Pardosa pseudoannulata (Boesenberg et Strand, l906) is one of the most common wandering spiders in agricultural fields, and a potential bioindicator for heavy metal contamination. In this study, a high-throughput method was used for discovering the simple sequence repeat (SSR) genetic markers from this spider. In total, 2805 sequences containing a total of 3450 SSRs were identified from the previously constructed transcriptome database, with 516 of the sequences containing more than one SSR. The most abundant SSRs in P. pseudoannulata are di-nucleotide (42.32%) SSRs, followed by mono-nucleotide (41.83%). A/T (41.36%) repeats were the most abundant, followed by AT/AT (28.32%). A total of 6561 pairs of SSR primers were designed using Primer 5.0 software, and a subset of 26 SSR primer pairs was randomly selected and validated by RT-PCR. The SSR markers identified here will be an important resource for measuring genetic diversity and biological monitoring research in P. pseudoannulata.