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WRKY transcription factors play central roles in developmental processes and stress responses of wheat. Most WRKY proteins of the same group (Group III) have a similar function in abiotic stress responses in plants. <i>TaWRKY46</i>, a member of Group III, was up-regulated by PEG treatment. TaWRKY46-GFP fusion proteins localize to the nucleus in wheat mesophyll protoplasts. Overexpression of <i>TaWRKY46</i> enhanced osmotic stress tolerance in transgenic <i>Arabidopsis thaliana</i> plants, which was mainly demonstrated by transgenic <i>Arabidopsis</i> plants forming higher germination rate and longer root length on 1/2 Murashige and Skoog (MS) medium containing mannitol. Furthermore, the expression of several stress-related genes (<i>P5CS1</i>, <i>RD29B, DREB2A, ABF3, CBF2,</i> and <i>CBF3</i>) was significantly increased in <i>TaWRKY46-</i>overexpressing transgenic <i>Arabidopsis</i> plants after mannitol treatment. Taken together, these findings proposed that TaWRKY46 possesses vital functions in improving drought tolerance through ABA-dependent and ABA-independent pathways when plants are exposed to adverse osmotic conditions. <i>TaWRKY46</i> can be taken as a candidate gene for transgenic breeding against osmotic stress in wheat. It can further complement and improve the information of the WRKY family members of Group III.