The neotropical rust fungus Prospodium tuberculatum was released in Australia in 2001 for the classical biological control of the invasive alien weed Lantana camara, following a risk assessment that included studies on its life-cycle, infection parameters and host range. This paper reports on the first two research areas. This rust is recorded as autoecious and microcyclic (urediniospores and teliospores only) in the literature, but the role of the teliospores had not been demonstrated. Extensive field surveys reported here found the rust to cycle solely through urediniospores on L. camara throughout most of its native range. However, at the high altitude extremes of its range, the rust has retained its ability to over-winter as teliospores. Attempts to clarify the function of the basidiospores and the conditions necessary to induce teliospore formation were unsuccessful under controlled conditions. A hypothesised connection between P. tuberculatum and Aecidium lantanae was rejected after DNA analysis. Investigations on spore storage showed that teliospores and urediniospores can survive for extended periods in liquid nitrogen. Glasshouse-based studies demonstrated that the urediniospores have wide temperature tolerance (10–25°C) for infection of L. camara, with an optimum near to 20°C. The minimum dew period requirement for plant infection was 9 h with the optimum close to 15 h.