Leaf stripe, caused by the fungus Pyrenophora graminea (Pg), is an important seed-borne disease of barley causing significant yield and quality losses worldwide. In this work, expression of phenylalanine ammonia-lyase (PAL) was monitored in infected leaves of two barley genotypes, Banteng (resistant) and Furat1 (susceptible) across four-time points of Pg systemic movement using qRT-PCR approach. Data showed that the systemic movement of the fungus was slower in the resistant cultivar than in the susceptible one, as confirmed by both microscopic examination and culturing on PDA media. qRT-PCR revealed that the expression patterns of PAL gene significantly increased in infected plants in comparison with non-infected controls. Its expression correlated with Pg systemic movement in the root, stem and leaf fractions of the resistant and susceptible barley cultivars. Taken together, based on the Pg systemic movement within barley plants with different resistance levels, our data strengthen the idea that PAL plays a role in barley leaf stripe reduction