Two new amphimictic species, Rotylenchus iranicus n. sp. and R. conicaudatus n. sp., are described. Rotylenchus iranicus n. sp. is characterised mainly by an offset hemispherical lip region with 5-6, rarely 7, annuli, stylet 39-44 μm long, vulva located at 53-65% and rounded tail with 4-9 annuli and typical smooth tip. Morphologically this species is related to R. montanus, R. provincialis and R. aqualamus. Rotylenchus conicaudatus n. sp. is characterised mainly by a slightly offset conoid-rounded lip region with four or five, rarely six annuli, stylet 27-32 μm long, vulva located at 52-63% and conoid-rounded tail with 10-16 annuli and a typical annulated tip. Morphologically this species is related to R. pumilus, R. abnormecaudatus, R. acuspicaudatus and R. provincialis. The results of the phylogenetic analysis based on sequences of the D2-D3 expansion regions of 28S and ITS1-rRNA genes confirmed the species differentiation. Phylogenetic relationships with other species were difficult to assign using D2-D3 expansion regions of 28S. However, using ITS1-rRNA, R. conicaudatus n. sp. presented a close relationship with R. unisexus, whilst R. iranicus n. sp. was closely related to R. conicaudatus n. sp., R. unisexus, R. incultus and R. laurentinus.