In the human body, about 53% of Mg is involved in the development and maintenance of bone and other calcified tissues, although it also has a physiological role in protein synthesis, muscle and nerve functions, blood glucose control and blood pressure regulation. Nevertheless, Mg deficiency triggers electrolyte disturbance that can result in multiple symptoms, namely, tremors, poor coordination, muscle spasms, loss of appetite, personality changes, and nystagmus. Complications may include seizures or cardiac arrest. To surpass Mg deficiency, biofortification is a strategy that can boost nutrient enhancement in food crops and can increase nutrient uptake and accumulation in the human body. Accordingly, this study aimed to develop a technical itinerary for Mg biofortification in Lycopersicum esculentum variety H1534. Tomato biofortification was promoted during the respective life cycles throughout six leaf applications with two different treatments (4% and 8%) of MgSO4, equivalent to 702 and 1404 g ha−1. At harvest, the biofortification indexes of Mg were 2.01- and 1.71-fold higher (after spraying with 4% and 8% MgSO4, respectively), with synergistic trends found only with Zn and Fe, whereas P did not vary significantly among treatments. Among treatments, relevant deviations could not be found for total soluble solids, height, diameter and color; however, minor changes in dry weight were detected. It can be concluded that the Mg biofortification of tomato variety H1534 can be performed to add nutritional value to tomato-based processed food products.