A direct 15‐second DNA extraction method from pinewood strips facilitates field surveillance and quarantine of Bursaphelenchus xylophilus (pinewood nematode)

BACKGROUND: Pine wilt disease, caused by the pinewood nematode (Bursaphelenchus xylophilus), poses a significant threat to pine trees. Therefore rapid and accurate molecular detection of B. xylophilus is particularly important for the prevention and control of pine wilt disease; however, traditional nematode DNA extraction methods are complex, time-consuming and not suitable for on-site sample collection. Hence, to overcome this limitation, we developed a practical applied diagnostic technology based on direct DNA extraction from pinewood strips. RESULTS: Through systematic optimization, the handheld ultrasonic disruptor was identified as the most effective tool for cell lysis, and increment borer sampling from the inner rings of infected pine trees yielded the highest detection accuracy. We confirmed that our method can extract DNA from B. xylophilus within 15 s using pinewood strips and that the obtained DNA can be directly used for polymerase chain reaction (PCR) amplification. Sensitivity tests confirmed that the method could detect a single nematode in spiked, artificially inoculated pinewood strips, while validation with 30 field-collected pinewood samples achieved 100% consistency with traditional morphological identification. CONCLUSION: This 15-s DNA extraction technique facilitates faster, simpler, and more cost-effective diagnosis of pine wilt disease, offering a valuable tool for future disease management and control.