BACKGROUND: pABA biosynthesis is a potential target for antifolate drugs. RESULTS: Rubreserine inhibits GAT-ADCS, an enzyme involved in pABA biosynthesis, and decreased the folate content in Arabidopsis and Toxoplasma. CONCLUSION: Specific inhibition of pABA synthesis induces growth limitation of plants and apicomplexan parasites. SIGNIFICANCE: GAT-ADCS is a valuable target in eukaryotes, and rubreserine is a novel scaffold for anti-parasitic drugs.