During the winter of 2018, leaf blight on florist’s daisy (Chrysanthemum morifolium L.) was noticed in Egypt. The disease, which was identified as caused by Alternaria alternata, was widely spread and led to serious damage for the exportation sector of this crop. Therefore, a study was conducted to better understand what can be conducted to minimize the problem in the future. Isolates were gathered and evaluated on five chrysanthemum cultivars (i.e., ‘Feeling Green Dark’, ‘Talitha’, ‘Chrystal Regan’, ‘Arctic queen’, and ‘Podolsk Purple’) grown in a greenhouse. The objectives were to isolate and identify the phytopathogen and detect the resistant degree of these cultivars with emphasis on the early growth stages of the crop. The results showed that ‘Podolsk Purple’ was the most resistant cultivar against the different isolates during the rooting and seedling growth stages. ‘Chrystal Regan’ was very susceptible to the different isolates. In addition, the isolate from ‘Feeling Green Dark’ was the strongest, which negatively affected the chlorophyll content and its fluorescence parameters besides other measured vegetative and anatomical features. The findings indicated that the best anatomical characters of the stem and leaf, like the thickness of cuticle and cortex, stem diameter, xylem vessel diameter, and thickness of epidermis as well as lamina thickness were recorded in the ‘Podolsk Purple’ cultivar. This study highlighted that by using the right cultivars, chrysanthemum can be cultivated during the winter season under Egyptian conditions. These results can be a part of solution to overcome the leaf blight caused by A. alternata on chrysanthemum during the early growing stages.