Transcriptomics and QTL analysis to identifying candidate genes for yellow rust resistance in wheat

A winter wheat bi-parental population of 109 lines segregating for yellow rust resistance was sown in the field in Svalöv, Sweden in the autumn of 2013. Scoring was done at the booting stage (Zadoks stage 41-49) in the spring of 2014 with a scale of 1 (high resistant) to 9 (highly susceptible). Leaf material was collected from penultimate leaves pooled from three-four plants from each line of the segregating population at the booting stage (Zadoks stage 41-49). Leaves were flash frozen in liquid nitrogen and stored at -180oC until further processing. Total RNA was extracted with RNeasy Plant Mini kit (Qiagen) including DNase treatment (RNase-free DNase set, Qiagen). Concentration and quality of the RNA was estimated in Experion™ Automated Electrophoresis System (Bio-Rad Laboratories, Hercules, USA). CDNA library construction and sequencing of 20 samples with 9 resistant and 11 susceptible lines for yellow rust were done at the SciLifeLab (Stockholm, Sweden). Sequencing was performed on Illumina Hiseq 2500 instrument by paired-end sequencing of 150bp length as per manufacturer’s instructions.