Reverse genetic approaches have been widely applied to study gene function in crop species; however, these techniques, including gel-based TILLING, present low efficiency to characterize genes in soybeans due to genome complexity, gene duplication, and the presence of multiple gene family members that share high homology in their DNA sequence. Chemical mutagenesis emerges as a genetically modified-free strategy to produce large-scale soybean mutants for economically important traits improvement. The current study uses an optimized high-throughput TILLING by target capture sequencing technology, or TILLING-by-Sequencing⁺ (TbyS⁺), coupled with universal bioinformatic tools to identify population-wide mutations in soybeans. Four ethyl methanesulfonate mutagenized populations (4032 mutant families) have been screened for the presence of induced mutations in targeted genes. The mutation types and effects have been characterized for a total of 138 soybean genes involved in soybean seed composition, disease resistance, and many other quality traits. To test the efficiency of TbyS⁺ in complex genomes, we used soybeans as a model with a focus on three desaturase gene families, GmSACPD, GmFAD2, and GmFAD3, that are involved in the soybean fatty acid biosynthesis pathway. We successfully isolated mutants from all the six gene family members. Unsurprisingly, most of the characterized mutants showed significant changes either in their stearic, oleic, or linolenic acids. By using TbyS⁺, we discovered novel sources of soybean oil traits, including high saturated and monosaturated fatty acids in addition to low polyunsaturated fatty acid contents. This technology provides an unprecedented platform for highly effective screening of polyploid mutant populations and functional gene analysis. The obtained soybean mutants from this study can be used in subsequent soybean breeding programs for improved oil composition traits.

Sesame (Sesamum indicum L.) is an ancient oilseed, aromatic, and medicinal crop widely used for its high-quality oil and seeds. The available genetic diversity in Morocco is too limited; thus, a mutagenesis breeding program was adopted. This study was carried out to evaluate the novel variability induced and observed in 11 M₄ mutant lines and to estimate some valuable genetic parameters. The experiment was conducted in two different environments using a randomized complete block design with three replications. Phenological, morphological, and agronomic traits were recorded. To estimate the effect of genotype, environment, and their interaction, ANOVA and planned contrast analyses were performed. To examine relatedness among genotypes, cluster analysis was performed. Significant differences among mutants and between parent cultivars and their respective mutant lines were observed. Genetic parameters such as genotypic (GCV) and phenotypic (PCV) coefficients of variation, broad-sense heritability (H² b.s), genetic advance (GA), and genetic advance over the mean (GAM) were high in most traits. Highly productive mutants, ‘US2-1’ and ‘US1-2’, were observed, exhibiting the highest number of capsules per plant and seed yield ever reported. Additionally, there are other promising mutants with early flowering, early maturity, and a reduced height of the first capsule. This suggests that mutagenesis can be successfully applied to develop high-yielding sesame varieties along with other improved phenological and agromorphological traits. All these mutant lines can be used as promising germplasm to develop competitive sesame cultivars to meet the increasing demand for sesame oil and seeds in the actual context of climate change.

Mutagenesis breeding via ethyl methanesulfonate (EMS) has been successfully used in faba bean to improve some economically important traits. However, there is a knowledge gap of the factors/mechanisms related to its sensitivity/tolerance to EMS treatment toxicity. It was hypothesized that the seed size could influence the response of the diverse botanical varieties of faba bean. Consequently, we conducted a comprehensive assessment of the sensitivity of six faba bean varieties: three major varieties (Aguadulce superlonga, Reina mora, and Yasmine) and three minor varieties (Zina, Alfia 05, and Alfia 17), to three increasing concentrations (0.05%, 0.5%, and 1%, along with a control 0%) of EMS. Analyses included various germination parameters (germination percentage [GP], germination energy at seven and 14 days [GE7 and GE14], germination rate index [GRI], and vigor index [VI]) across different EMS concentrations. To further explore mechanisms involved in sensitivity to EMS, we measured coat thickness and assessed antioxidant activity. Our findings revealed that the seed size variation did not significantly affect EMS sensitivity. Different varieties showed significant responses to increasing EMS concentrations ( p ≤ 0.05) for most parameters, except for root length. This challenges the assumption that the seed size influences EMS sensitivity in faba bean. Coat thickness was uniform, suggesting similar EMS absorption. DPPH assay revealed significant antioxidant activity differences between nontreated and EMS‐treated groups. Antioxidant activity correlated significantly with germination parameters under EMS treatment. The study indicates factors beyond seed size contribute to EMS responses. Examining antioxidant systems may explain plants’ ability to counteract EMS‐induced oxidative stress.

Faba bean (Vicia faba L.) is a leguminous crop renowned for its millennia-long cultivation driven by its highly nutritious seeds. Rich in minerals, bioactive carbohydrates, fiber and protein with a complete amino acids profile, faba beans serve as vital components in both human and animal diets. In Morocco, faba bean is crucial for food security, sustainability and rural economy. After stable cultivation in the 1960s-70s and subsequent expansion, recent years have seen a decline in cultivated areas due to market volatility, climate challenges, traditional farming methods, pests and limited research. Addressing these multifaceted challenges requires a comprehensive approach. Key strategies include the adoption of alternative agricultural practices such as no-till farming to enhance soil quality and reduce labor costs. Implementing effective weed management strategies and increasing mechanization are crucial for reducing labor requirements and improving overall productivity. Additionally, breeding programs focused on developing drought-tolerant and high-yielding faba bean varieties are essential. Preparing solutions for emerging pests like aphids and pod borers, which thrive in the shifting hotter and drier climate, is critical. Adopting regional/agroecological varieties and developing open-pollinated and synthetic varieties for the national catalog are important considerations. To combat crenate orobanche, extensive screening of accessions and developing herbicide-resistant varieties should be prioritized. The integration of molecular tools, such as marker-assisted selection and genomic prediction, is crucial for accelerating the breeding process and enhancing precision in developing resilient varieties. Finally, emphasizing scientific research to optimize agronomic practices and reinforce crop resilience stands as a critical priority. This review evaluates the current status of the faba bean sector in Morocco, identifying the obstacles faced by farmers amidst economic, environmental and market fluctuations. It proposes essential strategies aimed at advancing faba bean production in Morocco, serving as a fundamental resource and reference for stakeholders including farmers, researchers, seed producers and policymakers.

Background: Faba bean is a crop of agro-ecological and socio-economic importance in Morocco. Thus, in a changing climatic context, it is important to develop varieties that are resilient to several abiotic constraints, such as synthetics. However, the reproductive system must be understood. Methods: We aimed to estimate faba beans outcrossing rate under natural Moroccan conditions, using two varieties, ‘Reina Mora’ and ‘Zina’ and two phenotypic markers, seed coat color and its size and shape, in two contrasted locations, Saïs and Gharb. Varieties were sown in six alternate rows with six seeds per row and three replicates per location. The experiment consisted in counting the number of purple seeds within ‘Zina’s beige offspring. Besides, seeds collected from ‘Reina Mora’ were counted and processed for differences in size and shape. In total, 859 seeds were measured, including their length, width, thickness, geometric diameter, area and sphericity, enabling to distinguish hybrids from parental seeds and self-seeds. Result: Results showed an average inter-crossing of 3.7% in Saïs and 7.22% in Gharb. A more reliable open-pollination could be achieved by introducing bees and bumblebees colonies as well as using varieties from same geographic gene pools.

Drought is a critical constraint for legume production in semi-arid regions, yet breeding for drought tolerance in faba bean through induced mutagenesis remains largely unexplored. To our knowledge, this is the first EMS-derived mutant population in faba bean specifically developed for drought tolerance, comprising 45 M₂/M₃ lines derived from small-seeded cv. Zina and large-seeded cv. Aguadulce Superlonga), evaluated under two irrigation regimes—100% field capacity (well-watered control) and 40% field capacity (severe stress)—over two consecutive growing seasons in a randomized complete block design with three replications. Drought stress caused severe yield losses, reducing mean seed number per plant by 42.2% and mean seed weight per plant by 47.1%. Analysis of variance revealed highly significant effects of genotype, irrigation, and generation/year on both yield components. The non-significant genotype × irrigation interaction indicated similar proportional drought response across genotypes, while the non-significant three-way interaction suggested relatively consistent genotype rankings across generations/growing seasons. Among the ten drought tolerance indices evaluated, seed-number-based mean productivity (MPn) and stress tolerance index (STIn) were the most discriminating, whereas weight-based indices failed to differentiate genotypes due to the inherent seed-size contrast between botanical backgrounds. Dunnett’s comparisons identified genotype 23 (Zina-derived) as the top performer, significantly exceeding its parent for both MPn and STIn; genotypes 22, 24, 12, 3, and 15 similarly outperformed controls. Cluster analysis broadly distinguished three groups: a tolerant cluster dominated by Zina-derived lines, a moderately tolerant cluster (Zina wild-type), and a sensitive cluster of Aguadulce Superlonga-derived lines. These findings suggest that EMS mutagenesis generated potentially heritable and exploitable variation for drought tolerance, with selected lines representing promising candidates for further multi-environment validation.