The whitefly leads extensive damage to plants through direct feeding, honeydew secretion, plant physiological disorders, and vectoring plant viruses. This study aimed to evaluate the physiological characteristics of susceptible and resistant soybean cultivars to B. tabaci. The experiments were conducted in a greenhouse. Eleven soybean cultivars were selected and infested with 100 adults of B. tabaci at the V3 stage. The evaluation of photosynthetic parameters, such as photosynthetic rate, leaf transpiration, stomatal conductance, and internal CO2 concentration, revealed that B. tabaci infestation influenced gas exchange in soybean plants. The photosynthetic rate was higher in cultivars AS3810 and M8349 during the V6 stage. Infestations led to alterations in photosynthetic parameters, suggesting increased energy demand to maintain photosynthetic activity. However, the response to infestation varied among different cultivars, indicating varying levels of resistance and tolerance to the whitefly's damage. Additionally, the impact of infestation was more significant during the vegetative phenological. In conclusion, B. tabaci infestation affects soybean plants' physiology, leading to changes in gas exchange parameters and water use efficiency. The response to infestation varied among soybean cultivars, suggesting potential differences in resistance to the pest. The study highlights the importance of evaluating the physiological impact of whitefly infestations on soybean.