Abstract: Tachia guianensis (Gentianaceae), a Neotropical understory myrmecophyte, shelters ant colonies in its hollow trunks and branches (domatia). In turn, it is protected from defoliators and obtains nutrients from ant-produced wastes (myrmecotrophy). Aiming to verify if seasonality influences nitrogen assimilation via ant wastes using the stable isotope nitrogen-15, we first studied Tachia’s phenology and its seasonal leaf production, and then the life cycle of its two more frequent guest ant species. We found that leaf production was much higher during the rainy than the dry season. Mature guest ant colonies produced sexuals regardless of the season and the net weight of the waste piles inside the domatia did not vary between seasons, so that the availability of nutrients to their host plant is steady year-long. By providing the two most frequent mutualistic guest ant species with food enriched with nitrogen-15, we showed that Tachia individuals assimilate more nitrogen from ant wastes during the rainy season, when the plant is physiologically active, compared to the dry season. Thus, one can deduce that the increase in nitrogen assimilation during the rainy season is determined by the increase in Tachia’s physiological activity during that season. Information gathered through a bibliographic compilation confirms that none of the 15 ant species known to be associated with myrmecophytes for which the life cycle was studied is characterized by seasonal reproduction (which would result in fluctuating waste production). The same is true for 49.7% of 167 tropical ant species (seasonal production for the remaining species). We concluded that, in contrast to the non-seasonal ant colony reproductive cycle, Tachia’s phenology determines the myrmecotrophic assimilation rate. © 2020, Springer Nature Switzerland AG.