Cross-Species PCR Amplification Performance of Wheat Stripe Rust Resistance Gene-Specific Primers in Common Bean (Phaseolus Vulgaris L.)

Abstract

PCR-based gene-specific primers are essential in crop genetics, but their cross-genome effectiveness is unclear. This study tested whether wheat (Triticum aestivum L.) stripe rust resistance markers Yr15 and Yr30 can be amplified in common bean (Phaseolus vulgaris L.). Genomic DNA from 30 bean genotypes was amplified by PCR, with wheat DNA and nuclease-free water serving as positive and negative controls. The PCR protocol used an initial denaturation at 94°C for 5 minutes, 40 cycles (94°C for 60 seconds, 55.8°C for Yr15 or 52.7°C for Yr30 for 60 seconds, 72°C for 45 seconds), and a final extension at 72°C for 10 minutes. PCR products were separated via agarose gel and visualized. Yr15 was not detected in any bean genotype, while Yr30 showed the expected 120 bp band in some genotypes with additional non-specific bands. These findings highlight that gene-specific primers may fail or yield ambiguous results in unrelated species, emphasizing the need for rigorous marker validation across species.