In order to screen out the essential oils with better inhibitory effect on grape Fusarium sp., vitro assay was used to measure the growth of grape Fusarium sp. colonies. The results showed that when the concentration of essential oils was 1000 μL/L, among all 21 essential oils, six essential oils could completely inhibit the growth of Fusarium sp. When the concentration of these six essential oils was reduced to 500 μL/L, only basil essential oil had sustained antifungal effect, but without continuous inhibitory effect at 250 μL/L. Thus, basil essential oil has sustained antifungal effect on grape Fusarium sp. in vitro assay, but its concentration should be higher than 250 μL/L.