Nowadays, plant allelopathy, as a new type of biological algal and/or bacterial inhibition technology, has attracted extensive attention. Eight substances were isolated and identified from Jussiaea stipulacea Ohwi, and five concentration gradients, as well as a control (0, 1.25, 5, 10, 20, and 50 mg/L) were set, with three parallels in each group, and then sampled and detected at 24, 48, 72, and 96 h. When the concentration was 50 mg/L, the inhibition rate of Anabaena was as high as 74.8%, 69.2%, and 70.7% for ursolic acid, kaempferol, and luteolin, respectively. Streptococcus iniae and Aeromonas hydrophila were cultured to a logarithmic phase, and their final concentrations reached 1000, 500, 250, 125, 62.50, 31.25, 15.63, and 7.81 μg/mL. Luteolin and gallic acid showed an inhibitory effect on S iniae and A hydrophila at 1000 μg/mL. We found that allelochemicals also had a certain bacteriostatic effect, among which luteolin has great development potential.