In order to understand the potential functions of actin in the growth and development of Charophytes, bioinformatics methods were used to analyze the actin gene family in Chara braunii. A total of 16 actin genes were identified in Chara braunii, and they were renamed as CbACT1~CbACT16 according to their order on scaffolds with 361 to 1182 amino acids, and the relative molecular masses ranged form 39 886.71 to 117 256.72 Da, and protein isoelectric point was range from 4.68 to 8.93. Subcellular localization prediction indicated that 15 actin genes were located in the cytoplasm; and only one in the chloroplast. The secondary structure of the actin gene was mainly composed of random coils and α-helices. Only one pair of paralogous genes derived from segmental duplication was found in the collinearity analysis. The phylogenetic results divided actin genes into two subfamilies with the same subfamily tend to have similar exon distribution and more common motifs. Promoter cis-element analysis indicated that 16 actin genes were involved in many biological activities including biotic and abiotic stresses, hormone regulation and light response. Further tissue expression analysis suggested that 16 genes were differentially expressed in four different tissues, indicating that they have different functions in the growth and development of different tissues. Therefore, the actin gene family may be involved in the development of different tissues and response to biotic and abiotic stresses of Charophytes.