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Application of date palm waste compost is quite beneficial in improving soil properties and crop growth. However, the effect of its application on soil microbial communities is less understood. High-throughput sequencing and quantitative real-time PCR (qPCR) were used to evaluate the effect of compost application on the soil microbial composition in a barley field during the tillering, booting and ripening stages. The results showed that compost treatment had the highest bacterial and fungal abundance, and its application significantly altered the richness (Chao1 index) and α-diversity (Shannon index) of fungal and bacterial communities. The dominant bacterial phyla found in the samples were <i>Proteobacteria</i> and <i>Actinobacteria</i> while the dominant fungal orders were <i>Ascomycota</i> and <i>Mortierellomycota</i>. Interestingly, compost enriched the relative abundance of beneficial microorganisms such as <i>Chaetomium</i>, <i>Actinobacteriota</i>, <i>Talaromyces</i> and <i>Mortierella</i> and reduced those of harmful microorganisms such as <i>Alternaria</i>, <i>Aspergillus</i> and <i>Neocosmospora</i>. Functional prediction based on Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt) showed that amplicon sequence variant (ASV) sequences related to energy metabolism, amino acid metabolism and carbohydrate metabolism were associated with compost-treated soil. Based on Fungi Functional Guild (FUNGuild), identified fungi community metabolic functions such as wood saprotroph, pathotroph, symbiotroph and endophyte were associated with compost-treated soil. Overall, compost addition could be considered as a sustainable practice for establishing a healthy soil microbiome and subsequently improving the soil quality and barley crop production.