Development of primers for PCR diagnosis of Xiphinema species associated with Japanese traditional ornamental trees

摘要

Dagger nematodes (Xiphinema spp.) were detected from nine tree species in a survey conducted in three Japanese prefectures, Chiba, Saitama and Fukuoka, producing Japanese traditional ornamental trees. For PCR diagnosis of these nematodes, we designedsix species-specific primer pairs on the basis of a known sequence of COI region of mtDNA and a known sequence of 18S small subunit to ITS1 region of rDNA. The specificity of the six designed primer pairs was confirmed from the fact that only target species or a related species was PCR amplified. As results of species-specific PCR and a BLAST search of sequences of D2-D3 expansion segments of 28S rDNA, surveyed nematodes were estimated to be X. brevicolle, X. insigne,X. parachambersi ovX. hunaniense. Additionally, a primer set of multiplex PCR for comprehensive amplification of these four species and two other species (.Xiphinema sp., X bakeri) was developed.