Phosphatidylinositol transfer protein (PITP) plays an important role in lipid metabolism and plant immune response, however its function is less well-known in sugarcane (Saccharum spp.). In this study, 51 SsPITP genes clustered into four groups (I, III, IV, and V), were identified in S. spontaneum. Results showed that expansion of the SsPITP gene family was mainly through whole genome/segmental replication and dispersed replication events, while purification selection was the main driving force for its evolution. Besides, the SsPITP gene family contained cis-regulatory elements related to light, hormone, and stress response in promoter regions. Interestingly, SsPITPs were differentially expressed under Sporisorium scitamineum stress. Furthermore, due to the continuous high expression of SsPITP29 in the sugarcane resistant variety after being infected with S. scitamineum, its homologous gene ScSEC14–1 with 99.69 % amino acid identity was cloned from the sugarcane hybrid cultivar. The results of transient expression in tobacco epidermal cells showed that the ScSEC14–1 protein was localized on the cell membrane and nucleus. Notably, overexpression of ScSEC14–1 increased the expression of phospholipase C genes in transgenic plants by activating the phosphatidylinositol/calcium/MAPK signaling pathways and plant hormone signal transduction, increasing chlorophyll content and catalase activity, reducing malondialdehyde content, thus enhancing disease resistance. This work will help to systematically understand the characteristics of sugarcane Sec14-like PITP gene family and the disease resistance function of the ScSEC14–1.