Abstract: Foliar phase of anthracnose, caused by Colletotrichum sublineolum is the most important leaf disease of sorghum. Due to the hyper-variable nature of the fungus, continuous evaluation of sorghum germplasm to identify new sources of resistance is imperative. Field and greenhouse evaluations for anthracnose resistance, especially with large numbers of sorghum lines/accessions can be expensive, time consuming and require large spaces and labor. In this study, 16 sorghum lines were evaluated by putting a drop of the mixture of C. sublineolum isolates suspension on each side of the midrib of adaxial excised leaves plated on half-strength potato dextrose agar medium and concurrently as whole plants inoculated with a mixture of C. sublineolum isolates-colonized grain and conidial suspension in the greenhouse. Each line exhibited the same reaction when challenged with C. sublineolum either using the excised leaf assay or screened in the greenhouse, indicating that the excised leaf assay is as effective in identifying susceptibility or resistance to the anthracnose pathogen. In both screening methods, SC748 was the only resistant line. The excised leaf assay was completed in 4 days while the greenhouse evaluation was arrested 44 days post-inoculation. Thus, the excised leaf method could offer several advantages in screening sorghum for anthracnose resistance such as reducing the time for conducting the experiment, labor, space and increasing the number of isolates that can be tested within a short period.