Salinity is one of the most severe environmental stresses, negatively affecting productivity of salt-sensitive crop species. Given that germination is the most critical phase in the plant life cycle, the present study aimed to determine seed germination potential and associated traits under salt stress conditions as a simple approach to identify salttolerant lentil genotypes.• The genetic material consisted of six lentil genotypes whose adaptation to various agroclimatic conditions is not well elucidated. Salinity stress was applied by addition of NaCl at three different levels of stress, while non-stressed plants were included as controls. Evaluation of tolerance was performed on the basis of germination percentage, seed water absorbance, root and shoot length, seedling water content, seedling vigour index and number of seedlings with an abnormal phenotype.• Overall, our findings revealed that salinity stress substantially affects all traits associated with germination and early seedling growth, with the effect of salinity being dependent on the level of stress applied. It is noteworthy, however, that genotypes responded differently to the varying salinity levels. In this context, Samos proved the most salt-tolerant genotype, indicating its possible use for cultivation under stress conditions.• In conclusion, the determination of seed germination and early growth potential may be exploited as an efficient strategy to reveal genetic variation in lentil germplasm of unknown tolerance to salinity stress. This approach allows selection of desirable genotypes at early growth stages, thus enabling more efficient application of various breeding methods to achieve stress-tolerant lentil genotypes.

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Among abiotic stresses, drought is undoubtedly one of the most severe environmental factors for a wide range of major crops, leading to considerable yield and economic losses. The adverse effects in crop yield reflect the result of a series of morphological and physiological changes but also changes in signaling pathways, transcriptional and post-transcriptional regulation of stress-responsive genes, and metabolic adaptations. Despite the exhausting studies elucidating plants’ metabolic response to drought, there is a knowledge gap in the biochemical mechanisms governing drought tolerance in lentil (Lens culinaris Medik.). The present study aimed to determine the fluctuations of the metabolite profiles of lentil genotypes with contrasting drought tolerance to discover possible biomarkers for screening tolerant genotypes at early growth stages. Lentil seedlings were subjected to osmotic drought stress, induced by polyethylene glycol, at two stress levels (2.5% and 5.0% PEG-6000) for a period of 20 days, while untreated plants were also included as controls. GC/ΕΙ/MS-mediated metabolic profiling was employed to monitor changes in response to osmotic drought stress. The data was subjected to OPLS-DA and OPLS-HCA for the discrimination between treatments and the discovery of trends and corresponding biomarkers. In total, the analysis yielded 150 metabolite features with highly reproducible patterns, of which the vast majority belonged to carbohydrates, carboxylic acids, and amino acids. Overall, findings highlight the differential accumulation of a series of compounds, and more importantly, the variable accumulation of certain metabolites, namely D-fructose, α,α-trehalose, myo-inositol, and L-tryptophan, in the contrasting genotypes, indicating that the adaptive metabolic responses to osmotic drought stress operate under strong genotypic dependency in lentil. Research findings provide insights into various aspects of lentil’s metabolism under drought and further offer the possibility of applying such knowledge towards effectively screening for drought-tolerant lentil germplasm at early growth stages.

BackgroundSpatial heterogeneity can have serious effects on the precision of field experimentation in plant breeding. In the present study the capacity of the honeycomb design (HD) to sample huge spatial heterogeneity was appraised. For this purpose, four trials were conducted each comprising a lentil landrace being screened for response to viruses.ResultsHuge spatial heterogeneity was reflected by the abnormally high values for coefficient of variation (CV) of single-plant yields, ranging 123–162 %. At a given field area, increasing the number of simulated entries was followed by declined effectiveness of the method, on account of the larger circular block implying greater intra-block heterogeneity; a hyperbolic increasing pattern of the top to bottom entry mean gap (TBG) indicated that a number of more than 100 replicates (number of plants per entry) is the crucial threshold to avoid significant deterioration of the sampling degree. Nevertheless, the honeycomb model kept dealing with variation better than the randomized complete block (RCB) pattern, thanks to the circular shape and standardized type of block that ensure the less possible extra heterogeneity with expanding the area of the block.ConclusionsOwing to the even and systematic entry allocation, breeders do not need to be concerned with the extra spatial heterogeneity that might induce the extra surface needed to expand the size of the block when many entries are considered. Instead, they could improve accuracy of comparisons with increasing the number of replicates (circular blocks) despite the concomitant greater overall spatial heterogeneity.

Previous efforts to transform lentil have been considerably hampered by the crop’s recalcitrant nature, giving rise to particularly low transformation and regeneration frequencies. This study aimed at optimizing an Agrobacterium rhizogenes-mediated transformation protocol for the generation of composite lentil plantlets, comprised of transgenic hairy roots and wild-type shoots. Transformation was performed by inoculating the cut hypocotyl of young lentil seedlings, while optimization involved the use of different bacterial strains, namely R1000, K599 and Arqua, and protocols differing in media composition with respect to the presence of acetosyringone and MES. Composite plantlets had a transgenic hairy root system characterized by an increased number of hairy roots at the hypocotyl proximal region, occasionally showing plagiotropic growth. Overall findings underline that transformation frequencies are subject to the bacterial strain, media composition as well as their combined effect. Among strains tested, R1000 proved to be the most capable of hairy root formation, while the presence of both acetosyringone and MES in inoculation and culture media yielded considerably higher transformation rates. The transgenic nature of hairy roots was demonstrated by the Ri T-DNA-mediated transfer of the rolB2 gene and the simultaneous absence of the virCD sequence of A. rhizogenes. Our findings provide strong evidence that A. rhizogenes-mediated transformation may be employed as a suitable approach for generating composite seedlings in lentil, a species whose recalcitrance severely hampers all efforts addressed to transformation and whole plant regeneration procedures. To the best of our knowledge, this is the first report on the development of a non-laborious and time-efficient protocol for the generation of transgenic hairy roots in lentil, thus providing an amenable platform for root biology and gene expression studies in the context of improving traits related to biotic and abiotic stress tolerance.