Molecular Characterization and Phylogenetic Analysis of Hoplolaimus stephanus.

Three Hoplolaimus stephanus populations were characterized morphologically, both by morphometrics and by SEM observations. These populations were used to develop a rapid and accurate molecular identification method for the species, which is useful because of the high level of morphological similarity between H. stephanus and H. galeatus. Species-specific primers for H. stephanus, amplifying a distinct fragment (260 bp) of the nuclear ribosomal internal transcribed spacer1 (ITS1), can be used in multiplex PCR along with previously developed primers for other common Hoplolaimus species. We also infer phylogenetic relationships among H. stephanus, the closely-related H. galeatus, and several other Hoplolaimus species, using sequences of the actin gene, ITS1 and LSUD.