Resistance gene enrichment and sequencing (RenSeq) of wild Solanum species

RenSeq is a NB-LRR gene-targeted, Resistance gene enrichment and sequencing method that enables discovery and annotation of pathogen resistance gene family members in plant genome sequences and operates independently of existing gene annotations. We successfully applied RenSeq to the sequenced potato Solanum tuberosum clone DM, and increased the number of identified NB-LRRs from 438 to 762. The majority of newly identified R gene loci reside in poorly- or previous un-annotated regions of the genome. We compared these NB-LRR genes and the corresponding oligonucleotide baits with the highest sequence similarity and demonstrated that ~80% sequence identity is sufficient for enrichment. We applied RenSeq to rapidly identify molecular markers that co-segregate with resistance towards the late blight pathogen Phytophthora infestans in two independent segregating populations involving the wild Solanum species S. berthaultii (Rpi-ber2) and S.ruiz-ceballosii (Rpi-rzc1).