascochyta and broomrape resistance through marker-assisted breeding., (© 2021. The Author(s).)

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).)

Key Message: Large-scale faba bean transcriptome data are available, and the first genotyping platform based on liquid-phase probe targeted capture technology was developed for genetic and molecular breeding studies. Faba bean (Vicia faba L., 2n = 12) is an important food legume crop that is widely grown for multiple uses worldwide. However, no reference genome is currently available due to its very large genome size (approximately 13 Gb) and limited single nucleotide polymorphism (SNP) markers as well as highly efficient genotyping tools have been reported for faba bean. In this study, 16.7 billion clean reads were obtained from transcriptome libraries of flowers and leaves of 102 global faba bean accessions. A total of 243,120 unigenes were de novo assembled and functionally annotated. Moreover, a total of 1,579,411 SNPs were identified and further filtered according to a selection pipeline to develop a high-throughput, flexible, low-cost Faba&#95;bean&#95;130K targeted next-generation sequencing (TNGS) genotyping platform. A set of 69 Chinese faba bean accessions were genotyped with the TNGS genotyping platform, and the average mapping rate of captured reads to reference transcripts was 93.14%, of which 53.23% were located in the targeted regions. The TNGS genotyping results were validated by Sanger sequencing and the average consistency rate reached 93.6%. Comprehensive population genetic analysis was performed on the 69 Chinese faba bean accessions and identified four genetic subgroups correlated with the geographic distribution. This study provides valuable genomic resources and a reliable genotyping tool that could be implemented in genetic and molecular breeding studies to accelerate new cultivar development and improvement in faba bean., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

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).)

To our knowledge, there was no study on interactive relationship between CHO molecular structure spectral profiles of newly developed cool-season adapted faba bean and nutritional characteristics in ruminants. The aim of this study was to evaluate the impact of genotypes and tannin levels on the physicochemical and nutritional characteristics of faba bean as an alternative protein and energy source for ruminants and its relation to CHO molecular structure spectral profiles using vibrational molecular spectroscopy (Ft/IR-ATR). Eight genotypes with two tannin levels (low and normal) grown at three different locations in Saskatchewan (CDC crop research fields) were analysed. Chemical analyses were performed using AOAC standards and energy was evaluated using NRC standards. Rumen degradation kinetics were determined using in situ dairy trial. Intestinal digestion was analysed by a modified three-step in vitro technique with 12 h pre-rumen incubation in dairy cows. Molecular spectral study was performed using Ft/IR-ATR, and the molecular structural features were analysed at ca. 4000-800 cm -1 using the Spectra ManagerII. CHO-related functional groups were analysed with OMNIC 7.3. The results showed low-tannin faba bean (LT) presented a highly significant difference (p < 0.05) on rumen bypass starch (BSt) compared to normal tannin faba bean (NT). On the other hand, NT had more total tract digested starch (TDST) compared to LT. No significant differences (p > 0.10) were observed for total digestible nutrients (TDN 1x ), metabolizable protein (MP), feed milk value (FMV) or rumen undegraded crude protein (RUP). Results showed differences in physicochemical characteristics among faba bean varieties; however, the predicted production performance was not different within faba bean genotypes. These outcomes suggest that faba bean can be used as nutritive ingredient for dairy cattle without a significant genotype or tannin level effect on metabolic characteristics. Results from vibrational spectroscopic study showed a higher ATR-Ft/IR absorbance (p < 0.05) in NT on inherent structural CHO (STCHO), total CHO (TCHO) area and peaks (H&#95;1015, H&#95;1076, H&#95;1145), and cellulosic compounds (CEC) to total CHO (TCHO) ratio. Significant correlations could be found between CHO spectral profiles and metabolic characteristics of faba bean, which indicates that structural spectral features of faba bean could be used to predict metabolic characteristics in ruminants., (© 2021 Wiley-VCH GmbH.)

The goal of this research was to develop a new genetic database of simple sequence repetition (SSR) primers for faba and classify them according to their target genes and respective biological processes. Approximately 75,605 and 148,196 previously published genomic and transcriptomic faba sequences, respectively, have been used to detect possible SSRs. The numbers of identified SSRs from each dataset were 25,502 and 12,319, respectively. The distribution of different repeat classes indicated that trinucleotides represent the largest number of repeat counts, followed by dinucleotides. The extracted genic SSR sequences were used to design 1091 polymerase chain reaction (PCR) primers, of which only 238 (21.8%) primers target genomic sequences and the other 853 PCR primers targeted transcriptomic sequences. The annotation of gene-targeted SSRs showed that approximately 897 genes were targeted by our SSR primers. Approximately 1890 gene ontology (GO) identification codes have been obtained. The GO keywords were distributed among distinct molecular cell features. The highest redundancies involved 554 technical words, 196 domains, and 160 molecular feature phrases. These GO codes belonged to the general level of GO and included molecular function, cellular component, and biological process (544, 670, and 676 GOs, respectively). Twenty-seven SSR PCR primers were synthesized to 12 Egyptian faba bean genotypes. Approximately 11 SSR provided one to two PCR bands, whereas other SSRs provided only one sharp band with polymorphic band size. There were 13 polymorphic primers. The polymorphism information content was 0.3, which implied moderate informativeness., (© 2021. Institute of Plant Genetics, Polish Academy of Sciences, Poznan.)