Special Antarctic Blend (SAB) is a diesel fuel dominated by aliphatic hydrocarbons that is commonly used in Antarctic and subantarctic regions. The past and present use of SAB fuel at Australia's scientific research stations has resulted in multiple spills, contaminating soils in these pristine areas. Despite this, no soil quality guidelines or remediation targets have been developed for the region, primarily due to the lack of established indigenous test species and subsequent biological effects data. In this study, twelve plant species native to subantarctic regions were collected from Macquarie Island and evaluated to determine their suitably for use in laboratory-based toxicity testing, using germination success and seedling growth (shoot and root length) as endpoints. Two soil types (low and high organic carbon (OC)) were investigated to reflect the variable OC content found in soils on Macquarie Island. These soils were spiked with SAB fuel and aged for 14 days to generate a concentration series of SAB-contaminated soils. Exposure doses were quantified as the concentration of total petroleum hydrocarbons (TPH, nC9–nC18) on a soil dry mass basis. Seven species successfully germinated on control soils under laboratory conditions, and four of these species (Colobanthus muscoides Hook.f., Deschampsia chapmanii Petrie, Epilobium pendunculare A.Cunn. and Luzula crinita Hook.f.) showed a dose-dependent inhibition of germination when exposed to SAB-contaminated soils. Contaminated soils with low OC were generally more toxic to plants than high organic carbon soils. Increasing soil-TPH concentrations significantly inhibited shoot and root growth, and root length was identified as the most sensitive endpoint. Although the test species were tolerant to SAB-contaminated soils in germination assays, development of early life stages (up to 28 days) were generally more sensitive indicator of exposure effects, and may be more useful endpoints for future testing.