Biological assays are widely used to monitor toxic and allelopathic substances. The present study aimed to evaluate the allelopathic, genotoxic, and antiproliferative potential of aqueous extracts of Psychotria brachypoda (Müll. Arg.) Britton and Psychotria birotula Smith & Downs in two concentrations on the germination and cell division of Eruca sativa Hill. seeds. The biological assay was conducted in a controlled growth chamber. For monitoring the allelopathic effect, the following variables were evaluated: total number of germinated seeds, seedling root length, germination velocity index, and germination percentage. The means were compared using the Tukey test and orthogonal contrasts were undertaken to better compare the variables. To evaluate the antiproliferative and genotoxic effects, seedling roots were collected and the squashing technique was followed for preparation of slides. The results of the present study demonstrated that the medicinal species Psychotria brachypoda and Psychotria birotula inhibited root growth, germination velocity index, and germination percentage in seeds of arugula, in addition to inhibiting cell division and inducing chromosomal alterations in Eruca sativa. We conclude that the studied species have alellopathic, genotoxic, and antiproliferative effects on Eruca sativa in both concentrations studied.