Gene expression studies by real-time PCR constitute a powerful tool to analyze the mechanisms underlying plant abiotic-stress tolerance. One of the crucial steps of this technique is the selection and validation of reference genes to normalize target gene expression under different stress conditions. In an attempt to find the best reference genes for wheat root salinity research, we evaluated the stability of the three most commonly used reference genes (GAPDH, Actin and Ta.22845) using the NormFinder, BestKeeper statistical algorithms and the comparative ΔCT method. The results indicated that Actin was the most suitable reference gene for gene expression normalization under salinity treatment in wheat root.