Effect of auxins: (NAA, IAA), cytokinin BAP and culture conditions (light, darkness) on callus induction in cotyledons of tomato cultivar ‘Maskotka’ and the wild form of Lycopersicon peruvianum was investigated. Callus was obtained in all experimental combinations, except of the culture with L. peruvianum on the medium with 1.0 mg⋅dm⁻³ of IAA. Callus weight, colour and structure depended on the tomato genotypes and experimental conditions. Best medium for the culture of tomato cultivar ‘Maskotka’ and the wild form L. peruvianum proved to be MS medium supplemented with 2.0 mg⋅dm⁻³ of IAA and 1.0mg⋅dm⁻³ of BAP.