GPCR is the abbreviation of G protein coupled receptor. As a kind of important cell surface receptor with seven transmembrane helical structure, GPCR plays an important role in cellular signal transduction in biology. However, the physicochemical properties and genetic relationship of candidate GPCR proteins of Colletotrichum graminicola are not clear, which restricts the study of the pathogenic mechanism of this strain. In this study, based on the 220 candidate GPCR protein sequences of C. graminicola, the signal peptide, physicochemical properties and hydrophobicity of the above proteins were identified by online prediction programs such as SignalP5.1, ProtParam and ProtScal. The results showed that there were only 7 candidate GPCR proteins containing signal peptide, and the theoretical isoelectric points were mainly more than 6.01, most of which were neutral or basic proteins, all of which were hydrophobic proteins. At the same time, the genetic relationship of the above proteins was analyzed by MEGA software, and it was clear that the candidate GPCR proteins were mainly divided into A, B and C categories, which were closely related to their conserved domains. This study provides an important theoretical support for further functional analysis of GPCR protein of C. graminicola, and also provides an important theoretical basis for the study of the function of GPCR protein in other Colletotrichum species.