The objective of this study was to develop resistance in vitro against Fusarium equisetii in Robinia pseudoacacia L. Callus was induced from cotyledon explants of R. pseudoacacia, a leguminous landscaping tree, on solid Murashige and Skoog (MS) medium supplemented with different concentrations and combinations of 1-naphthalene acetic acid (NAA; 0.1-0.5 mg/l) and N 6 -benzyladenine (BA; 0.1-0.3 mg/l). Among them, 1.0 mg/l NAA and 1.0 mg/l BA was the best combination for callus induction, which was friable and light brown. Callus fresh weight increased and turned pale yellow after subculture. The effect of different concentrations of a toxic culture filtrate of Fusarium equisetii (Corda) Sacc. was investigated by plating friable callus on selective medium. Resistant calli were selected at 10% of the culture filtrate on which 15.33% calli survived, although no callus formed at higher concentrations of toxic culture filtrate. Shoots, which regenerated from resistant calli on MS medium supplemented with 0.05 mg/l NAA and 0.6 mg/l BA, could root on half-strength MS medium containing 0.02% activated charcoal. Selected resistant plantlets, which survived on 2.5-10% of the culture filtrate compared to the control, showed reduced wilt incidence (approximately 70%) indicating a resistant response in tissues towards F. equisetii .