The pan-NLRome analysis based on 23 genomes reveals the diversity of NLRs in Brassica napus

Brassica napus, a globally significant oilseed crop, exhibits a wide distribution across diverse climatic zones. B. napus is being increasingly susceptible to distinct diseases, such as blackleg, clubroot and sclerotinia stem rot, leading to substantial reductions in yield. Nucleotide-binding site leucine-rich repeat genes (NLRs), the most pivotal family of resistance genes, can be effectively harnessed by identifying and uncovering their diversity to acquire premium disease-resistant gene resources. Here, we collected the genomes of 23 accessions and established the first comprehensive pan-NLRome in B. napus by leveraging multiple genomic resources. We observe significant variation in the number of NLR genes across different B. napus accessions, ranging from 189 to 474. Notably, TNL (TIR-NBS-LRR) genes constitute approximately half of the total count, indicating their predominant presence in B. napus. The number of NLRs in the C subgenome is significantly higher than that in the A subgenome, and chromosome C09 exhibits the highest density of NLR genes with featuring multiple NLR clusters. Domain analysis reveals that the integrated domains significantly enhance the diversity of NLRs, with B3 DNA binding, VQ, and zinc fingers being the most prevalent integrated domains. Pan-genomic analysis reveals that the core type of NLR genes, which is present in most accessions, constitutes approximately 58% of the total NLRs. Furthermore, we conduct a comparative analysis of the diversity of NLR genes across distinct ecotypes, leading to the identification of ecotype-specific NLRs and their integrated domains. In conclusion, our study effectively addresses the limitations of a single reference genome and provides valuable insights into the diversity of NLR genes in B. napus, thereby contributing to disease resistance breeding.