Anthracnose is a devastating fungal disease in strawberry nurseries. Multiple Colletotrichum species are responsible for strawberry anthracnose. In this study, 105 Colletotrichum isolates were obtained from strawberry seedlings with anthracnose symptoms in fifteen nurseries located in Zhejiang province, China, and were classified based on multilocus sequence and morphological characteristic analyses. Analysis of ITS, ACT, CAL, CHS-1, and GAPDH revealed that four species within C. gloeosporioides species complex, including C. siamense (56 isolates, 53.3%), C. fructicola (37 isolates, 35.2%), C. gloeosporioides (7 isolates, 6.7%), and C. aenigma (5 isolates, 4.8%), were detected in diseased seedlings. Thiophanate-methyl is one of the benzimidazole fungicides, and has long been used to control strawberry anthracnose in China. Here, thiophanate-methyl resistance of Colletotrichum isolates was determined by the minimum inhibitory concentration (MIC) method. Our results indicated that the resistance frequency was up to 96.2%, containing 94.3% of highly resistant isolates. Only four sensitive isolates (two C. fructicola, one C. gloeosporioides, and one C. siamense isolates) and two moderately resistant isolates (one C. aenigma isolate and one C. siamense isolate) were detected. Our data indicated that the high resistance was mainly caused by the E198A mutation in the β-tubulin protein. In addition, F200Y (TTC→TAC) in the β-tubulin protein were detected in two moderately resistant isolates. Based on the point mutation at codon 198 (GAG→GCG) in the β-tubulin gene of Colletotrichum isolates, we developed a loop-mediated isothermal amplification (LAMP) assay to rapidly detect the E198A mutants. Collectively, our study indicated that four species within the C. gloeosporioides species complex were associated with anthracnose symptoms in strawberry nurseries in Zhejiang province, and serious resistance was widespread in each Colletotrichum species.

Anthracnose is a fungal disease that seriously threatens grape production and quality. Multiple Colletotrichum species are detected in anthracnose grapes in vineyards. In this study, diseased grapes were collected in four counties in Zhejiang, and 43 Colletotrichum isolates were obtained. Multi-genes (ITS, TUB2, ACT, CHS-I, and GAPDH) and morphological characteristic analyses showed that C. fructicola (40 isolates, 93%), C. aenigma (two isolates, 4.7%), and C. pseudoacutatum (one isolate, 2.3%) were the cause of grape anthracnose in Zhejiang. Among the three Colletotrichum species, C. fructicola was the prevalent and dominant species in all sampled counties; C. pseudoacutatum was first identified as the pathogen responsible for grape anthracnose. There were significant differences in the sporulation among the three Colletotrichum species, as well as in the spore germination. Pathogenicity testing showed that all species can infect grapes, resulting in anthracnose. On the other hand, the virulence of species was varied and may be associated with their spore germination. This is the first study to characterize the Colletotrichum species causing grape anthracnose in Zhejiang Province and reveal that C. fructicola is the dominant species. The determination of Colletotrichum species associated with grape anthracnose may contribute to the study of epidemiology and development of an efficient strategy for controlling anthracnose in the vineyards.