Two synthetic brassinosteroids, 24-epibrassinolide (24-epiBR) and 2o,3a,l7P-trihydroxy-5cu-androstan-6-one (THA-BR), exhibit different effects on growth of tobacco callus tissue. When added to a culture medium containing growth-limiting amounts of auxin, 24-epiBR reduced and THA-BR increased the fresh weight yield of tissue up to 53% and 207%, respectively, after 6 weeks of cultivation. The stimulatory and inhibitory effects of the two brassinosteroids on tissue growth occurred over a broad range of concentrations without a pronounced maximum corresponding to the 'yes or no' type of response. Different effects of 24-epiBR and THA-BR on tissue growth were inversely proportional to the content of endogenous cytokinins. Maximum contents of predominant cytokinins N6-(A=-isopentenyl)adenine (iP) and trans-zeatin (Z) in tissues supplied with 24-epiBR in growth-inhibiting concentrations were up to 3.7 fold and 3.4 fold higher, respectively, as compared to tissues grown on media containing growth-stimulating concentrations of THA-BR. Stimulation of tissue growth by THA-BR correlated with content of endogenous IAA and an inverse correlation was found between the content of endogenous IAA and cytokinins in tissues supplied with 24-epiBR. THA-BR exhibited weak cytokinin-like activity in a bioassay based on stimulation of growth of lateral buds of pea while 24-epiBR was inactive. Results indicate that the qualitatively different effects of the two brassinosteroids on growth of tobacco tissue may reflect their different influence on content of endogenous cytokinin.Abbreviations: BR(s) = brassinosteroid(s); 24-epiBR = 24-epibrassinolide; THA-BR = 2a,3a,l7/3-trihydroxy-5cu-androstan-6-one; CK(s) = cytokinin(s); iP = N6-(A=-isopentenyl)adenine; [9R]iP = N6-(A=-isopentenyl)adenosine; Z = trans-zeatin, [9R]Z = ribosyl-trans-zeatin; ABA = abscisic acid; IAA = indole-3-acetic acid; NAA = naphtalene-1 -acetic acid; DEAE cellulose = diethylaminoethyl cellulose; HPLC = high performance liquid chromatography; ELISA = enzyme linked immuno-sorbent assay