Vink, J. P., Soltani, N., Robinson, D. E., Tardif, F. J., Lawton, M. B. and Sikkema, P. H. 2012. Occurrence and distribution of glyphosate-resistant giant ragweed (Ambrosia trifidaL.) in southwestern Ontario. Can. J. Plant Sci. 92: 533–539. Giant ragweed is the first confirmed glyphosate-resistant (GR) weed in Canada. A survey was conducted to document the distribution of GR giant ragweed in southwestern Ontario. Giant ragweed seed was collected from 102 sites in Essex (70), Chatham-Kent (21), Lambton (10) and Waterloo counties (1) during the autumn of 2009 and 2010 prior to soybean harvest. Plants were grown in a growth room, sprayed with glyphosate at 1800 g a.e. ha^-1 and classified as resistant or susceptible. GR giant ragweed has been confirmed at 47 new locations in three counties (Essex, Chatham-Kent and Lambton). The results from this survey indicate that GR giant ragweed biotypes occur across a greater area in southwestern Ontario than originally thought. This survey provides an important baseline for future surveys in the province.