In September 2015, post–harvest rot were found on a large number of kiwifruits in Hunan province of China which shows obvious brown disease spot. To figure the reason out, pathogen segregation and purification assay were conducted on the rotten parts of the kiwifruit. By morphological observation, pathogenicity verification, and molecular biological methods Phomopsis longicolla was finally identified as the cause of the kiwifruit rot.