Allele Mining in Solanum Germplasm: Cloning and Characterization of RB-Homologous Gene Fragments from Late Blight Resistant Wild Potato Species

Abstract

The late blight disease can be managed by introduction of resistance (R) genes from the wild Solanum species into the cultivated potato. The R genes are mostly comprised of the nucleotide binding site-leucine rich repeat (NBS-LRR) domains and share nucleotide sequence homology in the crop species. In this study, we used potato R gene-specific primers to amplify homologous genes from wild species. A total of 39 wild species were tested for late blight resistance by challenge inoculation of Phytophthora infestans under controlled conditions. Of these, only 15 species were highly resistant (HR) and these were PCR (polymerase chain reaction) amplified by 53 primers representing 21 R genes of potato. Further, only single, distinct, and reproducible gene fragments were cloned and sequenced. Following sequence processing and analysis, 17 non-redundant sequences of RB-homologous genes were identified with uninterrupted open reading frames (ORFs) and nucleotide sequence homologies to known late blight R genes. Finally, 17 RB-homologous gene fragments amplified by the primers of the RB gene were isolated from 11 wild species. The isolation and characterization of 17 RB-homologous gene fragments from wild potato species may serve as an important genomic resource for the novel gene discovery in late blight resistance breeding programs.