Tomato (Solanum Lycopersicum L.), the most important vegetable in Colombia and worldwide, is attacked by numerous diseases, among them, the vascular wilting caused by Fusarium oxysporum f. sp. lycopersici. The disease can cause losses between 21 and 47% in full exposure and under cover crops. The objective of this work was to compare four inoculation methods of this fungus in seedlings of the Sweet Million cultivar obtained from seeds and planted in sterilized Canadian peat and in MS medium and maintained under controlled conditions at the Laboratory of Tissue Culture and thermotherapy room of the Universidad de Caldas where 25 seedlings were selected per method of inoculation. The treatments were: Injection in the crown root (T1), immersion of the roots (T2), agar disc in the crown root (T3), and inoculation in vitro (T4). The final disease incidence and the comparative tests of Duncan showed highly significant differences (p≤0.001) among the different methods of inoculation. The only method that induced a 100% incidence of the vascular wilt in the shortest period of incubation was the in vitro method. The highest rate of disease development (r) was obtained with the in vitro inoculation method (r=0.92) followed by agar discs, immersion, and injection. The in vitro inoculation method can be useful for the massive selection of tomato materials promising against vascular wilt. © 2019. Acad. Colomb. Cienc. Ex. Fis. Nat.