Aggressiveness variation among 16 isolates of four Fusarium species (F. culmorum, F. solani, F. verticillioides and F. equiseti) causing Fusarium head blight (FHB) was studied in vitro. Evaluation of three aggressiveness criteria involved in a Petri-dish test: germination rate reduction, standardized area under disease progress curve (AUDPCstandard), and coleoptile length reduction was carried out on the barley cultivar Arabi Aswad. Results showed differences between barley plants inoculated with FHB isolates and control for the three tested aggressiveness criteria. Regarding AUDPCstandard and Petri-dish aggressiveness index which is calculated from the mean value of three aggressiveness criteria, within and among variation was detected. Intra- and inter-species variability was not distinguished for the other two aggressiveness criteria. However, pathogenic level observed among 16 isolates can not be differentiated within the four FHB species. Significant correlation was detected only between AUDPCstandard and Petri-dish aggressiveness index. The results were comparable with those previously obtained using the same fungal isolates on wheat cultivar in vitro. It seems that FHB isolates recovered from wheat spikes and tested on wheat plants showed a similar range of aggressiveness on a barley cultivar, Arabi Aswad.