Identification of novel genes associated with herbicide tolerance in Lentil (Lens culinaris ssp. culinaris Medik.).

Weeds pose a major constraint in lentil cultivation, leading to decrease farmers' revenues by reducing the yield and increasing the management costs. The development of herbicide tolerant cultivars is essential to increase lentil yield. Even though herbicide tolerant lines have been identified in lentils, breeding efforts are still limited and lack proper validation. Marker assisted selection (MAS) can increase selection accuracy at early generations. Total 292 lentil accessions were evaluated under different dosages of two herbicides, metribuzin and imazethapyr, during two seasons at Marchouch, Morocco and Terbol, Lebanon. Highly significant differences among accessions were observed for days to flowering (DF) and maturity (DM), plant height (PH), biological yield (BY), seed yield (SY), number of pods per plant (NP), as well as the reduction indices (RI) for PH, BY, SY and NP. A total of 10,271 SNPs markers uniformly distributed along the lentil genome were assayed using Multispecies Pulse SNP chip developed at Agriculture Victoria, Melbourne. Meta-GWAS analysis was used to detect marker-trait associations, which detected 125 SNPs markers associated with different traits and clustered in 85 unique quantitative trait loci. These findings provide valuable insights for initiating MAS programs aiming to enhance herbicide tolerance in lentil crop.

Improvement of cool-season food legumes for adaptation to intercropping systems: breeding faba bean for intercropping with durum wheat as a case study.

Although the transition toward a more sustainable agricultural system is sparking the interest of scientists and farmers around the globe, breeding programs are still focusing on optimizing cultivars intended for the monoculture system, and most cultivars available on the market are not suitable for intercropping. The incorporation of versatile cool-season food legumes (CSFLs) in the intercropping system is a promising way toward more diversified and sustainable cropping systems. However, as the selection of good-performing cultivars under sole cropping does not always lead to a good performance in intercropping, the development of an alternative breeding scheme for intercropping is now a necessity. The case study of faba bean-wheat intercropping was used to select for traits associated with better performance of faba bean, resulting in identifying the combined grain yield, 100-seed weight, number of pods per plant, and canopy height as key traits for faba bean-wheat intercropping suitability. Incorporating these traits in the breeding programs would be the cornerstone of the prospective transition.

Metabolic Mechanisms Underlying Heat and Drought Tolerance in Lentil Accessions: Implications for Stress Tolerance Breeding.

Climate change has significantly exacerbated the effects of abiotic stresses, particularly high temperatures and drought stresses. This study aims to uncover the mechanisms underlying heat and drought tolerance in lentil accessions. To achieve this objective, twelve accessions were subjected to high-temperature stress (32/20 °C), while seven accessions underwent assessment under drought stress conditions (50% of field capacity) during the reproductive stage. Our findings revealed a significant increase in catalase activity across all accessions under both stress conditions, with ILL7814 and ILL7835 recording the highest accumulations of 10.18 and 9.33 under drought stress, respectively, and 14 µmol H2O2 mg protein-1 min-1 under high temperature. Similarly, ascorbate peroxidase significantly increased in all tolerant accessions due to high temperatures, with ILL6359, ILL7835, and ILL8029 accumulating the highest values with up 50 µmol ascorbate mg protein-1 min-1. In contrast, no significant increase was obtained for all accessions subjected to water stress, although the drought-tolerant accessions accumulated more APX activity (16.59 t to 25.08 µmol ascorbate mg protein-1 min-1) than the sensitive accessions. The accessions ILL6075, ILL7814, and ILL8029 significantly had the highest superoxide dismutase activity under high temperature, while ILL6363, ILL7814, and ILL7835 accumulated the highest values under drought stress, each with 22 to 25 units mg protein-1. Under both stress conditions, ILL7814 and ILL7835 recorded the highest contents in proline (38 to 45 µmol proline/g FW), total flavonoids (0.22 to 0.77 mg QE g-1 FW), total phenolics (7.50 to 8.79 mg GAE g-1 FW), total tannins (5.07 to 20 µg CE g-1 FW), and total antioxidant activity (60 to 70%). Further, ILL7814 and ILL6338 significantly recorded the highest total soluble sugar content under high temperature (71.57 and 74.24 mg g-1, respectively), while ILL7835 achieved the maximum concentration (125 mg g-1) under drought stress. The accessions ILL8029, ILL6104, and ILL7814 had the highest values of reducing sugar under high temperature with 0.62 to 0.79 mg g-1, whereas ILL6075, ILL6363, and ILL6362 accumulated the highest levels of this component under drought stress with 0.54 to 0.66 mg g-1. Overall, our findings contribute to a deeper understanding of the metabolomic responses of lentil to drought and heat stresses, serving as a valuable reference for lentil stress tolerance breeding.

Impact of Heat and Drought Stress on Grasspea and Its Wild Relatives.

Grasspea (Lathyrus sativus L.) is recognized as a highly drought-tolerant legume. However, excessive consumption of its seeds and green tissues causes neurolathyrism, a condition characterized by an irreversible paralysis of the legs induced by a neurotoxin amino acid called ß-N-oxalyl-L-α, ß- diaminopropionic acid (ß-ODAP). The present study investigated the effects of heat, and combined heat + drought during the reproductive phase on physiological and phenological parameters, yield-related factors, ODAP content, and seed protein of 24 genotypes representing 11 Lathyrus species under controlled conditions. Analysis of variance revealed a highly significant effect (p < 0.001) of stress treatments and genotypes for all the traits. In general, heat stress individually or in combination with drought expedited phenology, reduced relative leaf water content, stimulated proline synthesis, and influenced chlorophyll concentration; the effects were more severe under the combined heat + drought stress. ODAP content in seeds ranged from 0.06 to 0.30% under no-stress conditions. However, under heat stress, there was a significant increase of 33% in ODAP content, and under combined stress (heat + drought), the increase reached 83%. Crude protein content ranged from 15.64 to 28.67% among no stress plants and decreased significantly by 23% under heat stress and by 36% under combined stress. The findings of this study also indicated substantial reductions in growth and grain yield traits under both heat stress and combined heat + drought stress. Six accessions namely IG 66026, IG 65018, IG 65687, IG 118511, IG 64931, and IG65273 were identified as having the most favorable combination of yield, protein content, and seed ODAP levels across all conditions. ODAP content in these six accessions varied from 0.07 to 0.11% under no stress and remained at moderate levels during both heat stress (0.09-0.14%) and combined stress (0.11-0.17%). IG 66026 was identified as the most stable genotype under drought and heat stress conditions with high protein content, and low ODAP content. By identifying those promising accessions, our results have established a basis for forthcoming grasspea breeding initiatives while paving the way for future research exploration into the fundamental mechanisms driving ODAP variation in the presence of both heat and drought stress conditions.

Assessing the Stability of Herbicide-Tolerant Lentil Accessions (Lens culinaris Medik.) under Diverse Environments.

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.

Effect of High Temperature Stress During the Reproductive Stage on Grain Yield and Nutritional Quality of Lentil (Lens culinaris Medikus).

High temperature during the reproductive stage limits the growth and development of lentil (Lens culinaris Medikus). The reproductive and seed filling periods are the most sensitive to heat stress, resulting in limited yield and nutritional quality. Climate change causes frequent incidents of heat stress for global food crop production. This study aimed to assess the impact of high temperature during the reproductive stage of lentil on grain yield, nutritional value, and cooking quality. Thirty-six lentil genotypes were evaluated under controlled conditions for their high temperature response. Genotypic variation was significant (p < 0.001) for all the traits under study. High temperature-induced conditions reduced protein, iron (Fe) and zinc (Zn) concentrations in lentils. Under heat stress conditions, mineral concentrations among lentil genotypes varied from 6.0 to 8.8 mg/100 g for Fe and from 4.9 to 6.6 mg/100 g for Zn. Protein ranged from 21.9 to 24.3 g/100 g. Cooking time was significantly reduced due to high temperature treatment; the range was 3-11 min, while under no stress conditions, cooking time variation was from 5 to 14 min. Phytic acid variation was 0.5-1.2 g/100 g under no stress conditions, while under heat stress conditions, phytic acid ranged from 0.4 to 1.4 g/100 g. All genotypes had highly significant bioavailable Fe and moderately bioavailable Zn under no stress conditions. Whereas under heat stress conditions, Fe and Zn bioavailability was reduced due to increased phytic acid levels. Our results will greatly benefit the development of biofortified lentil cultivars for global breeding programs to generate promising genotypes with low phytic acid and phytic acid/micronutrient ratio to combat micronutrient malnutrition.

Genetic Dissection of Heat Stress Tolerance in Faba Bean (Vicia faba L.) Using GWAS.

Heat waves are expected to become more frequent and intense, which will impact faba bean cultivation globally. Conventional breeding methods are effective but take considerable time to achieve breeding goals, and, therefore, the identification of molecular markers associated with key genes controlling heat tolerance can facilitate and accelerate efficient variety development. We phenotyped 134 accessions in six open field experiments during summer seasons at Terbol, Lebanon, at Hudeiba, Sudan, and at Central Ferry, WA, USA from 2015 to 2018. These accessions were genotyped using genotyping by sequencing (GBS), and 10,794 high quality single nucleotide polymorphisms (SNPs) were discovered. These accessions were clustered in one diverse large group, although several discrete groups may exist surrounding it. Fifteen lines belonging to different botanical groups were identified as tolerant to heat. SNPs associated with heat tolerance using single-trait (ST) and multi-trait (MT) genome-wide association studies (GWASs) showed 9 and 11 significant associations, respectively. Through the annotation of the discovered significant SNPs, we found that SNPs from transcription factor helix-loop-helix bHLH143-like S-adenosylmethionine carrier, putative pentatricopeptide repeat-containing protein At5g08310, protein NLP8-like, and photosystem II reaction center PSB28 proteins are associated with heat tolerance.

Experimental on-farm trials data of faba bean and wheat intercropping field validation in Lebanon and Morocco.

This data paper describes the content of four datasets collected by the International Center for Agricultural Research in the Dry Areas (ICARDA) as a partner in the project "Designing InnoVative plant teams for Ecosystem Resilience and agricultural Sustainability (DIVERSify)" with the objective of assessing the feasibility of faba bean-wheat mixture in Mediterranean environments under diverse rainfed conditions. Data was collected during the trials conducted in Kfardan-Lebanon during 2017/2018 where 40 faba bean varieties were evaluated as sole and as mixture with 2 wheat cultivars 'Margherita' and 'Miki' and during 2018/2019 where 40 faba bean varieties and one durum wheat cultivar 'Margherita' were evaluated under low rainfall environments. Trials were also conducted in Tal Amara-Lebanon during 2019/2020 where 20 faba bean lines and one durum wheat cultivar 'Margherita' were evaluated under high rainfall environments and in Marchouch-Morocco during 2019/2020 where 7 faba bean lines with 3 cultivars and one durum wheat cultivar 'Margherita' were evaluated under extremely low rainfall environments. A detailed list of the different biological traits collected for wheat and faba bean is found in the specification table in this article. The Kfardan 2018/ 2019, Tal Amara and Marchouch data is related to the conference paper "Performance of faba bean-wheat mixture under diverse Mediterranean environments" [1].

Adaptability and Stability of Faba Bean (Vicia faba L.) Accessions under Diverse Environments and Herbicide Treatments.

The adaptability and stability of 37 faba bean (Vicia faba L.) accessions with different levels of tolerance to metribuzin or imazethapyr was assessed across 12 season-location-herbicide experiments. Significant Genotype x environment (GE) interaction was found for the days to flowering (DFLR), plant height (PLHT) and grain yield (GY). Performance and stability of the accessions regarding PLHT and GY were assessed using four different stability parameters: cultivar superiority, static stability, Wricke's eco-valence and Finlay and Wilkinson's regression model. The stability parameters ranked these genotypes differently suggesting that PLHT and GY stability should be assessed not only on a single or a few stability parameters but on a combination of them. GGE biplot analysis indicated that the environments representing metribuzin treatment at Marchouch 2014-2015 and the non-treated treatment at Terbol 2018-2019 are the ideal environments for evaluating faba bean genotypes. GGE biplots showed herbicide tolerant accession IG12983 with simultaneous average PLHT, GY and stability across the environments. The performance of other tolerant accessions, namely IG13945, IG13906, IG106453, FB2648, and FB1216 was less stable but superior under specific mega environments. Therefore, utilizing these accessions in faba bean breeding programs would help broaden the adaptability to diverse locations-season-herbicide treatments.

Genomic regions associated with herbicide tolerance in a worldwide faba bean (Vicia faba L.) collection.

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.