Anthracnose, caused by Colletotrichum species, can severely infect the fruits and leaves of more than 30 plants and thus results in great yield and quality losses. To identify the major Colletotrichum species infecting walnut fruits, strawberry leaves, grape fruits, and tea leaves in Shandong Province, China, 101 strains were collected and isolated. The morphological characteristics of all isolates were observed, and multilocus phylogenetic analyses (ITS, GAPDH, ACT, TUB2, CAL, CHS-1, and HIS3) were conducted on the representative isolates. The strains were identified as five Colletotrichum species, namely, C. gloeosporioides sensu stricto, C. fructicola, C. camelliae, C. acutatum sensu stricto, and C. viniferum. Among them, C. viniferum was reported for the first time from walnut fruits and strawberry leaves in Shandong Province, China. Corresponding leaves or fruits were used as a model to clarify the pathogenicity of these isolates. The results showed that C. fructicola obtained from strawberry leaves was more aggressive than C. viniferum. All of the isolates obtained from various hosts were highly sensitive to pyraclostrobin, difenoconazole, fludioxonil, tebuconazole, pyrisoxazole, and tetramycin in terms of mycelial growth inhibition (EC50 values of 0.07 to 1.63 mg/liter). The fastest mycelial growth was observed in the temperature range of 25-28°C for all isolates. In addition, anthracnose symptoms occur frequently under these conditions. Overall, this study can improve the understanding of Colletotrichum species causing anthracnose in walnut fruits, strawberry leaves, grape fruits, and tea leaves and can provide a solid foundation for the effective control of this disease in different hosts.