The potato rot nematode (Ditylenchus destructor) has been recorded from several countries and causes considerable yield loss of potato. During 2018-2020, the potato rot nematode, D.destructor, Thorne, 1945 were extracted from potato tubers collected from six regions of Iran and the Russian Federation. These populations were described and compared for the first time based on their morphological, morphometric, and molecular characteristics. Furthermore, two new primer pairs were developed to exactly identify the nematode. The morphological and morphometric features were generally in agreement with those published for D. destructor. The rRNA gene of the nematodes was amplified and sequenced using two primer sets: dsn.1 R/F and dsn.2 R/F. New sequences were deposited in the GenBank under accession numbers MN122076, MN307126, MN307128, MN493767, MN658597, MN658599, MN658637, MN658638. The identification was further supported by PCR with designed species-specific primers for this species.