In this research, it was first established that Tridax, a global tropical weed species, evolved glyphosate resistance in the Ord River irrigation area in north-western Western Australia. This is the first report of glyphosate resistance in Tridax. The mechanism of glyphosate resistance was studied. Various possible resistance mechanisms were NOT responsible for resistance (EPSPS gene amplification, different glyphosate uptake or translocation). In this glyphosate-resistant Tridax population, the glyphosate resistance mechanism is a mutation in the EPSPS gene causing substitution at amino acid 102 (Thr-102-Ser).