The Grassland Earless Dragon (Tympanocryptis spp.) species complex occurred in the temperate native grasslands of south-eastern Australia (main map), grouped within a single species (Tympanocryptis pinguicolla), which was listed as Endangered on the IUCN Red List. However, available genetic and genomic data provided strong evidence that a full taxonomic revision was warranted [10,22]. As a result, T. pinguicolla is now thought to have been restricted to the Melbourne region and may represent the first extinction of a reptile on mainland Australia. Two new species, Tympanocryptis osbornei and Tympanocryptis lineata, were described and have a high probability of a threatened status. Full details of this case study and an example ROI assessment are provided in the supplementary materials (S1 Text, S3 Fig, S3 Table). Image credits: T. pinguicolla (F. and C. Collet); T. osbornei and T. lineata (S. Wilson). Map layer: Bioregional Assessment Source Dataset (https://data.gov.au/data/dataset/0cb242e2-daed-4507-a42e-73892c0941a1). Vegetation layer: pre-1750 tussock grasslands Department of Environment and Energy. 2018. National Vegetation Information System (NVIS) Version 5.1—AUSTRALIA (https://www.environment.gov.au/land/native-vegetation/national-vegetation-information-system/data-products).