Lentil Genome Research Articles

Omics-Driven Strategies for Developing Saline-Smart Lentils: A Comprehensive Review

A number of consequences of climate change, notably salinity, put global food security at risk by impacting the development and production of lentils. Salinity-induced stress alters lentil genetics, resulting in severe developmental issues and eventual phenotypic damage. Lentils have evolved sophisticated signaling networks to combat salinity stress. Lentil genomics and transcriptomics have discovered key genes and pathways that play an important role in mitigating salinity stress. The development of saline-smart cultivars can be further revolutionized by implementing proteomics, metabolomics, miRNAomics, epigenomics, phenomics, ionomics, machine learning, and speed breeding approaches. All these cutting-edge approaches represent a viable path toward creating saline-tolerant lentil cultivars that can withstand climate change and meet the growing demand for high-quality food worldwide. The review emphasizes the gaps that must be filled for future food security in a changing climate while also highlighting the significant discoveries and insights made possible by omics and other state-of-the-art biotechnological techniques.

Advancements in Lentil Genomics for Enhanced Crop Breeding: A Review

Lentil (Lens culinaris Medik) is an essential pulse crop that is widely grown for its high nutritional value, notably its high protein content, making it an important dietary component for vegetarians and vegans. Despite being the world's fifth most produced pulse, with large contributions from Canada and India, lentil production confronts obstacles such as poor productivity due to limited genetic improvement against biotic and abiotic stresses under rainfed cultivation conditions. Recent advances in lentil genetics and genomics, such as the discovery of genes related to yield, disease resistance, and nutritional content, have boosted breeding efforts to generate improved lentil varieties. The use of contemporary genomic techniques like molecular markers, marker-assisted selection (MAS), genomic selection (GS), and next-generation sequencing (NGS) technology has sped up the discovery of quantitative trait loci (QTLs) and the production of novel cultivars with superior agronomic characteristics. Databases such as NCBI and ENA, as well as specialized resources like KnowPulse, provide critical genomic data, while the creation of lentil genome assemblies, notably the CDC Redberry variety, has improved our understanding of lentil genetics. These resources help to solve the constraints of traditional breeding, particularly for complex characteristics impacted by genotype-environment interactions, opening the way for more robust and productive lentil varieties. Although the application of advanced tools such as genetic engineering, cisgenesis, and genome editing has moved more slowly in lentils than in other crops, their potential to improve lentil output is encouraging. Recent studies on lentil genomes, together with the creation of increased genetic resources and cutting-edge techniques, offer the ability to overcome production constraints and dramatically increase lentil production and quality throughout the world.

INTEGRATION OF NOVEL SSR MARKERS INTO THE LENTIL (Lens culinaris Medik.) GENOME

The development of Simple Sequence Repeat markers (SSRs) for lentil has played a pivotal role in enhancing the comprehension of the lentil genome through genetic mapping. The study aimed to determine the relative positions of newly developed microsatellites to the lentil genome using an F7-derived Recombinant Inbred Lines population (RIL) of 71 individuals developed from a cross between Eston and PI320937. Molecular analysis was performed with 100 newly developed lentil SSR markers and a linkage map was constructed using MapMaker/EXP 3.0b and MapChart 2.2 software. Among the 100 SSR markers, 12 markers exhibited polymorphism, 54 markers were identified as monomorphic, and 34 markers remained unamplified. While 10 out of the 12 polymorphic markers successfully integrated into two linkage groups, covering a cumulative length of 19.2cM, two markers remained unlinked. Linkage group-1, comprised of 8 markers, spanned 4.8cM, and linkage group-2 extended over a length of 14.4cM with two markers. Despite only partially representing 2 out of the 7 chromosomes in the lentil genome, this map holds promise for future mapping studies. Through the addition of markers, it could facilitate marker-assisted selection and the identification of QTLs associated with specific agronomic traits.

Genome wide identification and expression profiling of ATP binding cassette (ABC) transporters gene family in lentil (Lens culinaris Medikus) under aluminium stress condition

Adenosine triphosphate-binding cassette transporters (ABC transporters) are involved in regulating plant growth, development and tolerance to environmental stresses. In this study, a total of 138 ABC transporter genes were identified in the lentil genome that were classified into eight subfamilies. Four lentil ABC transporters from subfamily B and I were clustered together with the previously characterized ABC transporter proteins related to aluminium (Al) detoxification. Lentil ABC transporter genes were distributed across the chromosomes. Tandem duplication was the main driving force for expansion of the ABC gene family. Collinearity of lentil with soybean indicated that ABC gene family is closely linked to Glycine max. ABC genes in the same subfamily showed similar gene structure and conserved motifs. The ABC promoter regions harboured a large number of plant hormones and multiple stress responsive cis-regulatory elements. The qRT-PCR showed that ABC genes had varied expression in roots of lentil at different time points under Al stress. This is the first report on genome wide identification and expression analyses of genes encoding ABC transporter genes in lentil which has provided in-depth insight for future research on evolution and elucidation of molecular mechanisms for aluminium tolerance.

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.

Genome-wide discovery of InDels and validation of PCR-Based InDel markers for earliness in a RIL population and genotypes of lentil (Lens culinaris Medik.).

The systematic identification of insertion/deletion (InDel) length polymorphisms from the entire lentil genome can be used to map the quantitative trait loci (QTL) and also for the marker-assisted selection (MAS) for various linked traits. The InDels were identified by comparing the whole-genome resequencing (WGRS) data of two extreme bulks (early- and late-flowering bulk) and a parental genotype (Globe Mutant) of lentil. The bulks were made by pooling 20 extreme recombinant inbred lines (RILs) each, derived by crossing Globe Mutant (late flowering parent) with L4775 (early flowering parent). Finally, 734,716 novel InDels were identified, which is nearly one InDel per 5,096 bp of lentil genome. Furthermore, 74.94% of InDels were within the intergenic region and 99.45% displayed modifier effects. Of these, 15,732 had insertions or deletions of 20 bp or more, making them amenable to the development of PCR-based markers. An InDel marker I-SP-356.6 (chr. 3; position 356,687,623; positioned 174.5 Kb from the LcFRI gene) was identified as having a phenotypic variance explained (PVE) value of 47.7% for earliness when validated in a RIL population. Thus, I-SP-356.6 marker can be deployed in MAS to facilitate the transfer of the earliness trait to other elite late-maturing cultivars. Two InDel markers viz., I-SP-356.6 and I-SP-383.9 (chr. 3; linked to LcELF3a gene) when tested in 9 lentil genotypes differing for maturity duration, clearly distinguished three early (L4775, ILL7663, Precoz) and four late genotypes (Globe Mutant, MFX, L4602, L830). However, these InDels could not be validated in two genotypes (L4717, L4727), suggesting either absence of polymorphism and/or presence of other loci causing earliness. The identified InDel markers can act as valuable tools for MAS for the development of early maturing lentil varieties.

Molecular characterization of acetolactate synthase genes in lentil (Lens culinaris Medik.): A key target enzyme of imazethapyr herbicide resistance

Weed infestation severely affects the productivity of lentils. Imazethapyr, a Group 2 herbicide controls prominent broadleaf weeds and grasses in the field. Imazethapyr blocks the first step of branched-chain amino acids (BCAA) biosynthesis by inhibiting the activity of acetolactate synthase (ALS) activity. The present study aims to explore the effect of herbicide stress on the ALS gene expression and enzyme activity in lentil genotypes on different days after spray. The study revealed the presence of two homologous genes in the lentil genome, LcALS2 and LcALS3, present on chromosomes 2 and 3, respectively. qRT-PCR of the LcALS2 gene in the selected herbicide-tolerant (LL1397 and LL1612) and susceptible (FLIP 2004-7 L and PL07) genotypes indicated a significant decrease of the ALS gene expression in the susceptible genotypes. The results were also supported by the in vitro ALS enzyme activity assay, indicating significant inhibition of ALS activity in FLIP 2004-7 L (17.33% and 49.21%) followed by PL07 (33.11% and 19.7%) at 4 and 7 days after spray (DAS), respectively compared to the control. This is the first-ever report on the effect of herbicide stress on ALS gene expression and its correlation with ALS enzyme activity on different days after spraying in lentil crop. It is important from an agronomic perspective to understand herbicide tolerance mechanism at the genetic level for selecting an efficient variety and improving crop productivity.

Next-Generation-Sequencing-Based Simple Sequence Repeat (SSR) Marker Development and Linkage Mapping in Lentil (Lens culinaris L.).

Simple sequence repeats (SSRs) are highly versatile markers in genetic diversity analysis and plant breeding, making them widely applicable. They hold potential in lentil (Lens culinaris) breeding for genetic diversity analysis, marker-assisted selection (MAS), and linkage mapping. However, the availability and diversity of SSR markers in lentil is limited. We used next-generation sequencing (NGS) technology to develop SSR markers in lentil. NGS allowed us to identify regions of the lentil genome that contained SSRs. Illumina Hiseq-2000 sequencing of the lentil genotype "Karacadağ" resulted in 1,727,734 sequence reads comprising more than 48,390 Mb, and contigs were mined for SSRs, resulting in the identification of a total of 8697 SSR motifs. Among these, dinucleotide repeats were the most abundant (53.38%), followed by trinucleotides (30.38%), hexanucleotides (6.96%), tetranucleotides (6.59%), and pentanucleotides (3.19%). The most frequent repeat in dinucleotides was the TC (21.80%), followed by the GA (17.60%). A total of 2000 primer pairs were designed from these motifs, and 458 SSR markers were validated following their amplified PCR products. A linkage map was constructed using these new SSRs with high linkage disequilibrium (209) and previously known SSRs (11). The highest number of SSR markers (43) was obtained in LG2, while the lowest number of SSR markers (19) was obtained in LG7. The longest linkage group (LG) was LG2 (86.84 cM), whereas the shortest linkage group was LG7 (53.46 cM). The average length between markers ranged from 1.86 cM in LG1 to 2.81 cM in LG7, and the map density was 2.16 cM. The developed SSRs and created linkage map may provide useful information and offer a new library for genetic diversity analyses, linkage mapping studies, and lentil breeding programs.

Identification of key genes and molecular mechanisms associated with temperature stress in lentil.

Extreme temperature is one of the serious threats to crop production in present and future scenarios of global climate changes. Lentil (Lens culinaris) is an important crop, and there is a serious lack of genetic information regarding environmental and temperature stresses responses. This study is the first report of evaluation of key genes and molecular mechanisms related to temperature stresses in lentil using the RNA sequencing technique. De novo transcriptome assembly created 44,673 contigs and differential gene expression analysis revealed 7494 differentially expressed genes between the temperature stresses and control group. Basic annotation of generated transcriptome assembly in our study led to the identification of 2765 novel transcripts that have not been identified yet in lentil genome draft v1.2. In addition, several unigenes involved in mechanisms of temperature sensing, calcium and hormone signaling and DNA-binding transcription factor activity were identified. Also, common mechanisms in response to temperature stresses, including the proline biosynthesis, the photosynthetic light reactions balancing, chaperone activity and circadian rhythms, are determined by the hub genes through the protein-protein interaction networks analysis. Deciphering the mechanisms of extreme temperature tolerance would be a new way for developing crops with enhanced plasticity against climate change. In general, this study has identified set of mechanisms and various genes related to cold and heat stresses which will be useful in better understanding of the lentil's reaction to temperature stresses.

Characterization of Genetic and Allelic Diversity Amongst Cultivated and Wild Lentil Accessions for Germplasm Enhancement.

Intensive breeding of cultivated lentil has resulted in a relatively narrow genetic base, which limits the options to increase crop productivity through selection. Assessment of genetic diversity in the wild gene pool of lentil, as well as characterization of useful and novel alleles/genes that can be introgressed into elite germplasm, presents new opportunities and pathways for germplasm enhancement, followed by successful crop improvement. In the current study, a lentil collection consisting of 467 wild and cultivated accessions that originated from 10 diverse geographical regions was assessed, to understand genetic relationships among different lentil species/subspecies. A total of 422,101 high-confidence SNP markers were identified against the reference lentil genome (cv. CDC Redberry). Phylogenetic analysis clustered the germplasm collection into four groups, namely, Lens culinaris/Lens orientalis, Lens lamottei/Lens odemensis, Lens ervoides, and Lens nigricans. A weak correlation was observed between geographical origin and genetic relationship, except for some accessions of L. culinaris and L. ervoides. Genetic distance matrices revealed a comparable level of variation within the gene pools of L. culinaris (Nei’s coefficient 0.01468–0.71163), L. ervoides (Nei’s coefficient 0.01807–0.71877), and L. nigricans (Nei’s coefficient 0.02188–1.2219). In order to understand any genic differences at species/subspecies level, allele frequencies were calculated from a subset of 263 lentil accessions. Among all cultivated and wild lentil species, L. nigricans exhibited the greatest allelic differentiation across the genome compared to all other species/subspecies. Major differences were observed on six genomic regions with the largest being on Chromosome 1 (c. 1 Mbp). These results indicate that L. nigricans is the most distantly related to L. culinaris and additional structural variations are likely to be identified from genome sequencing studies. This would provide further insights into evolutionary relationships between cultivated and wild lentil germplasm, for germplasm improvement and introgression.

Widening the genetic base of cultivated gene pool following introgression from wild Lens taxa

AbstractWild Lens taxa are a reservoir of useful rare genes/alleles for widening the genetic base and synthesis of a new gene pool of lentil. To maximize and sustain lentil production, new gene sources are needed to be identified and incorporated into cultivated background. This needs a comprehensive approach to accumulate favourable alleles from distantly related germplasm for widening of the cultivated gene pool and would be the most appropriate strategy to solve the various problems associated with stressed crop production and plateaued yields. Furthermore, expansion of deeper understanding of lentil genomics along with extensive research undertaken in other crop species can provide suitable guidelines to cover the distribution of Lens genus and component gene pools for further remarkable progress in lentil genetic improvement. This review aims at the genus Lens distribution and gene pools, crop germplasm conserved in ex‐situ and in‐situ collection, wild species characterization and evaluation for useful traits of interest to solve production‐related problems, highlight useful gene sources present in different gene pools and the progress achieved for widening the genetic base of cultivated varieties of lentil through wide hybridization and exploring lentil genomics.

A consensus linkage map of lentil based on DArT markers from three RIL mapping populations.

BackgroundLentil (Lens culinaris ssp. culinaris Medikus) is a diploid (2n = 2x = 14), self-pollinating grain legume with a haploid genome size of about 4 Gbp and is grown throughout the world with current annual production of 4.9 million tonnes.Materials and methodsA consensus map of lentil (Lens culinaris ssp. culinaris Medikus) was constructed using three different lentils recombinant inbred line (RIL) populations, including “CDC Redberry” x “ILL7502” (LR8), “ILL8006” x “CDC Milestone” (LR11) and “PI320937” x “Eston” (LR39).ResultsThe lentil consensus map was composed of 9,793 DArT markers, covered a total of 977.47 cM with an average distance of 0.10 cM between adjacent markers and constructed 7 linkage groups representing 7 chromosomes of the lentil genome. The consensus map had no gap larger than 12.67 cM and only 5 gaps were found to be between 12.67 cM and 6.0 cM (on LG3 and LG4). The localization of the SNP markers on the lentil consensus map were in general consistent with their localization on the three individual genetic linkage maps and the lentil consensus map has longer map length, higher marker density and shorter average distance between the adjacent markers compared to the component linkage maps.ConclusionThis high-density consensus map could provide insight into the lentil genome. The consensus map could also help to construct a physical map using a Bacterial Artificial Chromosome library and map based cloning studies. Sequence information of DArT may help localization of orientation scaffolds from Next Generation Sequencing data.

Identification, development, and application of cross-species intron-spanning markers in lentil (Lens culinaris Medik.)

Lentil (Lens culinaris Medik) is one of the most important food legumes in the world. The use in lentil of molecular marker-assisted breeding is limited, owing to the low availability of polymorphic markers. In the present study, we developed a set of polymorphic intron-spanning markers (ISMs) using a cross-species mapping approach. In this approach, putative unique transcripts (PUTs) of L. culinaris were mapped onto the Medicago truncatula genome, exploiting its closeness with the lentil genome. Spliced alignment of the PUTs resulted in a total of 25,717 alignments, allowing the development of 1703 ISMs. From these, a subset of 105 ISMs were synthesized and validated with a 51% amplification success rate in 32 lentil genotypes. Of these ISMs, 40 (74%) were polymorphic and generated 2–11 alleles per locus in a genetically diverse panel of 32 lentil genotypes including wild species. This set of polymorphic ISMs along with their functional annotation data will be useful in lentil breeding.

Marker-Trait Association Analysis of Iron and Zinc Concentration in Lentil (Lens culinaris Medik.) Seeds.

Lentil ( Medik.) seeds are relatively rich in iron (Fe) and zinc (Zn), making lentil a potential crop to aid in the global battle against human micronutrient deficiency. Understanding the genetic basis for uptake of seed Fe and Zn is required to increase sustainable concentrations of these minerals in seeds. The objectives of this study were to characterize genetic variation in seed Fe and Zn concentration and to identify molecular markers associated with these traits across diverse lentil accessions. A set of 138 cultivated lentil accessions from 34 countries were evaluated in four environments (2 sites × 2 yr) in Saskatchewan, Canada. The collection was genotyped using 1150 single-nucleotide polymorphism (SNP) markers that are distributed across the lentil genome. The germplasm tested exhibited a wide range of variation for seed Fe and Zn concentration. The marker-trait association analysis detected two SNP markers tightly linked to seed Fe and one linked to seed Zn concentration (-log10 ≥ 4.36). Additional markers were detected at -log10 ≥ 3.06. A number of putative candidate genes underlying detected loci encode Fe- and Zn-related functions. This study provides insight into the genetics of seed Fe and Zn concentration of lentil and opportunities for marker-assisted selection to improve micronutrient concentration as part of micronutrient biofortification programs.

Analysis of an intraspecific RIL population uncovers genomic segments harbouring multiple QTL for seed relevant traits in lentil (Lens culinaris L.).

Improving seed related traits remains key objective in lentil breeding. In recent years, genomic resources have shown great promise to accelerate crop improvement. However, limited genomic resources in lentil greatly restrict the use of genomics assisted breeding. The present investigation aims to build an intraspecific genetic linkage map and identify the QTL associated with important seed relevant traits using 94 recombinant inbreds (WA 8649090×Precoz). A total of 288 polymorphic DNA markers including simple sequence repeat(SSR), inter simple sequence repeat (ISSR)and random amplified polymorphic DNA(RAPD)were assayed on mapping population. The resultant genetic linkage map comprised 220 loci spanning 604.2cM of the lentil genome, with average inter-marker distance of 2.74cM. QTL mapping in this RIL population uncovered a total of18 QTL encompassing nine major and nine minor QTL. All major QTL were detected for seed related traits viz., seed diameter(SD), seed thickness(ST), seed weight (SW) and seed plumpness (SP)across two locations. A considerable proportion of the phenotypic variation (PV) was accounted to these QTL. For instance, one major QTL on LG5 controlling SW (QTL 15) explained 50% PV in one location, while the same QTL accounted for 34.18% PV in other location. Importantly, the genomic region containing multiple QTL for different seed traits was mapped to a 17-cM region on LG5. The genomic region harbouring QTL for multiple traits opens up exciting opportunities for genomics assisted improvement of lentil.

Obtaining retrotransposon sequences, analysis of their genomic distribution and use of retrotransposon-derived genetic markers in lentil (Lens culinaris Medik.).

Retrotransposons with long terminal repeats (LTR-RTs) are widespread mobile elements in eukaryotic genomes. We obtained a total of 81 partial LTR-RT sequences from lentil corresponding to internal retrotransposon components and LTRs. Sequences were obtained by PCR from genomic DNA. Approximately 37% of the LTR-RT internal sequences presented premature stop codons, pointing out that these elements must be non-autonomous. LTR sequences were obtained using the iPBS technique which amplifies sequences between LTR-RTs. A total of 193 retrotransposon-derived genetic markers, mainly iPBS, were used to obtain a genetic linkage map from 94 F7 inbred recombinant lines derived from the cross between the cultivar Lupa and the wild ancestor L. culinaris subsp. orientalis. The genetic map included 136 markers located in eight linkage groups. Clusters of tightly linked retrotransposon-derived markers were detected in linkage groups LG1, LG2, and LG6, hence denoting a non-random genomic distribution. Phylogenetic analyses identified the LTR-RT families in which internal and LTR sequences are included. Ty3-gypsy elements were more frequent than Ty1-copia, mainly due to the high Ogre element frequency in lentil, as also occurs in other species of the tribe Vicieae. LTR and internal sequences were used to analyze in silico their distribution among the contigs of the lentil draft genome. Up to 8.8% of the lentil contigs evidenced the presence of at least one LTR-RT similar sequence. A statistical analysis suggested a non-random distribution of these elements within of the lentil genome. In most cases (between 97% and 72%, depending on the LTR-RT type) none of the internal sequences flanked by the LTR sequence pair was detected, suggesting that defective and non-autonomous LTR-RTs are very frequent in lentil. Results support that LTR-RTs are abundant and widespread throughout of the lentil genome and that they are a suitable source of genetic markers useful to carry out further genetic analyses.

QTLs for iron concentration in seeds of the cultivated lentil (Lens culinaris Medic.) via genotyping by sequencing

Lentil, Lens culinaris Medic., is an important cool season grain legume because of the high level of iron (Fe) in its seeds since Fe deficiency is widespread and causes anemia. Thus, identifying genes controlling Fe concentration in the seed was needed for mapping in the lentil genome. The objectives of this study were to (i) detect phenotypic variation in Fe concentration in the seeds of a recombinant inbred line (RIL) population, (ii) construct a high-density linkage map using genotyping by sequencing (GBS), and (iii) identify localization of the quantitative trait loci (QTLs) controlling genes for Fe concentration in lentil seeds. In this work, Fe concentration in seeds of the RIL population ranged from 37.2 to 175.7 mg per kg. A linkage map was constructed covering 497.1 cM with a total of 4177 SNP markers. A total of 21 QTL regions explaining 5.9%-14.0% of the phenotypic variation were identified on six linkage groups (LG1, 2, 4, 5, 6, and 7) with LOD scores ranging from 3.00 to 4.45. This is the first report on the construction of a high-density linkage map through GBS and mapping of QTLs controlling iron uptake in lentil. Identification of these genomic regions will be useful for future biofortification studies to develop new varieties of lentil with high Fe concentrations.

Generation and Characterisation of a Reference Transcriptome for Lentil (Lens culinaris Medik.).

RNA-Seq using second-generation sequencing technologies permits generation of a reference unigene set for a given species, in the absence of a well-annotated genome sequence, supporting functional genomics studies, gene characterisation and detailed expression analysis for specific morphophysiological or environmental stress response traits. A reference unigene set for lentil has been developed, consisting of 58,986 contigs and scaffolds with an N50 length of 1719 bp. Comparison to gene complements from related species, reference protein databases, previously published lentil transcriptomes and a draft genome sequence validated the current dataset in terms of degree of completeness and utility. A large proportion (98%) of unigenes were expressed in more than one tissue, at varying levels. Candidate genes associated with mechanisms of tolerance to both boron toxicity and time of flowering were identified, which can eventually be used for the development of gene-based markers. This study has provided a comprehensive, assembled and annotated reference gene set for lentil that can be used for multiple applications, permitting identification of genes for pathway-specific expression analysis, genetic modification approaches, development of resources for genotypic analysis, and assistance in the annotation of a future lentil genome sequence.

Improving the nutritional value of pulse crops

Improving the nutritional value of pulse crops

Genetic Diversity of Cultivated Lentil (Lens culinaris Medik.) and Its Relation to the World's Agro-ecological Zones.

Assessment of genetic diversity and population structure of germplasm collections plays a critical role in supporting conservation and crop genetic enhancement strategies. We used a cultivated lentil (Lens culinaris Medik.) collection consisting of 352 accessions originating from 54 diverse countries to estimate genetic diversity and genetic structure using 1194 polymorphic single nucleotide polymorphism (SNP) markers which span the lentil genome. Using principal coordinate analysis, population structure analysis and UPGMA cluster analysis, the accessions were categorized into three major groups that prominently reflected geographical origin (world's agro-ecological zones). The three clusters complemented the origins, pedigrees, and breeding histories of the germplasm. The three groups were (a) South Asia (sub-tropical savannah), (b) Mediterranean, and (c) northern temperate. Based on the results from this study, it is also clear that breeding programs still have considerable genetic diversity to mine within the cultivated lentil, as surveyed South Asian and Canadian germplasm revealed narrow genetic diversity.