Actinomycetes were isolated from sugarcane soils by improved Gao’s No.1 synthetic medium. Physiological and biochemical characteristics, molecular identification and phylogenetic analysis were carried out on 29 isolates selected by morphology. Antimicrobial activity of the soil actinomycetes to plant pathogenic fungi was studied using the Dual-culture method. The results of identification showed that only one strain was identified as Cellulomonas, while the other twenty-eight strain were identified as Streptomyces. The strain G27 was active against Cochliobolus sativus, Alternaria alternata and Magnaporthe oryzae. Their inhibition rates of mycelium growth were 82.66%, 67.85%, 65.88%, respectively. Sugarcane community structure of soil actinomycetes is simple, with Streptomyces showing dominance in sugarcane populations. G27 showed a development potential for the biological control of crop disease.