Characterization of the miR165 family and its target gene Pp-ATHB8 in Prunus persica

A study to identify and validate Prunus persica microRNAs belonging to the ppe-miR165 family was conducted using miR-RACE where the expression of these miRNAs could be detected in leaves, flower buds, flowers and fruits of P. persica by quantitative real-time PCR (qRT-PCR) with some members of this miRNA family exhibiting tissue-specific expression. In addition, Pp-ATHB8 which is one of the potential target genes for ppe-miR165 family was experimentally verified by PPM-RACE and RLM-RACE, from which the cleavage sites of the target mRNA were mapped and the expression patterns of cleaved fragments also detected, thus demonstrating the mode by which ppe-miR165 regulates the target gene Pp-ATHB8. Furthermore, spatiotemporal expression levels of the target gene and its cleaved fragments were analyzed by qRT-PCR, where they were found to exhibit expression trends at variance from ppe-miR165, thus indicating the cleavage mode of this miRNA on its target gene. The characterization of this important miRNA and its interaction with an equally important target gene in P. persica, further deepens our understanding of the role of ppe-miR165 in P. persica and expands the knowledge of small RNA-mediated regulation in this fruit crop.

► We predicted and isolated members of miR165 family from Prunus persica.
► We predicted and characterized Pp-ATHB8, a ppe-miR165 target gene in peach.
► We validated ppe-miR165 family members and its target gene Pp-ATHB8.
► Expression of ppe-miR165 members and Pp-ATHB8 in tissues of peach revealed.
► The mode by which ppe-miR165 regulates Pp-ATHB8 was demonstrated.