Grain fructans play an important role in the physiology of wheat plants and also impact on the health of consumers of wheat-based products. Given the potential economic importance of fructan levels, if genetic variability could be identified for this trait, it may be a potentially useful breeding target for developing climate-resilient and nutritionally enhanced wheat varieties. The aim of the current study was to screen 78 genetically diverse Australian wheat varieties released between 1860 and 2015 to determine if historic breeding targets have resulted in changes in fructan levels and to identify potential breeding parents for the development of varieties with specific fructan levels. The impact of seasonal conditions on grain fructan levels were also investigated. Analysis of the varieties in this study indicated that historic breeding targets have not impacted on grain fructan levels. Fructan content in flours varied between 1.01 to 2.27%, showing some variation among the varieties. However, a significant variation in fructan levels was observed between different harvest years (mean values for 2015 and 2016 samples were 1.38 and 1.74%, respectively). While large variations in fructan contents of different varieties were not found, there were some varieties with consistently higher or lower fructan contents which could be used to breed varieties with specific fructan levels.