The invention discloses a Bacillus licheniformis 201, which is preserved in China Center for Type Culture Collection that is located in Wuhan University, Logosun, Wuchang, Wuhan, Hubei, and with CCTCC NO. of M207092. The invention simultaneously discloses a preparation method and applications of the Bacillus licheniformis 201, which has strong bacteriostasis of banana wilt fusarium, wide bacteriostasis spectrum and antagonism to various pathogenic fungi including banana wilt fusarium, watermelon wilt fusarium, tomato wilt fusarium, banana colletotrichum, gloeocystidium colletotrichum, corn leaf spot curvularia, cotton verticillium lecanii, gibberella zeae, and the like, and the Bacillus licheniformis 201 has high control efficiency, good environmental security and good development and application prospect.