Cotton stalk bark fibers (CSBF) were extracted by steam flash explosion, completed within 0.09 s, and the extracted fibers were compared with those obtained by conventional alkaline treatment. Results indicate that the optimum steam pressure was 2.5 MPa when steaming time was set to 2 min for extracting CSBF. Under the optimized conditions, the obtained CSBF had a cellulose content of 72%, length of 48 mm, fineness of 45 dtex, crystallinity index of 68, moisture regain of 8%, water retention of 98%, and tensile strength of 2.4 cN/dtex, which were similar to results obtained by conventional alkaline treatment. Compared with bark of cotton stalks, CSBF had lower moisture regain and water retention, and higher onset decomposition temperature. The results show that moderate steam flash explosion is a chemical-free, quick, and effective method for exploring the industrial applications of bark of cotton stalks as natural cellulose fibers.