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[EN] Short informative video about breeding activities on Lathyrus spp. and Pisum sativum at Córdoba (Spain) for the development of new crop varieties adaptated to the dryland Mediterranean conditions., [ES] Video divulgativo breve sobre las actividades de Mejora en Lathyrus spp. y Pisum sativum en Córdoba (España) para el desarrollo de nuevas variedades adaptadas al secano Mediterráneo.

Phenotyping data of legumes-cereals intercrops under field conditions at Córdob-Spain during two consecutive seasons 2021-2022 and 2022-2023.

Characterization data of a Vicia sativa germplams colection grown at Córdoba, Spain under three crop seasons.

Characterization data of a Pisum sp. germplams colection grown at Córdoba, Spain under three crop seasons

Characterization data of a Vicia faba germplams collection grown at Córdoba, Spain at three sites during 2022 and 2023.

[ES] Video divulgativo breve sobre lsa actividades de Mejora en Vicia sativa, Vicia faba y Pisum sativum en Córdoba, España., [EN] Short informative video about breeding activities on Vicia sativa, Vicia faba and Pisum sativum at Córdoba, Spain.

Fungal phytotoxins can be defined as secondary metabolites toxic to host plants and are believed to be involved in the symptoms developed of a number of plant diseases by targeting host cellular machineries or interfering with host immune responses. As any crop, legumes can be affected by a number of fungal diseases, causing severe yield losses worldwide. In this review, we report and discuss the isolation, chemical, and biological characterization of fungal phytotoxins produced by the most important necrotrophic fungi involved in legume diseases. Their possible role in plant-pathogen interaction and structure-toxicity relationship studies have also been reported and discussed. Moreover, multidisciplinary studies on other prominent biological activity conducted on reviewed phytotoxins are described. Finally, we explore the challenges in the identification of new fungal metabolites and their possible applications in future experiments.

Grain and forage legumes are important sources of food and feed, key for sustainable agriculture given the environmental services they provide. However, their cultivation is hampered in the Mediterranean Basin and Near East by the widespread occurrence of the root parasitic weed crenate broomrape (Orobanche crenata). Other broomrape species such as O. minor, O. foetida, and Phelipanche aegyptica are also of local importance. As for other parasitic weeds, a number of management strategies have been proposed, but considering that temperate legumes in the area are low-input crops, these strategies are largely uneconomical or hard to achieve, leaving the use of resistant cultivars as the most desirable option. Breeding for broomrape resistance is not an easy task, but significant progress has been achieved by classical breeding and selection and will profit from recent developments in phenomics and genomics. Here, achievements and prospects in broomrape management and resistance breeding are presented and critically discussed.

Legume-based cropping systems are regaining interest due to the environmental services and the variety of food and feed uses they provide, this having special interest in agroecological systems. There are many legume crops that can be adopted but focused breeding is needed to better meet the specific requirements of each system, especially in the situation of changing climate and often stressful environments. Standard breeding methods remain valid, from classical selection, to genomic assisted-breeding, profiting from the modern biotechnological and genomic approaches which are rapidly developing for most legume crops. Rather than focusing on the tools, emphasis is needed on the breeding targets, which might be different from those of high input agriculture such as improved response to symbiosis, nutrient and water use efficiency and tolerance to biotic and abiotic stresses becoming priorities. Breeding for these traits requires the infusion of genetic diversity from landraces or wild relatives by pre-breeding. Prospects and constraints are discussed.

Drought is increasingly frequent in the context of climate change and is considered a major constraint for crop yield. Water scarcity can impair growth, disturb plant water relations and reduce water use efficiency. Pea (Pisum sativum) is a temperate grain legume rich in protein, fibre, micronutrients and bioactive compounds that can benefit human health. In reducing pea yield because of drought, the intensity and duration of stress are critical. This review describes several drought resistance mechanisms in pea based on morphology, physiology and biochemical changes during/after the water deficit period. Drought tolerance of pea can be managed by adopting strategies such as screening, breeding and marker-assisted selection. Therefore, various biotechnological approaches have led to the development of drought-tolerant pea cultivars. Finally, the main objective of the current research is to point out some useful traits for drought tolerance in peas and also, mention the methods that can be useful for future studies and breeding programmes.

Figure S1. m/z distribution diagrams and Base Peak Chromatograms of C. truncatum isolate C428 (from lentil) extracts. (a) PDA extract; (b) PDB extract; (c) Rice extract; (d) Richard extract. Figure S2. m/z distribution diagrams and Base Peak Chromatograms of C. trucatum isolate C431 (from soybean) extracts. (a) PDA extract; (b) PDB extract; (c) Rice extract; (d) Richard extract. Figure S3. m/z distribution diagrams and Base Peak Chromatograms of C. trifolii isolate C436 (from clover extracts. (a) PDA extract; (b) PDB extract; (c) Rice extract; (d) Richard extract. Figure S4. Structure of: validated metabolites with pure standards (level A; red); putatively identified and produced by Colletotrichum spp. (level B(i), blue); putatively identified and produced by other fungal species (level B(ii), black); Structures of Curvupallide A and B (Level C(i), brown). Table S1. Parameters Metaboanalyst 5.0 for LC-MS spectra processing. Table S2. Features lists transformation and scaling for PLS – DA analysis. Q2 and R2 values of PLS – DA models in cross validation. Table S3. Metabolites dereplicated by targeted and untargeted metabolomics analysis, organised according to identification level. Table S4. Metabolites dereplicated by targeted and untargeted metabolomics analysis, organised according to identification level.

Lentil rust is a major disease worldwide caused by Uromyces viciae-fabae. In this study, we screened a large germplasm collection of cultivated lentils (Lens culinaris ssp. culinaris) and its wild relatives, both in adult plants in the field with a local rust isolate during 2 seasons and in seedlings under controlled conditions with four fungal isolates of worldwide origin. The main results from our study were the following: (1) a significant number of accessions with resistance based on hypersensitive reaction (reduced Infection Type (IT)) were identified in cultivated lentil and in L. ervoides, L. nigricans and L.c. orientalis. The IT scores showed a clear isolate-specific response suggesting race-specificity, so each fungal isolate might be considered a different race. Resistance was identified against all isolates what might be the basis to develop a standard differential set that should be a priority for rust definition and monitoring. (2) Interestingly, although at lower frequency than in L. ervoides and L. nigricans, the hypersensitive response was also observed within cultivated lentil, with accession 1561 (L.c. culinaris) displaying resistance to the four isolates making this accession a valuable ready-to-use resource for lentil resistance breeding. Resistance to all other rust isolates was also available within L.c. culinaris in an isolate-specific manner. Accession 1308 (L. ervoides) showed resistance against all isolates tested, as well as a reduced number of accessions belonging to other wild Lens species. (3) In addition, our screenings allowed the identification of several accessions with partial resistance (reduced Disease Severity (DS) despite high IT). Adult Plant Resistance resulting in reduced severity in adult plants in the field, despite high susceptibility in seedlings, was more frequently identified in L.c. culinaris, but also in L. nigricans and L.c. orientalis.

Supplementary Figure S1. 1H-NMR spectrum of ascochlorin (1) (CDCl3, 400 MHz); Supplementary Figure S2. ESI/MS (+) spectrum of ascochlorin (1); Supplementary Figure S3. 1H-NMR spectrum of ascofuranol (2) (CDCl3, 400 MHz); Supplementary Figure S4. ESI/MS (+) spectrum of ascofuranol (2); Supplementary Figure S5. 1H-NMR spectrum of (R)-mevalonolactone (3) (CDCl3, 400 MHz); Supplementary Figure S6. ESI/MS (+) spectrum of (R)-mevalonolactone (3); Supplementary Figure S7. 1H-NMR spectrum of ascosalipyrone (4) (CDCl3, 400 MHz); Supplementary Figure S8. ESI/MS (+) spectrum of ascosalipyrone (4); Supplementary Figure S9. 1H-NMR spectrum of benzoic acid (5) (CDCl3, 400 MHz); Supplementary Figure S10. ESI/MS (+) spectrum of benzoic acid (5); Supplementary Figure S11. 1H-NMR spectrum of tyrosol (6) (CDCl3, 400 MHz); Supplementary Figure S12. ESI/MS (+) spectrum of tyrosol (6); Supplementary Figure S13. 1H-NMR spectrum of ascosalitoxin (7) (CDCl3, 400 MHz); Supplementary Figure S14. ESI/MS (+) spectrum of ascosalitoxin (7); Supplementary Figure S15. Images of the symptoms of each of the compounds. Supplementary Table S1. Diseased area (mm2) measured on detached leaves of several legume crops with exudates from the fungus A. fabae growth in vitro on 3 different culture media (Czapek-Dox = CD, potato dextrose broth = PDB, and rice) at concentrations of 0.5, 1, and 2 mg/mL. Negative (blank untreated, water and MeOH 5%) controls were also included. The experiment was repeated four times. Supplementary Table S2. Diseased area (mm2) measured in leaves detached from various legume crops with metabolites produced by the exudate of the A. fabae fungus from the three-growth media at concentrations of 1, 10, and 100 µM. Negative controls (untreated blank, water, and MeOH 5%) were also included. p-value compared with value from MeOH 5% control.

Table S1: Response of the Lens germplasm accessions to U. viciae-fabae ex L. culinaris studied. DS = Disease Severity (%); IT = Infection Type according Stakman et al. (1962) [29], 6 where IT 0 = no symptoms, IT ; = necrotic flecks, IT 1 = minute pustules barely sporulating, IT 2 = necrotic halo surrounding small pustules, IT 3 = chlorotic halo and IT 4 = well-formed pustules with 7 no associated chlorosis or necrosis. 8 Ns = not studied

Peas (Pisum sativum) are the fourth most cultivated pulses worldwide and a critical source of protein in animal feed and human food. Developing pea core collections improves our understanding of pea evolution and may ease the exploitation of their genetic diversity in breeding programs. We carefully selected a highly diverse pea core collection of 325 accessions and established their genetic diversity and population structure. DArTSeq genotyping provided 35,790 polymorphic DArTseq markers, of which 24,279 were SilicoDArT and 11,511 SNP markers. More than 90% of these markers mapped onto the pea reference genome, with an average of 2787 SilicoDArT and 1644 SNP markers per chromosome, and an average LD50 distance of 0.48 and 1.38 Mbp, respectively. The pea core collection clustered in three or six subpopulations depending on the pea subspecies. Many admixed accessions were also detected, confirming the frequent genetic exchange between populations. Our results support the classification of Pisum genus into two species, P. fulvum and P. sativum (including subsp. sativum, arvense, elatius, humile, jomardii and abyssinicum). In addition, the study showed that wild alleles were incorporated into the cultivated pea through the intermediate P. sativum subsp. jomardii and P. sativum subsp. arvense during pea domestication, which have important implications for breeding programs. The high genetic diversity found in the collection and the high marker coverage are also expected to improve trait discovery and the efficient implementation of advanced breeding approaches.

Uromyces viciae-fabae is a highly specific biotrophic fungus that causes faba bean rust, one of the major diseases affecting this crop. We have assessed the feasibility of using intercropping (faba bean mixed with either pea, wheat or barley) or mixtures of susceptible and resistant cultivars to control rust both under field and controlled conditions. The results of four field intercropping experiments showed a significant reduction in rust severity on faba bean when intercropped with barley (average 22% reduction) but not with the other combinations. This reduction was also confirmed in studies under controlled conditions. The barrier effect of barley appears as the main mechanism explaining rust suppression. Additional experiments under controlled conditions showed that intercropping with barley did not influence the N content of faba bean and that different levels of N nutrition had no impact on rust severity in any case. The cultivar mixture field experiments showed that rust severity in the susceptible cultivar decreased as the proportion of the resistant cultivar in the mixture increased. The importance of the barrier effect of the resistant cultivars was determined in an experiment under controlled conditions. It can be concluded that crop diversification offers great potential to reduce rust in faba bean.

Assessing the adaptability and stability of herbicide-tolerant lentil accessions to two broad-spectrum post-emergence herbicides in multi-environment trials has become a must in a breeding program to improve its selection. The adaptability and stability of 42 herbicide-tolerant lentil accessions were investigated using five stability parameters under eight different environments. Significant Genotype–Environment (GE) interaction was found for days to flowering (DFLR), days to maturity (DMAT), and seed yield per plant (SY). The analyzed stability parameters such as Cultivar superiority, Finlay–Wilkinson, Shukla, Static Stability, and Wricke’s Ecovalence ranked the tested accessions differently, confirming the importance of using a combination of stability parameters when evaluating the performance of a group of accessions. GGE biplot of the SY trait accounted for 60.79% of sums of squares of the GE interaction and showed that cool and high rainfall environments are ideal for testing the agronomic performance of tolerant accessions. The GGE biplot of SY showed that IG4605(19), IG195(6), and IG156635(12) were specifically adapted to one mega environment, whereas IG70056(38) was identified as a superior line having a high and stable yield. These lines should be included in lentil crossing programs to develop herbicide-tolerant cultivars adapted to diverse environments.

Sulla is a biannual forage legume cultivated throughout the Mediterranean Basin. It can be severely damaged by powdery mildew, but there is little understanding on its causal species or its host range. The taxonomic characterization of Erysiphe species is mainly based on the examination of chasmothecia morphology, or on the molecular analysis of ITS1 and ITS2 sequences. However, a description of chasmothecia morphology or ITS sequences is not always available to clearly assign a given isolate to an existing or novel Erysiphe species. In an attempt to clarify the identity of the powdery mildew infecting sulla crop we studied the morphology and ITS of nine populations collected over years and countries and compared them with available ITS sequences. Phylogenetic analysis showed that the powdery mildews collected on sulla clustered together with isolates collected on Coronilla varia in an independent clade between the E. guarinonii/E. trifoliorum and E. palczewskii clades, strongly suggesting that all these sequences correspond to the same Erysiphe species. Measurements of conidia and conidial foot cells of the sulla-infecting powdery mildews partially overlap those of other Erysiphe species, although they clearly differentiate from them, supporting the assumption that powdery mildews from sulla belong to a distinct species as observed from the phylogenetic analysis. As far as we know, our study is the first to report the molecular characterization of powdery mildew isolated on plants from the Hedysarum clade. Cross-inoculations confirmed a high specialization of the powdery mildew of sulla, with the sulla’s isolates infecting only the S. coronaria accessions, and none of the accessions from the other legume genera studied. All studied S. coronaria accessions were heavily infected by the sulla isolate but not by any other isolate used. All this points to E. hedysari as the causal agent of the S. coronaria powdery mildew.

Multi-environment field testing of chickpea accessions winter sown in Southern Spain showed that environmental effects on yield were more important than genotypic effects and GEI. The most detrimental factor on grain yield was ascochyta blight infection. We did not find a significant effect of low temperatures on yield in the environments studied, probably due to the mild winters in the area. On the contrary, we found detrimental effects of high temperatures at the reproductive stage, particularly with numbers of days with Tmax >30 °C. We found that genotypic effects were larger than the environmental on ascochyta infection as we included accessions previously selected for their levels of resistance or susceptibility. Biplots based on the WAASB/productivity ratio highlighted AS19, AS30, AS23, AS26, and AS18 accessions as the best for productivity and stability of yield, matching with those with a lower ascochyta blight infection. The MTSI index also identified these as the best accessions for the region.

Pea rust is a major disease worldwide caused by Uromyces pisi in temperate climates. Only moderate levels of partial resistance against U. pisi have been identified so far in pea, urging for enlarging the levels of resistance available for breeding. Herein, we describe the responses to U. pisi of 320 Pisum spp. accessions, including cultivated pea and wild relatives, both under field and controlled conditions. Large variations for U. pisi infection response for most traits were observed between pea accessions under both field and controlled conditions, allowing the detection of genotypes with partial resistance. Simultaneous multi-trait indexes were applied to the datasets allowing the identification of partial resistance, particularly in accessions JI224, BGE004710, JI198, JI199, CGN10205, and CGN10206. Macroscopic observations were complemented with histological observations on the nine most resistant accessions and compared with three intermediates and three susceptible ones. This study confirmed that the reduced infection of resistant accessions was associated with smaller rust colonies due to a reduction in the number of haustoria and hyphal tips per colony. Additionally, a late acting hypersensitive response was identified for the first time in a pea accession (PI273209). These findings demonstrate that screening pea collections continues to be a necessary method in the search for complete resistance against U. pisi. In addition, the large phenotypic diversity contained in the studied collection will be useful for further association analysis and breeding perspectives.

E. pisi was thought to be the only causal agent of powdery mildew in peas, with three genes, er1, er2 and Er3, conferring resistance to this pathogen. Recently, E. trifolii has also been foundto cause this disease in peas in different countries, but its relevance in pea powdery mildew disease worldwide is unknown. The objective of this study was to develop a method to identify and quantify E. pisi and E. trifolii and use it to analyze the relative prevalence of E. pisi and E. trifolii in pea fields in Spain and Tunisia. We also wanted to discern the effect of the er1, er2 and Er3 resistance genes on the relative amount of E. pisi/E. trifolii. Using the polymorphic sites present between E. pisi and E. trifolii ITS sequences, we developed a qPCR method capable of identifying and quantifying these pathogens. Our results revealed, for the first time, the occurrence of E. trifolii in Tunisia and that the presence of er1, er2 and Er3 genes have a clear effect on the ratio E. pisi/E. trifolii in both countries.

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.