Abstract
MicroRNAs (miRNAs) are key regulators of gene expression in plants, influencing various biological processes such as oil quality and seed development. Although, our knowledge about miRNAs in olive (Olea europaea L.) is progressing, with several miRNAs being identified in previous studies, but most of these reported miRNAs have been predicted without the aid of a reference genome, primarily due to limited genome accessibility at the time. However, significant knowledge gaps still need to be improved in this area. This study addresses the complexities of miRNA detection in olive, using a high quality reference genome and a combination of genomics and machine learning-based methods. By leveraging random forest and support vector machine algorithms, we successfully identified 56 novel miRNAs in olive, surpassing the limitations of conventional homology-based methods. Our subsequent analysis revealed that some of these miRNAs are implicated in the regulation of key genes involved in oil quality. Within the context of oil biosynthesis pathways, the novel miRNA Oeu124369 regulates fatty acid biosynthesis by targeting acetyl-CoA acyltransferase 1 and palmitoyl-protein thioesterase, thereby influencing the production of acetyl-CoA and palmitic acid, respectively. These findings underscore the power of machine learning in unraveling the complex miRNA regulatory network in olive and provide a high quality miRNA resource for future research aimed at improving olive oil production by exploring the target genes of the identified miRNAs to understand their role and their biological processes.