Two populations of the genus Trophotylenchulus and 10 species of the genus Paratylenchus from Iran were characterized based on morphometric, morphological and molecular characters. Our observations on the two populations of Trophotylenchulus from Iran revealed that T. asoensis and T. okamotoi have been distinguished from T. arenarius, on the basis of the features which cannot be longer considered as stable diagnostic characters. One of the populations shows a mixed combination of the characters of T. arenarius and T. asoensis; it has morphometrics more similar to T. arenarius but shows affinities with T. asoensis in the tail terminus shape of females and second-stage juveniles (J2) and in having a reduced stylet in males. The other population fit well with T. okamotoi; it has females with generally bluntly rounded tails typical for T. okamotoi, but sometimes with finely rounded tail termini, like those of T. arenarius or T. asoensis. The sequences of D2–D3 expansion segments of 28 S rRNA gene for the two populations are identical with each other, but only 4 bp (0.67%) difference with T. arenarius sequence deposited in the GenBank. Considering no stable difference allow separating species, synonymy of T. asoensis and T. okamotoi with T. arenarius, which has already been proposed, is supported and confirmed here. All studied Paratylenchus species with stylets longer than 40 µm, except P. straeleni, formed a basal cluster to Cacopaurus pestis and species of Paratylenchus bearing stylets shorter than 40 µm; thus, validity of Gracilacus cannot be rejected using our data sets. However, the synonymy of Paratylenchoides was supported by the positioning of P. sheri within representatives of Paratylenchus in the inferred phylogenetic tree.
