Diseases affecting strawberries have been of major concern in recent years because of their widespread occurrence and potential for yield loss. Anthracnose caused by the fungus Colletotrichum acutatum is one of the most serious disases of strawberry worldwide. Although chemical controls are being used to treat anthracnose, generating disease resistant plants is a more attractive solution to the problem because chemicals can pose a health hazard, have a negative impact on the environment and may only be moderately effective. Tissue culture-induced (somaclonal) variation provides us with one strategy for generating disease-resistant genotypes. An in vitro screening system was used to evaluate several commercially important cultivars, Chandler, Delmarvel, Honeoye, Latestar, Pelican and Sweet Charlie, and shoots regenerated from leaf explants of these cultivars for resistance to C. acutatum isolate Goff (highly virulent). Somaclones with increased levels of anthracnose resistance were identified for all the cultivars. The greatest increases in disease resistance were observed for somaclones of cultivars Chandler, Pelican and Sweet Charlie that exhibited 6.8-, 12-, and 4.2-fold increases in resistance, respectively. These studies provide evidence that: 1) in vitro screening can be used to evaluate strawberry germplasm for anthracnose resistance, 2) soma-clonal variation is influenced by stawberry genotype, and 3) generating somaclonal variants may be a feasible approach to obtaining strawberry plants with increased levels of anthracnose resistance.