Mitogen-activated protein kinase (MAPK) cascades are universal signal-transduction modules, but the available information is limited in pear (Pyrus). In this study, 87 MAPK genes were identified from five Rosaceae species: chinese white pear (Pyrus ×bretschneideri cv. Dangshansuli), peach (Prunus persica), apple (Malus domestica), strawberry (Fragaria vesca), and plum (Prunus mume), 23 of which came from chinese white pear, designated as PbrMAPK. Based on the phylogenetic analysis and the architectures of conserved protein motifs of these gene sequences, MAPK family genes of five Rosaceae species were classified into two primary types (I and II) or four groups (Classes A–D). We have indicated that both segment and tandem duplications significantly contributed to the expansion of the MAPK family in Rosaceae by analysis of genomic evolution. In chinese white pear pollen, the expression analysis revealed that all PbrMAPKs could respond to temperature stresses (high/low temperature) and phytohormones, except PbrMAPK8 and PbrMAPK19 that displayed lower expressions, which suggested that PbrMAPKs play pivotal roles in signal-transduction pathways. In addition, we determined that PbrMAPK13 is located in the nucleus and plasma membranes. The lengths of pollen tubes became shorter when PbrMAPK13 was silenced by antisense oligonucleotide transfection. Our results provided an evolutionary foundation and functional characterization for MAPK gene families in chinese white pear and other plant species so as to elucidate their biological roles.