Ragweed parthenium is a highly invasive weed species in several countries, including Australia. Laboratory experiments were conducted to evaluate the effect of temperature, light, salinity, pH, and moisture on germination of two Australian biotypes of ragweed parthenium: Clermont (highly invasive) and Toogoolawah (noninvasive). Although seeds of both biotypes could germinate under complete darkness, germination was improved by 20% to 49% under a 12-h photoperiod. Both biotypes germinated over a wide range of constant (8 to 35 C), and alternating day/night (15/5 to 35/25 C) temperatures. However, the Clermont biotype exhibited significantly higher germination than Toogoolawah biotype over the range of temperatures studied. Highest germination of Clermont (100%) and Toogoolawah (97%) was observed at constant temperatures of 14 to 23 C and 23 C, respectively. The best alternating day/night temperature for germination of both biotypes was 25/15 C. Clermont also germinated better than Toogoolawah under osmotic- and salt-stress conditions. Osmotic stress had moderate negative effects on germination, with 52% and 36% of the Clermont and Toogoolawah seeds able to germinate at −0.60 MPa, respectively. Complete germination inhibition for both biotypes was observed at an osmotic potential of −1.2 MPa. Both biotypes also germinated at a very high sodium chloride (NaCl) concentration of 250 mM. A 50% reduction in germination of Toogoolawah and Clermont was caused by 99 and 154 mM NaCl, respectively. Germination of the Clermont biotype was not affected by a wide range of pH (4.0 to 10.0), whereas the strong acidic and alkaline pH levels (4.0 and 10.0) caused 18% and 25% reductions in germination of the Toogoolawah biotype compared with control. The Clermont biotype had a higher ability to germinate across all treatments compared with the Toogoolawah biotype, which might be a contributing factor toward the high invasive ability of the former compared with the latter.