Wild lettuce (Lactuca serriola L.) is a significant emerging agricultural and environmental weed in many countries. This invasive species is now naturalised in Australia and is claimed to cause significant losses within the agricultural industry. Sustainable management of wild lettuce has been hampered by a lack of detailed knowledge of its seed ecology. Laboratory-based studies were performed to examine the potential influence of environmental factors including temperature and light conditions, salinity, pH, moisture availability and burial depth on the germination and emergence of two spatially distant populations of wild lettuce. Results suggested that the germination of wild lettuce seeds occurred across a broad range of temperature conditions (12-h cycle: 30°C/20°C, 25°C/15°C and 17°C/7°C) for both populations. We also found that these seeds are non-photoblastic; germination was not affected by darkness, with >80% germination in darkness for both populations at all tested temperature ranges. Germination significantly declined as salinity and osmotic stress increased for both populations, with seeds from the Tempy population were more affected by NaCl >100 mM than seeds from Werribee, but in neither population was there any observed effect of pH on germination (>80% germination in both populations at all tested pH ranges). For both populations, germination significantly decreased as burial depth increased; however, the two populations differed with regard to response to burial depth treatment, whereby seeds from the Tempy population had higher emergence than those from Werribee at 0.5 cm burial depth. These results suggest that light-reducing management techniques such as mulching or use of crop residues will be unsuccessful for preventing germination of wild lettuce. By contrast, burial of seeds at a depth of at least 4 cm will significantly reduce their emergence.