| description |
Resistance gene enrichment and sequencing (RenSeq) is a method that enables de novo assembly and (re)annotation of the NB-LRR gene complement in plant species. We refined RenSeq to capture and sequence 3.2kb fragments (average NB-LRR gene length) with long read sequencing offered by the PacBio RSII. We applied it to a previously un-sequenced S. americanum accession what allowed de novo assembly of full NB-LRR repertoires. Assembly of long flanking regions enabled analysis of complex NB-LRR loci carrying multiple NB-LRR genes and revealed domain fusions and NB-LRR-pairs. Most importantly, combining long read assembly of the resistant parent, bulked segregant analysis and RenSeq cDNA with short read RenSeq data, we quickly cloned a novel broad-spectrum resistance gene against P. infestans. |