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Arsenic (As) is a toxic metalloid. As phyto-toxicity is manifested by its accumulation in different tissue types and subsequent growth inhibition in plants. Despite the vital role of leguminous crops in providing proteins to human diets, a little is known about the As accumulation in lentil. In this study, the rate of As uptake and transport from soil to root, shoot and grain of lentil as well as associated risks with the consumption of As contaminated food were examined. Biomass accumulation of lentil genotypes pardina, red chief and precoz drastically decreased when treated with As at 6 mg kg −1 concentration in comparison to 0 and 3 mg kg −1 As. Quantification of As concentrations following different treatment periods showed that As accumulation in roots and shoots of 0, 3 and 6 mg kg −1 As-treated lentil genotypes was statistically different. Arsenic content in grains of red chief genotype was found significantly lower than pardina and precoz. Moreover, As transport significantly increased in roots and shoots compared to the grains. Due to the high concentrations of As in biomass of lentil genotypes, animal as well as human health risk might be associated with the consumption of the As contaminated legume crops.

Background The membrane transporters viz. multidrug and toxic compound extrusion (MATE) and aluminum-activated malate transporter (ALMT) are associated with aluminum (Al) tolerance by accelerating secretion of organic acids, which can influence nutrient availability and stress response. However, such transporter families have not yet been reported in lentil under Al stress condition. Method and results In this study, 90 MATE and 14 ALMT genes were identified and clustered into four (MATE) and five (ALMT) subfamilies/clades with smaller subgroups. All the MATE and ALMT genes were unevenly dispersed across lentil chromosomes. Duplication analysis suggested that LcMATE gene family has expanded primarily through tandem duplication event. Collinearity of lentil with soybean suggested a close relationship between the MATE genes. The MATE promoter regions harboured many stress responsive as well as Al resistance transcription factor 1 related cis-regulatory elements. Predicted 3D (three-dimensional) structure and molecular docking revealed that 5 LcMATE proteins could bind citrate and contain amino acids related to its secretion via citrate exuding motif and other neighbouring sites. Expression analyses of LcMATE and LcALMT genes were performed using quantitative real-time polymerase chain reaction (qRT-PCR). Six genes namely, LcM1, LcM42, LcM46, LcM47, LcALMT8 and LcALMT14 responded to Al stress with varying levels of expression patterns at different time points (3, 6, 12 and 24 h). Conclusion Our findings offer thorough details on the MATE and ALMT transporters in lentils and will aid in valuable understanding for future functional studies of these transporters in generating Al tolerant cultivars.

Lentil (Lens culinaris ssp. culinaris) is a nutritious and affordable pulse with an ancient crop domestication history. The genus Lens consists of seven taxa, however, there are many discrepancies in the taxon and gene pool classification of lentil and its wild relatives. Due to the narrow genetic basis of cultivated lentil, there is a need towards better understanding of the relationships amongst wild germplasm to assist introgression of favourable genes into lentil breeding programs. Genotyping-by-sequencing (GBS) is an easy and affordable method that allows multiplexing of up to 384 samples or more per library to generate genome-wide single nucleotide Polymorphism (SNP) markers. In this study, we aimed to characterize our lentil germplasm collection using a two-enzyme GBS approach. We constructed two 96-plex GBS libraries with a total of 60 accessions where some accessions had several samples and each sample was sequenced in two technical replicates. We developed an automated GBS pipeline and detected a total of 266,356 genome-wide SNPs. After filtering low quality and redundant SNPs based on haplotype information, we constructed a maximum-likelihood tree using 5,389 SNPs. The phylogenetic tree grouped the germplasm collection into their respective taxa with strong support. Based on phylogenetic tree and STRUCTURE analysis, we identified four gene pools, namely L. culinaris/L. orientalis/L. tomentosus, L. lamottei/L. odemensis, L. ervoides and L. nigricans which form primary, secondary, tertiary and quaternary gene pools, respectively. We discovered sequencing bias problems likely due to DNA quality and observed severe run-to-run variation in the wild lentils. We examined the authenticity of the germplasm collection and identified 17% misclassified samples. Our study demonstrated that GBS is a promising and affordable tool for screening by plant breeders interested in crop wild relatives.

Soil salinity disrupts the physiological and biochemical processes of crop plants and ultimately leads to compromising future food security. Sodium nitroprusside (SNP), a contributor to nitric oxide (NO), holds the potential to alleviate abiotic stress effects and boost tolerance in plants, whereas less information is available on its role in salt-stressed lentils. We examined the effect of exogenously applied SNP on salt-stressed lentil plants by monitoring plant growth and yield-related attributes, biochemistry of enzymes (superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD)) amassing of leaf malondialdehyde (MDA) and hydrogen peroxide (H2O2). Salinity stress was induced by NaCl application at concentrations of 50 mM (moderate salinity) and 100 mM (severe salinity), while it was alleviated by SNP application at concentrations of 50 µM and 100 µM. Salinity stress severely inhibited the length of roots and shoots, the relative water content, and the chlorophyll content of the leaves, the number of branches, pods, seeds, seed yield, and biomass per plant. In addition, MDA, H2O2 as well as SOD, CAT, and POD activities were increased with increasing salinity levels. Plants supplemented with SNP (100 µM) showed a significant improvement in the growth- and yield-contributing parameters, especially in plants grown under moderate salinity (50 mM NaCl). Essentially, the application of 100 µM SNP remained effective to rescue lentil plants under moderate salinity by regulating plant growth and biochemical pathways. Thus, the exogenous application of SNP could be developed as a useful strategy for improving the performance of lentil plants in salinity-prone environments.

Enzymatic and antioxidative responses are key defense mechanisms in plants following pathogen invasion, collectively known as induced systemic resistance (ISR). Alternaria sp., a well-known soil-borne pathogen, causes blight diseases in various crops. This study investigates the defence response in lentil plants through the treatment-induced application of two multipotent pesticide degrading plant growth-promoting rhizobacteria (PGPR), Bacillus cereus and Bacillus safensis, to mitigate the destructive effects of Alternaria. Both bacterial strains were applied in different carrier-based bioformulations via soil drenching. We assessed the modulation of defense-related enzymes by various combinational treatments with the Alternaria pathogen. The in vitro production of antimicrobial compounds was analyzed using GC-MS to confirm their pathogen-suppressive capabilities. Field trials showed a positive correlation between treatments and improvements in yield and growth index (GI). The highest (180%) enzymatic induction of phenylalanine ammonia lyase (PAL) followed by catalase (CAT)(100%) and polyphenol oxidase (PPO) (54%), was observed in treatments with B. cereus alone or in combination with B. safensis, in presence of Alternaria, in respect to the control. In vitro analysis revealed the production of antimicrobial compounds, including benzoic acid derivatives, cyclotetrasiloxanes, hexacosane, chlorpyrifos, and phthalates, which may contribute to pathogen suppression. Our findings demonstrate that these biocontrol agents (BCAs) not only stimulate the plant's enzymatic defense system but also enhance growth, seed yield and produce several antimicrobial compounds in vitro. Thus, pesticide-tolerant PGPR, used in this study, exhibit both disease control and plant growth-promoting properties, offering promising applications in sustainable agriculture.

Domesticated lentil has a relatively narrow genetic base globally and most released varieties are susceptible to severe biotic and abiotic stresses. The crop wild relatives could provide new traits of interest for tailoring novel germplasm and cultivated lentil improvement. The primary objective of this study was to evaluate wild lentil accessions for identification of economically viable agro-morphological traits and resistance against major biotic stresses. The study has revealed substantial variations in seed yield and its important component characters. Further, the diversity analysis of wild accessions showed two major clusters which were bifurcated into sub-clusters, thereby suggesting their wider genetic divergence. However, principal component analysis exhibited that seed yield plant-1, number of seeds plant-1, number of pods plant-1, harvest index and biological yield plant-1 contributed significantly to the total genetic variation assessed in wild lentil taxa. Moreover, some of the wild accessions collected from Syria and Turkey regions showed resistance against more than one disease indicating rich diversity of lentil genetic resources. The identification of most promising genotypes carrying resistance against major biotic stresses could be utilized in the cultivated or susceptible varieties of lentil for enhancing genetic gains. The study has also identified some trait specific accessions, which could also be taken into the consideration while planning distant hybridization in lentil.

This study was conducted in order to test the effect of seed pretreatment or exogenous application through the rooting medium of 0.1 mM Salicylic Acid (SA) and 0.1 mM hydrogen peroxide (H 2 O 2 ) on growth, nutritional behavior and some biochemical parameters (photosynthetic pigments, gas exchange parameters, oxidative stress indicators and antioxidant enzymes activities) of lentil plants ( Lens culinaris ) under 75 mM salt stress. Our results demonstrated that salt stress noticeably reduced shoot and root DWs by 39.01 and 42.81%, respectively, as compared to controls. This reduction was associated with a significant decrease in all photosynthetic parameters, including Chlorophyll (Chl) and carotenoid (Car), net assimilation of photosynthesis ( A ), stomatal conductance ( gs ), transpiration ( E ) and internal CO 2 level ( Ci ), an accumulation of Na + and Cl - and a decrease of K + and Ca 2+ concentrations in plant shoots and roots. In addition, relative to control plants, salt stress remarkably increased the malondialdehyde MDA and H 2 O 2 contents especially in roots and increased GPOX and SOD activities, especially in plant shoots. Both methods of SA and H 2 O 2 application recovered the plant growth, enhanced shoot and root DWs (increase of 67.65 and 82.36% in shoots and roots, respectively, as compared to salt-stressed plants) and increased all parameters that were reduced by NaCl treatment. Nevertheless, the most prominent effects of SA and H 2 O 2 on plant growth were obtained with the seed priming method. Thus, SA and H 2 O 2 applications, especially the H 2 O 2 seed priming method, induced the antioxidant system, improved the membrane stability and ameliorated the gas exchange parameters. As compared to salt plant stressed, Na + and Cl - contents were significantly decreased and K + and Ca 2+ were significantly increased in shoots and roots following SA and H 2 O 2 applications, especially with the H 2 O 2 seed priming method. Similarly, this method was more efficient in alleviating the adverse effects of salt stress on all photosynthetic pigment contents and measured gas exchange parameters. Compared to salt stressed plants, it significantly decreased the H 2 O 2 and MDA contents and further stimulated GPOX and SOD activities. Our results indicated that the seed priming method, particularly with H 2 O 2 , could be recommended for obtaining better growth of lentil seedlings under salt-affected soil conditions.

Water being a vital part of cell protoplasm plays a significant role in sustaining life on earth; however, drastic changes in climatic conditions lead to limiting the availability of water and causing other environmental adversities. α-tocopherol being a powerful antioxidant, protects lipid membranes from the drastic effects of oxidative stress by deactivating singlet oxygen, reducing superoxide radicals, and terminating lipid peroxidation by reducing fatty acyl peroxy radicals under drought stress conditions. A pot experiment was conducted and two groups of lentil cultivar (Punjab-2009) were exposed to 20 and 25 days of drought induced stress by restricting the availability of water after 60 th day of germination. Both of the groups were sprinkled with α-tocopherol 100, 200 and 300 mg/L. Induced water deficit stress conditions caused a pronounced decline in growth parameters including absolute growth rate (AGR), leaf area index (LAI), leaf area ratio (LAR), root shoot ratio (RSR), relative growth rate (RGR), chlorophyll a, b, total chlorophyll content, carotenoids, and soluble protein content (SPC) which were significantly enhanced by exogenously applied α-tocopherol. Moreover, a significant increase was reported in total proline content (TPC), soluble sugar content (SSC), glycine betaine (GB) content, endogenous tocopherol levels, ascorbate peroxidase (APX), catalase (CAT) peroxidase (POD) and superoxide dismutase (SOD) activities. On the contrary, exogenously applied α-tocopherol significantly reduced the concentrations of malondialdehyde (MDA) and hydrogen peroxide (H 2 O 2 ). In conclusion, it was confirmed that exogenous application of α-tocopherol under drought induced stress regimes resulted in membrane protection by inhibiting lipid peroxidation, enhancing the activities of antioxidative enzymes (APX, CAT, POD, and SOD) and accumulation of osmolytes such as glycine betaine, proline and sugar. Consequently, modulating different growth, physiological and biochemical attributes.

Stemphylium blight (SB) is a major biotic stress that poses a potential threat to total yield loss in lentils. In this context, a comparative study was devised on the biochemical and physiological changes that may occur in six tolerant/resistant and six susceptible/highly susceptible lentil genotypes that were inoculated with the SB pathogen. Pathogen inoculation significantly reduced total chlorophyll and relative leaf water content across all genotypes, with the greatest reduction observed in susceptible genotypes. The recorded membrane stability index indicated significant membrane damage due to pathogen inoculation, particularly in susceptible genotypes. Similarly, the proline, total soluble protein, total soluble sugar, total phenol, and ascorbic acid contents increased several-fold in resistant genotypes when the pathogen was inoculated. Resistant lentil genotypes accumulated relatively high levels of antioxidant enzymes such as peroxidase, catalase, polyphenol oxidase, and phenyl ammonium lyase, while susceptible genotypes exhibited relatively low accumulation after pathogen inoculation. Pathogenesis-related enzymes like β- 1,3-glucanase, and chitinase were notably upregulated in the resistant genotypes compared to the susceptible genotypes. Fourteen biochemical parameters recorded from the screened genotype were subjected to principal component analysis (PCA). The results revealed that β- 1,3-glucanase, proline, ascorbic acid and chitinase activity were found to be excellent determinants of susceptibility or resistance of a particular genotype. In conclusion, it can be inferred that genotypes: G- 17 (7979/DPL62/2), G- 31 (8114/10956/16–1) and G- 13 (ILL10838/ILWL11/X2016S) exhibited favourable performance in terms of disease tolerance.

Background Lentil is a globally important pulse crop and a major dietary source of iron (Fe) and zinc (Zn). However, genotype × environment interaction (G × E) often leads to inconsistent micronutrient expression across environments, limiting genetic gain and the development of stable, nutrient-dense cultivars. Methods Twenty-five lentil genotypes, including three check varieties, were evaluated across seven environments using a randomised block design with three replications. Grain Fe and Zn concentrations were quantified using ICP–MS. Analysis of variance, AMMI, GGE biplot, and ASV (AMMI stability value) analyses were employed to assess G × E interaction, genotype stability, and adaptability. Results The AMMI model revealed that IPCA1 and IPCA2 were highly significant ( P  < 0.001), together explaining 85.7% of the variation for Fe and 88.2% for Zn. The AMMI model exhibits G7 and G13 to be stable genotypes for iron, while G23 shows general stability for zinc content. Based on ASV, G8, G13, and G12 were the most stable for Fe, whereas G23, G13, and G1 were the most stable for Zn. GGE biplot analysis showed that PC1 and PC2 accounted for 84.41% of the total variation in Fe and 82.54% in Zn. According to the GGE biplot, Genotype G13 and G3 exhibited superior performance for Fe, while G7, G24, and G21 were identified as high-performing for Zn. Three distinct mega-environments were identified, with each exhibiting different winning genotypes for iron, and three distinct mega-environments were identified with different winning genotypes for zinc. Conclusion Significant G × E interaction influences Fe and Zn accumulation in lentil. The identification of stable, high-micronutrient genotypes and distinct mega-environments underscores the importance of multi-environment testing. AMMI and GGE biplot analyses proved effective for dissecting G × E interaction and guiding the selection of nutritionally superior and stable lentil genotypes.