Phospholipase is a class of phospholipase hydrolases and signal transducers that play an important role in regulating plant growth and development and stress response. However, the function of phospholipase A1 (PLA1) in plant response to biological and abiotic stresses is rarely reported. In this study, a subtype of phospholipase PLA1, MaPLA1-2D was cloned in mulberry, and the sequence feature, tissue expression, stress-induced expression of MaPLA1-2D, and subcellular localization of the protein were analyzed. The results showed that MaPLA1-2D contains 4 isoforms, named MaPLA1-2D.1- MaPLA1-2D.4. The four genes were highly expressed in the roots and leaves of mulberry, and the proteins were mainly localized in chloroplasts. The MaPLA1-2D members had high sequence similarity and close evolutionary relationship with Arabidopsis AtDAD1 in the conserved domain. It was showed that the promoters of the four MaPLA1-2D genes contained a variety of stress-responsive cis-elements and hormone responsive elements, and the gene expressions of MaPLA1-2D were significantly induced by drought stress and abscisic acid (ABA) treatments. These results indicate that MaPLA1-2D genes are homologous to Arabidopsis DAD, which may play an important role in abiotic stress response of mulberry.