Nassella trichotoma (Nees) Hack. ex Arechav. (Serrated tussock) is an aggressive globally significant weed to agricultural and natural ecosystems. Herbicide resistant populations of this C3 perennial weed have emerged, increasing the need for effective wide-scale cultural control strategies. A thorough seed ecology study on two spatially distinct populations of N. trichotoma was conducted on this weed to identify differences in important environmental factors (drought, salinity, alternating temperature, photoperiod, burial depth, soil pH, artificial seed aging, and radiant heat) which influence seed dormancy. Seeds were collected from two spatially distinct populations; Gnarwarre (38 O 9' 8.892'' S, 144 O 7' 38.784'' E) and Ingliston (37O 40' 4.44'' S, 144 O 18' 39.24'' E) in December 2016 and February 2017, respectively. Twenty sterilized seeds were placed into Petri dishes lined with a single Whatman® No. 10 filter paper dampened with the relevant treatments solution and then incubated under the identified optimal alternating temperature and photoperiod regime of 25°C/15°C (light/dark, 12h/12h). For the burial depth treatment, 20 seeds were placed into plastic containers (10cm in diameter and 6cm in depth) and buried to the relevant depth in sterilized soil. All trials were monitored for 30 days and germination was indicated by 5mm exposure of the radicle and emergence was indicated by the exposure of the cotyledon. Each treatment had three replicates for each population, and each treatment was repeated to give a total of six replicates per treatment, per population. Nassella trichotoma was identified to be non-photoblastic, with germination (%) being similar under alternating light and dark and complete darkness conditions. With an increase of osmotic potential and salinity, a significant decline in germination was observed. There was no effect of pH on germination. Exposure to a radiant heat of 120°C for 9 minutes resulted in the lowest germination in the Ingliston population (33%) and the Gnarwarre population (60%). In the burial depth treatment, the Ingliston population and the Gnarwarre population had highest emergence of 75% and 80%, respectively at a depth of 1cm. Variation between the two populations was observed for the burial depth treatments; Gnarwarre had greater emergence than Ingliston from the 4cm burial depth, while Ingliston had greater emergence at the soil surface than Gnarwarre. The Gnarwarre population had greater overall germination than Ingliston, which could be attributed to the greater seed mass (0.86mg compared to 0.76mg, respectively). This study identifies that spatial variations in N. trichotoma's seed ecology are present between spatially distinct populations.