Abstract:A total of 14 501 unigenes in Fusarium oxysporum f. sp. lycopersic were screened by using bioinformatics software to search for Simple sequence repeat (SSR) motifs, 466 SSRs were sought out, distributing in 425 unigenes and the occurrence freguency was 2. 93%. Trinucleotide repeats were major types among the obtained unigenes, accounting for 65. 88%. AGC/CTG (16. 52%) and AAG/CTT (12. 66%) were the most abundant motifs. Based on the flanking sequences, 451 primer pairs were designed by using Premier 3 software. These results showed that it is an effective and feasible approach to developing SSR markers based on genome database in F. oxysporum f. sp. lycopersic, and that there were abundant SSR loci in the genome of F. oxysporum f. sp. lycopersic.