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In the past decades, the taxonomic status of the cyanobacterial family Phormidiaceae has always been chaotic and problematic. In this study, filamentous cyanobacteria were investigated in the east of China, and twenty strains isolated from different locations of Zhejiang Province were characterized. Using the polyphasic approach combining morphological, molecular and phylogenetic features, these strains were grouped within the members of the genera <i>Ancylothrix</i> and <i>Potamolinea</i>, the newly recorded genera of cyanobacteria in China. Based on the collected taxonomic information of the family Phormidiaceae, cyanobacterial systematics at family level were further discussed. This study provided a simple and efficient example to perform the phylogenetic evaluation for the monophyly and rationality of currently used families of cyanobacteria by using the regional strains based on the polyphasic approach.