Bacterial and fungal contaminants of enset (Ensete ventricosum) cultures and microbes associated with surface-sterilized field material were identified by 16S/26S rDNA sequencing. Ten bacterial species were identified in 16 isolates from in vitro cultures and seven in 10 isolates from field clones. Three yeast species and one filamentous fungus were recorded as in vitro contaminants, whereas five yeast species were isolated from the field material. The bacterium, Pseudomonas reactans (6 isolates), and the yeast, Torulaspora delbrueckii (8 isolates), were the most frequent in vitro contaminants. Most of the bacterial species isolated from in vitro enset were Gram-positive and hitherto unrecorded as in vitro contaminants. The difficulty in controlling the in vitro contaminants is due to their apparent endogenous nature and their resistance to antimicrobial drugs.