A new species of the genus Nothotylenchus, N. persicus n. sp. was collected around the roots of grapevine and is described and illustrated herein based on morphological and molecular studies. The new species is characterised by a body length of 776-900 μm, delicate stylet 5-6 μm long, six lines in the lateral field, post-vulval uterine sac short, 10-18 μm long, female tail elongate-conoid with pointed terminus, and bursa covering 40-45% of tail length. Morphologically, N. persicus n. sp. appears closer to four known species of the genus, namely: N. hexaglyphus, N. affinis, N. medians and N. taylori. The results of phylogenetic analyses based on sequences of D2-D3 expansion region of 28S rRNA gene confirmed the close molecular relationship between N. persicus n. sp. and other anguinids, but Nothotylenchus claded separately from Ditylenchus species.