Isolation of TIR and nonTIR NBS-LRR resistance gene analogues and identification of molecular markers linked to a powdery mildew resistance locus in chestnut rose (Rosa roxburghii Tratt)

摘要: Toll and interleukin-1 receptor (TIR) nonTIR nucleotide binding site–leucine rich repeat (NBS–LRR) resistance gene analogues (RGAs) were obtained from chestnut rose (Rosa roxburghii Tratt) by two PCR-based amplification strategies (direct overlap extension amplification) with degenerate primers designed to the conserved P-loop, kinase-2, Gly-Leu-Pro-Leu (GLPL) motifs within NBS domain of plant (R gene) products. Thirty-four 65 cloned PCR fragments contained a continuous open reading frame (ORF) their predicted protein products showed homology NBS–LRR class R proteins in GenBank database. These 34 sequences exhibited wide range (19.5–99.4%) sequence identity among them classified into distinct groups phylogenetic analysis. The first group consisted 23 seemed belong RGAs, since they specific (RNBS-A-nonTIR motif) that are often present coiled-coil genes. second comprised 11 found TIR Restriction fragment length polymorphic (RFLP) markers developed some RGAs used for mapping powdery mildew genes rose. Three markers, RGA22C, RGA4A, RGA7B, identified be linked locus, designated CRPM1 1, which accounted 72% variation phenotype an F1 segregating population. To our knowledge, this is report on isolation, analysis potential utilization as genetic