[Objective] The antifungal bacteria of plant wilt disease was screened and identified to provide foundation for the study on bio-control preparation of plant wilt disease.[Method] Confrontation culture method was adopted to screen the bio-control bacteria with good antifungal effect against plant wilt disease,Biolog bacteria automatic identification system and 16S rDNA sequence analysis method were selected to identify its taxonomic status,the biological safety of the strain towards cotton and mice was also determined.[Result] 12 bacterial strains were isolated from rhizosphere of cotton.Among those strains,5 isolates showed antifungal activity against F.decemcellulare Brick,F.oxysporum f.sp.Diathi,F.oxysporum f.sp.vasinfectum.The antifungal effect of KL-1 strain against three target strains of pathogen reached 69.09%,80.78% and 78.89% respectively.Identification results of Biolog bacteria automatic identification system and 16S rDNA sequence analysis method showed that KL-1strain was Bacillus amyloliquefaciens;primary determination results of biological safety also showed that the strain KL-1 was safe and non-toxic towards cotton and mice.[Conclusion] KL-1strain of B.amyloliquefaciens had antifungal effect against several pathogens of plant wilt diseases,which was safe and non-toxic towards cotton and mice,being the bio-control strain with research and development potential.