Molecular diagnosis of the Seville root-knot nematode Meloidogyne hispanica based on SCAR marker

Abstract Meloidogyne hispanica has received significant attention as it poses a major threat to various crops due to its expanding geographical distribution. Consequently, there is an urgent need for specific diagnostic techniques targeting M. hispanica . However, the development of effective molecular diagnostic methods has been hindered by the lack of genomic information. To address this challenge, we developed a species-specific SCAR-PCR assay based on a RAPD marker of M. hispanica in this study. The SCAR primers were designed from the RAPD amplicon, yielding a 385-bp fragment exclusively amplified from the genome of M. hispanica . This method successfully amplified DNA from M. hispanica at all developmental stages and from infected soil samples. It exhibited high sensitivity, with detection limits equivalent to 1/1280 of a single female and 1/160 of a second-stage juvenile. This method provides a reliable tool for the specific detection of M. hispanica , enabling timely monitoring and effective containment of its spread.