Taiwan Agricultural Research Institute Taichung, Taichung, Taiwan (Republic of China)
Abstract Text:
Bacterial wilt is a significant disease affecting crops in Taiwan, caused by the bacteria Ralstonia pseudosolanacearum and R. solanacearum. To manage the disease promptly, a convenient detection method has been developed for use in front-line units. This method utilizes a specific primer pair, Rs F6/R8, which can be employed in polymerase chain reaction (PCR) and recombinase polymerase amplification (RPA) methods to detect both bacterial pathogens. In PCR tests, a specific 157 bp DNA fragment was successfully amplified from both pathogens, while no amplification occurred with negative control bacterial strains. To further enhance convenience, the primers were also applied in RPA method, using commercial DNA detection strips to identify the RPA products. The results indicated that the DNA detection strips effectively detected the RPA products from both pathogens, whereas no RPA products were found in the negative control bacterial strains. This RPA method has been applied to diagnose bacterial wilt disease in various crops, including tomato, eggplant, ginger, pumpkin, and bitter gourd. Positive signals in the RPA products were detected through DNA detection strips in infected crops, while negative signals were observed in healthy crops. This demonstrates that the RPA method developed in this study has the specificity to diagnose bacterial wilt in crops. The combination of RPA method and DNA detection strips is user-friendly, does not require expensive equipment, and provides rapid results, making it suitable for front-line disease diagnosis units.
