Fusarium oxysporum causes Fusarium wilt syndrome in more than 120 different plant hosts, including globally important crops such as tomato, cotton, banana, melon, etc. F. oxysporum shows high host specificity in over 150 formae speciales and have been ranked in the top 10 plant fungal pathogens. Although three PMTs encoded by the pmt1, pmt2, and pmt4 are annotated in the genome of F. oxysporum, their functions have not been reported. As O-mannosylation is not found in plants, a comprehensive understanding of PMTs in F. oxysporum becomes attractive for the development of new strategy against Fusarium wilt. In order to understand the molecular mechanism of the differential functions of three PMTs, a comparative O-glycoproteome analysis of the pmt mutants were carried out.