The Charales, commonly known as stoneworts, are a group of highly complex green algae that comprise one extant family (Characeae) with six genera. The aim of this study was to collect Charophytes in eastern and north of Iran and check the distribution maps of species in Iran. Samples were collected from 103 localities and 7 species have been determined with Chara braunii Gm., Chara globata Mig. and Lamprothamnium papulosum (Wallr.) J. Gr. recorded for the first time in Iran. Although Chara vulgaris and Chara gymnophylla were reported as widespread in north and eastern Iran, specimens of Chara vulgaris were collected in a wide variety of habitats from most of the provinces rendering this taxa the most abundant Charophytes in Iran (88 populations). Other species were restricted to few locations: Chara braunii, a cosmopolitan species, was limited to two locations in north Iran. Chara globata was found in a single location (Targhrud Lake). Most specimens of Chara contraria and Chara connivens were collected from South West Iran (260–350 m elevation). Lamprothamnium papulosum, a rather uncommon species in Asia, was found in a single lake with marine water characteristics in northeast Iran.