Resumo. Abstract: Aim This contribution reports the first regional occurrence of Pleurosira laevis in the Masuleh River, Iran and additionally describes the pattern of occurrence along the Masuleh River and among four substrate types. The aim of this study was to assess the effects of substrate type and physical and chemical variables on distribution of centric diatom P. laevis. Methods At each station, triplicate samples were collected from 4 substrata. Epilithic (assemblages on rock), epidendric (assemblages on wood), epipsammic (assemblages on sand), and epipelic (assemblages on mud) diatom and water quality sampling was done four times at 5 stations. Physical and chemical variables including total nitrate, total phosphate, silicate, Fe2+, EC, and pH were also determined. Samples preserved in 2% for formalin solution and transferred to the laboratory, boiled with acid (HNO3: H2SO4; 2:1), centrifuged, identified, and counted. Cluster analysis was performed to indicate the main differences and similarities in diatom abundance amongst substrates sampled and amongst sampling stations. Results The highest (12.54 ± 1.54) and lowest (0.74 ± 0.10) abundance was obtained from wooden and muddy substrates at stations S5 and S4, respectively. The highest abundance was observed in the wooden substrate among all of sampling stations where showed significant differences (P < 0.05) with the other substrates. The hierarchical clustering based on relative abundance of P. laevis distinguished two clusters amongst the four substrate types. Rougher and more stable substrates (wood, sand, and stone) clearly separated from smooth and unstable substrates (mud). Based on relative abundance of P. laevis on rougher substrates, epidendric and epipsammic species had the highest similarity. Results of Pearson correlation showed that relative abundance of P. laevis had a significant correlation with EC, TN, TP, and Fe2+ concentrations (P < 0.05) whilst no significant correlation was observed with pH, temperature, and SiO2 concentration (P > 0.05). Conclusion We conclude that wood substrata can be substituted for one another during field surveys. Results from this study demonstrate that the type and roughness of the substrate both influence the attachment of P. laevis and its subsequent growth conditions.