Candidatus phytoplasma affects a wide range of host plants. In recent years, the severity and distribution of the disease have increase many fold. The present study isolated the pathogen from leaves (chlorosis), dried branches, and the other tissues of the infected tree. Acacia trees (n=15) showing severe yellowing and desiccation in the landscape areas of Şanlıurfa province were sampled from the buds (V-shaped) in three different parts of the branches, backward from the top of the branches during the flowering period, and from the middle veins of the leaves on the same branch during the leafing period. Samples were initially amplified using the universal primer pair R16F1/R16R0 and subsequently subjected to nested PCR with the primer pair R16F2n/R2. The results indicated that, on average, 2 out of 15 buds collected from the apical parts of the branches, 11 out of 15 from the middle, and 6 out of 15 from the basal parts were infected with ‘Candidatus Phytoplasma australasia’. Moreover, all samples collected from the midribs of these 15 trees were found to be infected with Ca. P. australasia.