In addition, germplasm collections that possess a full range of genetic diversity and phenotypic expressions have the potential to serve as platforms for association studies to identify statistically significant relationships between polymorphic markers and genes of economic and biological merit [34]. In the current study, we focused on distilling the molecular diversity
and genetic structure of 298 homozygous lettuce lines and using this information to assess genome-wide marker-trait associations between SNP markers and 10 horticultural traits. Three hundred and eighty-four individual plants sampled from 356 accessions were used selleck chemical in this study. For some accessions, more than one plant per accession was sampled based on observed differences in morphology. These accessions were collected worldwide during 1930s–2010s and are maintained at the USDA-ARS Selleck Seliciclib Western Regional Plant Introduction Station (WRPIS) in Pullman, Washington. Genomic DNA was extracted from single plants using the DNeasy 96 Plant
Kit (Qiagen, Valencia, CA, USA). Quality and quantity of extracted DNA samples were evaluated with Fluoroskan Ascent FL (Thermo Scientific, Hudson, NH, USA). The SNP genotyping assay was carried out at the UC Davis Genome Center using 250 ng of genomic DNA per sample and the LSGermOPA panel targeting 384 EST-derived SNP loci. A more detailed description of the genotyping procedure can be found in our previous study [30]. Seeds of the genotyped plants were harvested and planted in 2011 and 2012 at the WRPIS Central Ferry Research Farm, Central Ferry, WA, for confirming Thymidylate synthase homozygosity within accessions and for phenotypic evaluation. The phenotypic traits surveyed in the field from June to November, 2011 and 2012, included horticultural type, leaf color, bolting date, flowering date, leaf anthocyanin, stem anthocyanin, stem fasciation, leaf margin undulation, leaf blistering,
and seed coat color. Bolting and flowering dates were recorded when the plant rachis was 10 cm and the terminal flower of the main axis was fully open, respectively. Leaf color, anthocyanin, margin undulation and blistering and horticultural type were recorded before the bolting stage; stem anthocyanin, and fasciation were recorded after bolting. Seed coat color was observed after harvest. A cluster analysis was conducted using the UPGMA (unweighted pair group method with arithmetic mean) based on the allele-sharing distance by PowerMarker version 3.25 [35] and the resulting tree was displayed using the software Mega4 [36]. Population structure was assessed using the software package STRUCTURE 2.3.3 [37] that utilizes a Bayesian algorithm to assign accessions to putative populations (K). Inferred information about population structure and the degree of admixture can subsequently be used as a co-factor in association mapping.