Ascochyta pinodes, the leading cause of Ascochyta blight in pea, poses a significant threat to crop yield and quality; nevertheless, limited investigations have utilized metabolomics to study the Pisum sativum-A. pinodes pathosystem. This study presents the first in planta metabolomics-based study focused on the detection of phytotoxic metabolites produced by A. pinodes. Among the fungal metabolites screened, only pinolidoxin was detected in infected tissues, peaking at 48 h post-inoculation. This temporal association suggests a possible effector-like role for pinolidoxin in promoting early infection through interference with host signaling pathways. Furthermore, non-targeted metabolomics-based analysis of A. pinodes infection in susceptible and resistant pea cultivars, Messire and Radley, was also carried out, revealing cultivar-specific metabolic responses in the susceptible cultivar. Messire showed strong activation of phenylpropanoid metabolism, especially flavonoids and hydroxycinnamic acids, while the resistant cv. Radley upregulated amino acids and derivatives, suggesting differences in the defense strategy based on adjustments to primary metabolism, which could be related to Radley resistance. This work highlights the complexity of host-pathogen interactions. It underscores the importance of multi-omics approaches in identifying molecular targets for gaining deeper systems-level insight into pea defense strategies, thereby supporting the development of improved disease-resistant legume varieties.

Aphanomyces root rot is a major threat to legume production worldwide, mainly in pea and lentil, crops on which extensive research programs are targeting the management of the disease. However, other legumes such as common vetch, although known to be severely affected by the disease, remain largely unexplored. This study aimed to identify sources of resistance within V. sativa subsp. sativa accessions. A total of 211 genetically diverse accessions were screened under controlled conditions following inoculation with isolate RB84. Disease progression was monitored through periodic foliar assessments and final root symptom evaluation. To assess resistance stability, a subset of 13 accessions representing contrasting response levels was further inoculated with three additional isolates (Aph-1, AE11, and AE12). In this multi-isolate assay, disease severity was quantified, shoot biomass was recorded, and root system architecture traits were determined using WinRHIZO image analysis. A high correlation between foliar and root symptoms at 20 days indicated that foliar symptom assessment provides a reliable, non-destructive indicator of root health. Considerable variation in disease response was detected, with several genotypes maintaining consistently low symptom levels and three exhibiting near-complete resistance across all isolates. Root architectural traits further corroborated visual disease assessments, showing patterns consistent with resistance and susceptibility responses. Overall, this study demonstrates the presence of genetic variability in the response of V. sativa to A. euteiches, with a subset of accessions showing resistance to the four isolates tested. This resistance potential can be directly used in breeding programs focused on improving tolerance to root rot.

Legumes, due to their unique ability to fix atmospheric nitrogen, are a key factor in sustainable agriculture. However, like other crops, their production is affected by biotic and abiotic factors. Among them, Aphanomyces root rot is a significant threat in legume production globally. Peas and lentils manifest the most devastating consequences of this disease, but in the current environmentally changing scenario, similar consequences could be extended to other legume crops. Although comprehensive studies have been performed on Aphanomyces euteiches-legume pathosystem, the basis of their interaction is poorly understood due to its complexity and the highly diverse populations of this pathogen. In addition to these facts, the capacity of A. euteiches to adopt resting structures and to infect several legume species makes its management especially challenging. To date, only partially resistant accessions have been described, while chemical control effectiveness is limited to the seedling stage. Consequently, cultural practices, including crop rotations and determining infection levels prior to sowing, represent the sole reliable strategies nowadays. This review summarises the actual knowledge of A. euteiches-legume pathosystem as well as presents current strategies for its control and their limitations.

Pea (Pisum sativum L.) is an important temperate grain legume crop of high nutritional and agronomic value. Ascochyta blight, caused by a multi-species complex of necrotrophic fungi, remains a major constraint for pea production worldwide. This review synthesizes the available genetic, physiological and molecular knowledge on the pea-Ascochyta blight pathosystem, with emphasis on the genetic architecture of resistance, host defense mechanisms and the recent contributions from the omics disciplines. Current evidence indicates that genetic resistance to the various Ascochyta blight pathogens is incomplete and multicomponent, being associated with loci of small to moderate effect, with expression depending on organ, developmental stage and environment. Under field conditions, the observed phenotypes reflect the interaction between physiological resistance, plant architecture, phenology, canopy microenvironment and epidemic dynamics. Together, these factors bias phenotyping and limit the transferability of molecular markers. The practical value of these markers for use in marker-assisted selection (MAS) and genomic selection (GS) is presented and critically discussed. Future progress in breeding for Ascochyta blight resistance will depend on integrating molecular knowledge with a careful definition of ideotypes, well-calibrated phenotyping and multi-environment validation.

Anise (Pimpinella anisum L.) is one of the most important annual herbs of the Apiaceae family, widely cultivated in southern Spain. Their seeds are highly valued for culinary uses and for producing quality essential oils widely used in food and beverage products, as well as for industry, medicinal, and cosmetics applications. This study investigates the seed yield and essential oil content within a set of 50 anise accessions from worldwide origin, as well as their composition by GC–MS and GC–FID analysis. Accessions showed significant differences in the agronomic parameters measured, including plant height (cm), seed yield (kg ha−1), and the Harvest Index (%), with accessions PA_87 (Spain), PA_47 (Greece), and PA_21 (unknown origin) being the most performant. Essential oil (EO) content varied between 0.8% and 5.7% across different genotypes, resulting in EO production values ranging from 0.1 to 300 kg ha−1. Trans-anethole was identified as the dominant terpene, comprising 84.4% to 94.4% of the content, followed by eugenol (1.4% to 5.5%) and α-muurolene (1.4% to 7.2%). PCA analysis identified five distinct groups and one outlier, influenced by minor terpenes. Indeed, there was a strong negative correlation between estragole and pseudoisoeugenyl 2-methylbutyrate. This study underscores the significance of minor terpenes, which play crucial roles in defining unique aniseed chemotypes, allowing for the selection of cultivars optimized for specific uses in food, cosmetics, and pharmaceuticals. Additionally, these findings emphasize the impact of cultivar genetics on agronomic traits and EO profiles, suggesting the need for further research to optimize plant growth and yield and EO quality.

Context: Improving adaptation of field pea (Pisum sativum L.) to Mediterranean rainfed systems requires considering the combined effects of climate factors and key biotic constraints, including parasitic weeds like broomrape (Oc) and foliar diseases such as powdery mildew and Ascochyta blight. However, the combination of these stresses remain insufficiently understood. Objectives: To quantify and compare the performance, stability, and response to stresses of novel pea breeding lines; to identify agronomic and climatic predictors of yield; and to guide pea breeding for adaptation to Mediterranean rainfed systems. Methods: In this study, nine advanced pea breeding lines developed under the IAS-CSIC breeding programme were compared with five commercial cultivars in multi-environment trials across three seasons. Agronomic, phenological, and disease-related traits were assessed and analysed using Genotype & times;Environment (G & times;E) and multivariate approaches to address yield stability and to inform selection under biotic and abiotic stresses. Results: Among the biotic constraints, Oc had the strongest negative impact on grain yield. Higher levels of Oc parasitism were favoured by spring rainfall and high temperatures, thereby altering crop-environment interactions. IAS-CSIC breeding lines showed superior resistance to Oc, with approximately 50% fewer Oc shoots per plant than commercial cultivars. Consequently, they achieved higher yields in Oc-infested environments (1742 vs 743 kg ha-1). They also showed higher yields in Oc-free environments (3435 vs 2803 kg ha-1), highlighting consistent productivity and successful adaptation to Mediterranean rainfed systems. In Oc-free environments, abiotic-stress indices indicated that several breeding lines showed better yield maintenance than commercial cultivars under drier, hotter spring conditions. Rainfall showed beneficial or detrimental impacts on yield depending on the presence or absence of Oc. Several agronomic traits (i.e., plant height, crop appearance, lodging, and thermal time during pod filling) were consistent predictors of yield. The impact of powdery mildew on yield was limited under the studied conditions with several lines showing complete resistance. Multi-trait selection enabled the identification of breeding lines combining improved Oc resistance with yield stability and no trade-offs in agronomic traits or in response to other biotic stresses, particularly Cartujano, Chicana, and Pepapea. Conclusions and significance: Altogether, our results provide practical recommendations for growers in the Mediterranean region and methodological guidance for breeders. These findings illustrate the utility of combining resistance introgression, multi-trait selection, and trait-informed modelling as complementary strategies. This integrative framework may also support genetic improvement for adaptation to environments distinct from the original breeding context and exposed to diverse, and novel stresses.

Pea (Pisum sativum) production is challenged by drought stress. Traditional methods for assessing drought tolerance are limited, and high-throughput phenotyping (HTP) can facilitate the rapid and automated assessment of plant traits. Herein, 180 Pisum spp. accessions were evaluated using an indoor HTP platform under two irrigation treatments, control (70% field capacity) and drought stress (30% field capacity), for 50 days. A combination of digital phenotyping via imaging and manual measurements was used to analyse biomass-related, architectural, and physiological traits. Drought conditions resulted in significant reductions in biomass-related traits including fresh weight (47%), total leaf area (43%), and dry weight (41%). In contrast, PSII photochemical efficiency, leaf weight ratio, and solidity showed negative sensitivity index values (ranging from -7% to -1%), indicating comparatively lower sensitivity to drought and suggesting relative stability of these traits under water-limited conditions. The high heritability value for water use efficiency (0.87) suggests that this parameter may be useful for distinguishing pea's responses to suboptimal soil moisture levels. Principal component analysis (PCA) highlighted patterns of trait variation and associations among biomass-related traits, such as fresh weight, dry weight, and leaf area, which were sensitive to drought conditions. This suggests that the plants may use a combination of strategies to cope with water limitations. Furthermore, studying the significant variation in drought response among the diverse Pisum species and subspecies revealed distinct adaptation strategies. These findings support the development of crops that are resilient to the negative effects of climate change.

Ascochyta blight remains a major constraint for field pea (Pisum sativum L.) production and a priority for breeding programmes. So far, only moderate levels of incomplete resistance have been identified in pea germplasm and accumulated in pea cultivars by breeding. Resistance identified so far appears to be of complex inheritance, with phenotypic expression strongly affected by plant phenology and morphology and by environ-mental factors. This has slowed down the development and release of resistant elite cultivars. In this work, we describe the development of novel resistant breeding lines derived from targeted intra- and interspecific crosses combined with cycles of selection under high disease pressure at seedling and adult plant stages. The performance of thirteen breeding lines selected for improved resistance and good agronomic traits was further validated in a comparative field trial. Results confirmed the successful combination of competitive yield and good standing ability with good levels of resistance exceeding those of the resistant check. These advanced breeding lines are available on request for research and breeding use.

Agricultural diversification through species mixtures offers proven ecological and agronomic benefits, from improved nutrient cycling and yield stability to enhanced resilience against pests, diseases, and climatic stresses. Yet, the widespread uptake of these systems remains constrained by a fundamental gap: the absence of information on variety suitability for intercropping. Under current legislation, variety testing frameworks are designed for pure stand cultivation, thereby discouraging breeders from selecting within and for species mixtures, let alone registering mixture-adapted varieties. This Perspective article examines how current principles, policies, and procedures underpinning variety testing constrain the development and uptake of crop varieties suited for species mixtures. We synthesise evidence suggesting that integrating mixing ability into variety testing and registration could better align breeding incentives with farmer needs. Building on insights from a EU CAP Network Focus Group and long-standing forage and cereal-legume research, we outline conceptual and operational pathways to enable adaptation. A framework for action identifies key leverage points across five levels: breeding, regulation, farm networks, data infrastructures, and policy. Breeding must shift from isolated to interaction-based variety evaluation and selection, supported by traits and genomic predictors relevant for intercropping. When appropriate, official testing should add mixture sub-trials and recognise mixing ability as a sustainability trait, while linking practitioner networks and open data systems. Policy incentives and eco-labelling can help make diversity-oriented breeding economically viable. Rethinking breeding and variety testing in this way is pivotal for aligning breeding innovation, agricultural policy, and on-farm practice with the goals of resilient, diversified agroecosystems.

Abstract Pea (Pisum sativum L.) is an essential legume crop cultivated globally as food and feed. However, its production is greatly constrained by Fusarium oxysporum f. sp. pisi (Fop). Breeding for resistance is the most efficient management strategy, but the genetic foundation of Fop resistance remains unclear. Previous quantitative trait loci mapping has located several genomic regions associated with resistance to Fop. However, the large marker‐trait distances hampered the implementation of marker‐assisted selection. To unravel candidate genes for Fop races 1 and 2, diversity array technology (DArT) markers were applied to 324 pea core collections for a genome‐wide association study (GWAS). Phenotyping of the collections were performed under controlled growth chamber conditions with three independent experiments in a 324 × 3 × 7 factorial design. The collections were inoculated, and disease incidence was quantified over time using the area under the disease progress curve as the primary phenotypic measure for analyses. Phenotypic results revealed quantitative response and implicated wild accessions and landraces as favorable reservoirs of Fop resistance. GWAS using 26,045 DArT markers with three different models detected 15 marker‐trait associations (MTAs) for Fop race 1 and 27 MTAs for Fop race 2 resistance. MTAs were scattered across six pea chromosomes, with several of them located within the confidence interval of four previous Fop resistance loci—thereby refining their location. In addition, 28 potential candidate genes were identified near associated markers involved in Fop resistance, including genes encoding a reverse transcriptase, exocyst and conserved oligomeric Golgi complex subunits, a FERONIA‐like receptor kinase, homoserine kinase, a heat shock 70 protein, adenosine triphosphate transporters, and multiple transcription factors and plant defense‐related proteins. These putative genes and molecular pathways provide a foundation for the sustainable management of Fop.

Lathyrus sativus (grass pea) is a valuable crop for sustainable agriculture, offering dietary benefits and desirable agronomic traits. However, its yield stability is limited by diseases such as powdery mildew caused by Erysiphe pisi. Increasing fungal resistance to pesticides and environmental concerns demand the development of resistant crop varieties. To identify key defense mechanisms and effector genes involved in the Lathyrus sativus-Erysiphe pisi interaction we analyzed four L. sativus accessions exhibiting varying resistance to E. pisi (resistant, partially resistant, partially susceptible, and susceptible) using a dual RNA-Seq experiment across different time points. We observed a host biphasic response, characterized by an initial burst of gene expression, followed by a quiescent phase, and a subsequent wave of intense gene expression. Common L. sativus defense mechanisms included antifungal protein expression, cell wall reinforcement, and reactive oxygen species-mediated defense. These defenses involved respectively Bowman-Birk type proteinase inhibitors, peptidyl-prolyl cis-trans isomerases and mannitol dehydrogenases. The resistant accession specifically activated early reinforcement of structural barriers associated with lignin biosynthesis and the phenylpropanoid pathway, along with sustained chemical defenses (e.g. eugenol synthase 1), epigenetic regulation, and oxidative stress responses thorough peroxidases and heat shock proteins. The partial resistant accession exhibited a front-loaded defense response at early infection stages. Contrastingly, the partially susceptible accession exhibited a weaker baseline defense, with a slower and less robust response targeting pathogen infection. We identified potential E. pisi effectors, including genes involved in cell wall hydrolysis (e.g. mannosidase DCW1), nutrient acquisition (e.g. secreted alpha-glucosidase), and virulence (e.g. SnodProt1), with a higher diversity of effectors identified in the susceptible accession. In conclusion, this study identifies novel targets such as NLRs and effectors, antifungal proteins and genes related to cell wall reinforcement, within the complex Lathyrus sativus-Erysiphe pisi interaction to support future breeding programs aimed at enhancing resistance to E. pisi in L. sativus and related species.

Grass pea (Lathyrus sativus) is a nutrient-rich, and “climate-resilient” grain legume neglected due to low yields and the presence of the neurotoxin β-ODAP. Its crop wild relative, Lathyrus cicera, shows lower ODAP contents, earliness, and better adaptation to low-rainfall. Through sexual crossing, hybrids-derived lines were successfully developed and selected based on fertility and adaptation, but not thoroughly evaluated until the present study. This study evaluated 14 of these L. cicera x L. sativus hybrid-derived lines, alongside four parental lines, over three seasons in Mediterranean environments, assessing a total of 19 diverse traits. Traits assessed included growth-, cycle duration-, yield-, seed type-, pest susceptibility-, and seed quality-related. Linear mixed models were applied to analyze diversity and Principal Component Analysis summarized hybrids similarities. Selection and backcrossing resulted in introgressed plants resembling L. sativus parents in flower and seed coat colour, but for some traits were different from these parents. The introgression of L. cicera phenotypes was observed in cycle duration-related traits, with several hybrid-derived lines earlier than L. sativus parents, but similar or later than L. cicera parents. Likewise, for the quality-related trait L-homoarginine seed content, one hybrid-derived line depicted higher content than any parent, suggesting transgressive segregation. Transgressive segregation was also detected for some seed type-related traits, with hybrid-derived lines depicting longer or flatter seeds than any parent. The introgression of beneficial alleles from L. cicera into L. sativus background seems a reliable approach to create variability and combine desirable quality and adaptive traits for grass pea improvement under Mediterranean conditions.

Root rot caused by Aphanomyces euteiches is a major concern in pea (Pisum sativum L.). The lack of other effective control strategies makes crucial the development of resistant varieties. Although partial resistance has been reported, its quantitative inheritance, the association of resistance-linked genomic regions with unfavorable agronomic traits, and the limited understanding of soil pathogen populations hinder its progress in breeding programs. To search for alternative genomic regions associated with this partial resistance, a genome-wide association study (GWAS) was performed on a pea collection not yet explored for A. euteiches resistance in genetic studies. The 323 accessions of the collection were inoculated with RB84 isolate, and foliar and root symptoms were assessed 20 days after inoculation. The performed GWAS revealed 27 significantly associated markers among 26,045 SilicoDArT and 7033 single-nucleotide polymorphism marker datasets. Detected markers were distributed along the seven pea chromosomes, with 12 within previously described quantitative trait loci (QTLs). Chromosomes 2 and 5 harbored a significant number of associated markers, identified here for the first time, highlighting promising regions for future investigation. Twenty-one candidate resistance genes were identified. This study uncovers new genomic regions linked with A. euteiches resistance and provides molecular markers and candidate genes to support precision breeding. Newly identified QTL may be more effective against specific isolates than known QTL, enabling improved QTL rotation in the field.

The faba bean (Vicia faba) is an important grain legume that, despite decades of decline, is regaining interest in the Mediterranean basin due to an increasing demand for plant-based proteins and other ingredients, particularly for the food industry. However, the crop’s sensitivity to weather conditions (mainly drought and heat) as well as its high susceptibility to diseases hinder its yield performance and stability. For this reason, in this study, we present the results of multi-environment field trials conducted in southern Spain, where the performance of six new elite faba bean cultivars, developed through local breeding programs focused on selection for increased yield and chocolate spot (Botrytis fabae) resistance, was compared with two popular commercial cultivars. Data analysis across six diverse environments showed the significant effects of environment, genotype, and genotype-by-environment interaction (GEI) on yield and several morphologic traits. Grain yield was positively influenced by rainfall and negatively affected by high temperatures, with no evidence of damage due to cold temperatures. Stress tolerance indexes helped identify cultivars Omeya, Faraon Negro, and Navio6, which excelled across all metrics. The trials were intentionally conducted in broomrape (Orobanche crenata)-free plots, where chocolate spot emerged as the major biotic constraint, with the infection level highly influenced by rainfall. Significant differences were observed among accessions in their response to chocolate spot, with the cultivar Arrechana showing resistance. Overall, cultivars Omeya, Arrechana, Faraon Negro, Navio6, and Quijote demonstrated outstanding grain yield and excellent adaptation to the region.

Legumes contribute to sustainable agriculture by reducing fertilizer use, enhancing nitrogen fixation, and with high species diversity (~20,000 species). Spain is a leading EU producer, yielding up to 30,000 tons of different legume varieties annually. The Mediterranean climate, particularly in regions like Andalusia, is under increasing pressure from climate change, with extreme temperature variations and drought becoming more frequent. While these changes may jeopardize crop yields, limited information is available on their effects on the nutritional profile of legumes. From 2017 to 2019, six faba bean (Vicia faba) varieties were monitored in two climatically distinct areas of Andalusia to assess the impact of temperature (T) and rainfall (R) on key nutrients and bioactive compounds, including protein, minerals (K, Ca, Mg, Zn, P, Fe, Mn, B), total polyphenol content (TPC), tannins (TA), and saponins (S). Spearman correlations showed that higher T negatively impacted TPC (r = −0.40) and Mg (r = −0.33), while positively influencing Zn (r = 0.27) and Ca (r = 0.22). Rainfall increased TPC and Mg but reduced TA, Zn, and Ca. Canonical correspondence analysis (CCA) and PERMANOVA (p < 0.001) confirmed T, R, and yield as significant factors. These insights support breeding strategies for climate-adapted, nutrient-rich faba beans and the development of more resilient food systems.

Field pea seeds have long been recognized as valuable feed ingredients for animal diets, due to their high-quality protein and starch digestibility. However, the chemical composition of pea cultivars can vary across different growing locations, consequently impacting their nutrient profiles. This study employs untargeted metabolomics in conjunction with the quantification of fatty acids and amino acids to explore the influence of three different growing locations in Spain (namely Andalusia, Aragon and Asturias), on the nutritional characteristics of seeds of various pea cultivars. Significant interactions between cultivar and environment were observed, with 121 metabolites distinguishing pea profiles. Lipids, lipid-like molecules, phenylpropanoids, polyketides, carbohydrates, and amino acids were the most affected metabolites. Fatty acid profiles varied across locations, with higher C16:0, C18:0, and 18:1 n-9 concentration in Aragón, while C18:2 n-6 predominated in Asturias and C18:3 n-3 in Andalusia. Amino acid content was also location-dependent, with higher levels in Asturias. These findings underscore the impact of environmental factors on pea metabolite profiles and emphasize the importance of selecting pea cultivars based on specific locations and animal requirements. Enhanced collaboration between research and industry is crucial for optimizing pea cultivation for animal feed production.

Powdery mildew (Erysiphe pisi, E. trifolii) and rust (Uromyces pisi) are important diseases affecting grass pea (Lathyrus sativus). This study investigates grass pea's histological, enzymatic, and metabolic responses to these pathogens using accessions with contrasting resistance. Partially resistant (PR) accessions exhibited smaller fungal colonies from 48 h after inoculation (HAI) onwards. Enhanced superoxide dismutase (SOD) activity was observed as early as 12 HAI in PR accessions against both powdery mildews, associated with increased ascorbate peroxidase (APX) and catalase (CAT) activity in E. trifolii and E. pisi infections, respectively. Moreover, phenolic compounds and flavonoids accumulated in E. trifolii-infected PR accessions (6-48 HAI). For rust, APX activity rose at 48 HAI in PR accessions. These findings suggest that partial resistance (PR) in grass pea is characterized by restricted pathogen invasion and a dynamic regulation of reactive oxygen species (ROS)-scavenging enzymes, with responses varying across pathosystems. This highlights the importance of pathogen-specific selection strategies to minimize the risk of resistance breakdown and promote durable disease resistance in breeding programs.

GWAS using DArTseq markers identified novel resistance sources against parasitic broomrape in pea, elucidating candidate genes for marker-selected breeding as leverage for cultivar development and efficient disease control to enhance food security. Crenate broomrape (Orobanche crenata) is an important obligate root parasitic weed that causes severe yield losses in pea (Pisum sativum) production. O. crenata is difficult to eradicate in pea fields due to its high resilience and prolific seed boom capable of hibernating in soils for decades. Existing control strategies are not cost effective in low input legumes like pea. The most efficient ecofriendly mode of control is using resistant cultivars. Quantitative trait loci (QTL) studies based on bi-parental mapping has guided O. crenata resistance discovery, albeit their deployment in pea breeding is hindered by low marker resolution and large genetic distance. This study presents the first genome-wide association study (GWAS) on O. crenata resistance in pea, utilizing 324 diverse accessions and 26,045 diversity array technology sequence (DArTseq) markers. Phenotyping was performed over four seasons under field conditions using alpha lattice design. Results showed a strong phenotypic variation with an environmental influence on O. crenata infection. Novel resistance sources were identified mainly within the wild Pisum fulvum and P. sativum subsp. elatius. GWAS with two models yielded a total of 73 marker-trait associations with Chromosome 5 as major hotspot. Interestingly, some linked markers were detected in close proximity to four previous O. crenata resistance QTL. DArTseq markers identified 24 putative candidate genes participating in different cellular processes, including vesicle trafficking and transports, deoxyribonucleic acid transcription regulation, and defense including some leucine rich repeat receptor-like kinases. These results provide a valuable genetic resource for O. crenata resistance and a step toward its effective sustainable management—to enhance genetic diversity and cultivar improvement for food security.