A protocol is described for high frequency plant regeneration from isolated leaf protoplasts of several genotypes of the wild tomato species Lycopersicon hirsutum f. hirsutum based on modified tomato protoplast culture methods. Three to four week old seedlings exposed to a low light pretreatment yielded protoplasts capable of sustained divisions on modified Lycopersicon Culture Medium. Plating efficiencies varied from 7.2%-25.9%. Colonies were transferred to modified solid greening medium after 25-35 days. Developing calli that turned bright green and produced dark green bud primordia were transferred to shoot induction medium. Shoot formation efficiencies ranged from 60%-85%. Shoots rooted easily and regenerated plants grown to complete maturity showed only transient somaclonal variation.