Molecular screening of candidate defense-related genes involved in resistance to Verticillium wilt in luffa (Luffa aegyptiaca)

In the current research, primer sets were designed to amplify four defense-related genes (DRGs), including Ve, PRP۱, RPS۲, and chi, which are associated with defense responses against Verticillium dahliae in luffa (Luffa aegyptiaca), or sponge gourd. Four luffa genotypes, including, Angled, Northern Large, Northern White Seed, and Touri, were evaluated as potential sources of genetic resistance. DNA was extracted from the genotypes, and degenerate primers were designed based on conserved regions of the DRGs using the Primer۳ software. The designed primers exhibited high specificity, enabling successful amplification of target gene fragments. The specificity of these primers was validated via PCR, agarose gel electrophoresis, and amplicon sequencing. BLAST of the newly obtained sequences in GenBank showed high similarity to the receptor-like protein, pathogenesis-related protein type ۱ (PRP۱), resistance to Pseudomonas syringae ۲ (RPS۲) family, and chitinase (chi) family, which are encoded by the Ve, PRP۱, RPS۲, and chi genes, respectively. The Ve and PRP۱ homologs were found in all genotypes. However, the RPS۲ homologs were not observed in Angled and Touri, and the chi homolog was not detected in the Touri genotype. Inoculation of seedlings with V. dahliae revealed mild to severe symptoms in genotypes.

کلیدواژه ها:

Cucurbitaceae ، Degenerate primers ، DRGs ، Soil-borne pathogen