Classification and Characterization of Species within the Genus Lens Using Genotyping-by-Sequencing (GBS)

TitleClassification and Characterization of Species within the Genus Lens Using Genotyping-by-Sequencing (GBS)
AuthorsWong Melissa M. L., Gujaria-Verma Neha, Ramsay Larissa, Yuan Hai Ying, Caron Carolyn, Diapari Marwan, Vandenberg Albert, Bett Kirstin E.
TypeJournal Article
Journal AbbreviationPLOS ONE
Publication Date2015 March 27


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.


Wong MML, Gujaria-Verma N, Ramsay L, Yuan HY, Caron C, et al. (2015) Classification and Characterization of Species within the Genus Lens Using Genotyping-by-Sequencing (GBS). PLoS ONE 10(3): e0122025. doi:10.1371/journal.pone.0122025

Related Species
Related Species 

Lentil (Lens culinaris Medik.) is an important pulse crop with annual production of 3-4 Mt across 70 countries (Cubero et al. 2009. DOI 10.1079/9781845934873.0000; pg. 13). Lentils are a good source of protein, carbohydrates, micronutrients and vitamins for human nutrition and is consumed in more than 120 countries. Furthermore, their small seed size and flat shape make them relatively quick cooking and easily decorticated compared to most other grain legumes (Sharpe et al. 2013. BMC Genomics. DOI 10.1186/1471-2164-14-192). The Lentil plant has a bushy growth habit with a height of about 40 cm; the seeds are lens-shaped and usually grow two per pod. ... [more]

Research Area
Research Area 
Breeding & Genetics

Plant breeding is the art and science of changing the traits of plants in order to produce desired characteristics. Plant breeding can be accomplished through many different techniques ranging from simply selecting plants with desirable characteristics for propagation, to more complex molecular techniques. ... [more]