Dirigent (DIR) genes play a pivotal role in plant development and stress adaptation. Manihot esculenta Crantz, commonly known as cassava, is a drought-resistant plant thriving in tropical and subtropical areas. It is extensively utilized for starch production, bioethanol, and animal feed. However, a comprehensive analysis of the DIR family genes remains unexplored in cassava, a crucial cash and forage crop in tropical and subtropical regions. In this study, we characterize a total of 26 cassava DIRs (MeDIRs) within the cassava genome, revealing their uneven distribution across 13 of the 18 chromosomes. Phylogenetic analysis classified these genes into four subfamilies: DIR-a, DIR-b/d, DIR-c, and DIR-e. Comparative synteny analysis with cassava and seven other plant species (Arabidopsis (Arabidopsis thaliana), poplar (Populus trichocarpa), soybean (Glycine max), tomato (Solanum lycopersicum), rice (Oryza sativa), maize (Zea mays), and wheat (Triticum aestivum)) provided insights into their likely evolution. We also predict protein interaction networks and identify cis-acting elements, elucidating the functional differences in MeDIR genes. Notably, MeDIR genes exhibited specific expression patterns across different tissues and in response to various abiotic and biotic stressors, such as pathogenic bacteria, cadmium chloride (CdCl₂), and atrazine. Further validation through quantitative real-time PCR (qRT-PCR) confirmed the response of DIR genes to osmotic and salt stress. These findings offer a comprehensive resource for understanding the characteristics and biological functions of MeDIR genes in cassava, enhancing our knowledge of plant stress adaptation mechanisms.