During 2014 and 2015, symptoms of root rot were observed in tobacco fields located in Veracruz, Mexico. Phytophthora colonies developing rosaceous pattern were consistently isolated on potato dextrose agar (PDA) and V8 juice agar media. Sporangia were papillate, noncaducous, measuring 40 to 49 x 33 to 37 μm, and with short pedicels. The sporangial shape was ovoid or ellipsoid. Chlamydospores were mostly solitary, intercalary, or terminal, measuring 29-33 μm in diameter. The oomycete was identified as Phytophthora nicotianae based on descriptions of Gallegly and Hong (2008). To confirm the identification, genomic DNA from a representative isolate was extracted, and the internal transcribed spacer (ITS) region was amplified by PCR and sequenced using the primers ITS5 and ITS4 (White et al., 1990). The resulting 896 bp sequence was deposited as GenBank Accession No. KR906059. BLASTn analysis showed a 97% similarity with those of P. nicotianae (GenBank Accession No. AB688370). Pathogenicity tests were performed twice on 20 healthy tobacco plants by immersion of roots in a zoospore suspension adjusted to 1 x 104 zoospores ml-1. Ten tobacco plants inoculated with sterile distilled water served as controls. Inoculated plants developed root rot symptoms after 10 days, whereas no symptoms were observed on the control plants. The oomycete present on inoculated plants was morphologically identical to that originally observed on diseased plants, thus fulfilling Koch’s postulates. To our knowledge, this is the first report of tobacco root rot caused by Phytophthora nicotianae in Mexico.