Fusarium head blight (FHB) is one of the most studied fungal diseases of wheat, causing massive grain yield and quality losses. This study aimed to extend previous studies on the physiological and biochemical responses of winter wheat to FHB stress in a controlled environment by focusing on the ascorbate-glutathione pathway (AsA-GSH), photosynthetic efficiency, and stress hormone levels, thus providing insight into the possible interactions of different defense mechanisms during infection. The activity of AsA-GSH metabolism was increased in FHB resistant varieties, maintaining the redox state of spikes, and consequently preserving functional photosystem II. Furthermore, carotenoids (Car) were shown to be the major pigments in the photosystem assembly, as they decreased in FHB-stressed spikes of resistant and moderately resistant varieties, compared to controls. Car are also the substrate for the synthesis of abscisic acid (ABA), which acts as a fungal effector and its elevated content leads to increased FHB susceptibility in inoculated spikes. The results of this study contributed to the knowledge of FHB resistance mechanisms and can be used to improve the breeding of FHB resistant varieties, which is considered to be the most effective control measure.