Detection of plant parasitic nematodes, Meloidogyne incognita and Pratylenchus coffeae by multiplex PCR using specific primers

Abstract We investigated the internal transcribed spacer (ITS) region of plant parasitic nematodes, namely the root-knot nematode, Meloidogyne incognita and root-lesion nematode, Pratylenchus coffeae that hamper the development of sustainable cropping systems in the southern region of Kyushu, Japan. In the current study, we attempted to detect M. incognita and P. coffeae using multiplex PCR using specific primer sets. M. incognita specific primer set (Mi1-Mi2) was designed based on the sequence of the ITS region of M. incognita. P. coffeae specific primer set (Pc1-Pc2) was designed according to both the report of Uehara et al. (Nematoiogica, 44: 357–368, 1998) and the sequence of P. coffeae obtained in this study. The use of multiplex PCR enable to detect two distinctive bands derived from the two kinds of plant parasitic nematodes. Furthermore, these two specific bands were detectable, when the genome of other nematodes was mixed in the template, with at least one nematode of the plant parasitic nematode...