Tropical ageratum (Ageratum conyzoides L.) is a problematic weed frequently observed in association with commercially important crops in Australian agroecosystems. Knowledge of the germination response of A. conyzoides is crucial for proactively managing this weed species, especially when herbicide resistance is involved. Herbicide screening and metsulfuron dose–response experiments were conducted on two separate populations of A. conyzoides (referred to as Sugarcane and Roadside) in an open environment to identify a metsulfuron-resistant population. Based on the survival percentage in the metsulfuron dose–response experiment, the Sugarcane population was found to be 54 times more resistant compared with the metsulfuron-susceptible population (Roadside). Subsequent laboratory experiments were performed to investigate the differential germination response of the two populations. No germination or emergence difference was observed between the Sugarcane and Roadside populations under various thermal regimes (15/5 to 35/25 C with a 12/12-h photoperiod), salinity levels (0 to 320 mM), osmotic potentials (0 to −1.6 MPa), and burial depths (1 to 4 cm). However, different environmental conditions significantly impacted the germination and emergence of A. conyzoides. Ageratum conyzoides germinated over a wide range of temperatures, with the highest germination rate (>90%) occurring at 30/20 C. With increasing levels of salinity, osmotic potential, and burial depth, the germination/emergence of A. conyzoides declined and was completely inhibited at 300 mM salinity, −0.8 MPa osmotic potential, and a 1-cm burial depth. The data generated from this study will be useful in developing a model-based approach to predict the occurrence of this weed species and thus aid in designing ecologically sustainable integrated weed management protocols.