Abstract: Serine/arginine-rich (SR) protein, as a splicing factor, plays a crucial role in many physiological and biochemical processes, especially in precursor mRNA splicing and mRNA metabolism. Based on two typical SR protein in Schizosaccharomyces pombe, to search SR protein sequence from the protein database of Colletotrichum graminicola with Blastp and keywords, two SR proteins CgSrp1 and CgSrp2 were identified in Colletotrichum graminicola. CgSrp1 and CgSrp2 contain the N-terminal RNA Recognition Motif domain and serine/arginine dipeptides towards the C-terminus with SMART analysis. BLASTp in NCBI was used to perform homology analysis and construct phylogenetic trees with other species. Sequence variation of SR proteins was discovered between plants and fungi by amino acids alignment. Meanwhile, bioinformatics analysis on SR protein was made including the physical and chemical properties, hydrophobicity, subcellularlocalization, the secondary and tertiary structure. It is showed that CgSrp1 and CgSrp2 were hydrophilic proteins, both of them located in the nucleus and contained α-helixs, β-sheets and random coil. CgSRP1 and CgSRP2 gene fragments were successfully cloned using wild-type Colletotrichum graminicola CgM2 DNA as template, indicating the presence of CgSRP1 and CgSRP2 genes in CgM2.