Calli have been initiated in vitro from young internodes (control and rubbed) of Bryonia dioica, where previously it had been shown, using intact plants, that rubbing induced limited growth through enhanced lignification. Calli derived from rubbed internodes were somewhat more compact and showed biochemical changes, i.e. enhanced activity of total peroxidase and isoperoxidases, enhanced production of l-aminocyclopropane-l-carboxylic acid (ACC) and ethylene, enhanced tissue capacity to convert ACC into ethylene, enhanced activity of phenylalanine ammonia-lyase (PAL) and higher content of lignin, which characterized rubbed internodes. Differences in ethylene metabolism between the two types of calli tended to fade from the third week onwards of initial culture, whereas lignin content, peroxidase activity and peroxidase isoenzyme pattern appeared to be more persistant rubbinginduced markers for several subcultures. The results point to the persistance of environmentally induced changes in gene expression.