Whitefly, Bemisia tabaci, is one of the most important pests of tomato, Solanum lycopersicum L. The Mi-1 gene mediates tomato resistance to the Middle East-Asia Minor 1 (MEAM1) and Mediterranean (MED) species of B. tabaci, three species of root-knot nematodes, Meloidogyne spp., and the potato aphid, Macrosiphum euphorbiae. Tomato seedlings bearing the Mi-1 gene are resistant to nematodes soon after germination but resistance to aphids is developmentally regulated; a reliable conclusion about Mi-1 resistance to B. tabaci was not available to date. In the present work, 3-, 5- and 8-week-old plants of the tomato cultivars Motelle and Moneymaker (bearing and lacking the Mi-1 gene, respectively) were simultaneously tested under free-choice (antixenosis) and no-choice (antibiosis) conditions, to assess the real influence of plant age on the Mi-1-mediated resistance to the MED species of B. tabaci. Subsequently, plants of the same age but with different level of development were compared to check whether the plant size can also affect this tomato resistance. Obtained results demonstrated that Mi-1-mediated resistance to B. tabaci is developmentally regulated, as variations in the age of bearing-Mi-1 plants affects most infestation parameters tested. Differences between cultivars with and without the Mi-1 gene were significant for 8- but not for 3-week-old plants. For 5-week-old plants, differences between cultivars were less pronounced than in older plants, expressing an intermediate level of resistance in Motelle. Plant size also influenced whitefly infestation and reproductive activity on the resistant cultivar. However, plant age has more impact than plant size on the Mi-1-mediated resistance of tomato to B. tabaci.