Waterless transport is an alternative method for live aquatic products. However, the changes that occur in the fish body after transportation are not known. Herein, the effects of different transport methods on quality of turbot (Scophthalmus maximus) were investigated. Fish were divided into four groups: Group A (water, 17°C), Group B (waterless, 3°C), Group C (water, 3°C), and Group D (control, 17°C). In the case of 100% survival, the maximum transport time was 40 h for Group A, 55 h for Group B, and 90 h for Group C. Both crude protein and crude fat decreased significantly (p < 0.05) in the treatment groups compared with the control group. Compared with control group, lactic dehydrogenase (LDH), lactic acid (LA), and serum cortisol (COR) increased significantly (p < 0.05) in all treatment groups. After fish recovered, LDH, LA, and COR values were all significantly (p < 0.05) lower than their corresponding non‐recovered group. Compared with the control group, the expression of proteins related to muscle motility and stress capacity both decreased in the low‐temperature, waterless group. Together, obvious damage may occur to flesh quality, liver, and heart of turbot when the transport time is >40 h, regardless of the transport method.