Members of Yellow Stripe-Like (YSL) family of proteins are membrane bound oligopeptide transporters involved in transport of metals, and remobilization of nutrients in storage tissue. The evolution of this gene family in plants is not well understood. Here we have analysed 199 YSLs from 25 different plant species for intron-exon structure, gene duplication, phylogenetic relationship, conserved motif and interaction network towards exploring the gene duplication pattern, conservation and evolution. The YSLs belonging to 25 species formed two major groups and further divided into ten sub-groups based on phylogenetic analysis. Conserved motif analysis of all the YSLs revealed the highly conserved nature of the transmembrane protein among the 25 species. Further, terminal and non-terminal duplication was found to play a major role in the expansion of the YSL family. We also analysed the transcriptome data under iron (Fe) deficiency condition in rice root. Three rice YSL genes, OsYSL15, OsYSL13 and OsYSL9 were found to be up-regulated under Fe deficiency stress. Functional network analysis of rice YSLs revealed 48 interactions involving 21 genes which were mostly Fe, zinc (Zn) stress related proteins such as OsNRAMP, OsIRO2, OsNAS1, OsNAS2, OsIRT1, OsIRT2 and OsDMAS1. Highly conserved nature of YSLs provides an opportunity for extrapolation of functionality of YSL proteins from rice to other monocot species of the Poaceae family.