γ-Aminobutyric acid (GABA), which displays anti-hypertensive and diuretic effects, is known to be produced in germinated wheat. The effect of sequential hydration combined with anaerobic and heat treatment on the GABA content in wheat was investigated. The GABA content in soft white winter (SWW) and dark northern spring (DNS) wheat was 0.80 and 1.18mg/100g, respectively. Both the anaerobic and the heat treatment contributed to the increase of GABA in SWW and DNS wheat. The GABA contents in SWW and DNS wheat was increased by germination for 48h after sequential hydration with anaerobic and heat treatment at a target moisture content of 35% to 45.65 and 47.40mg/100g, respectively. Our results suggest that combined anaerobic and heat treatment after sequential hydration may be used for the preparation of wheat with high GABA content, which can then be used as a natural resource of functional foods.