First Report of Colletotrichum fructicola Causing Anthracnose on Epimedium sagittatum (Sieb. et Zucc.) Maxim. in China

(Sieb.et Zucc.) Maxim., belonging to the family Berberidaceae and genus , is a perennial herb widely studied for its anti-osteoporosis, anti-cancer, and anti-sexual-dysfunction effects in Asian countries (Tan . 2016; Zhang . 2016). High levels of bioactive chemicals in spp. has endowed it with important clinical and commercial values (Liu . 2013). In September 2021, a leaf disease was found in Zhumadian City, China (32°58'12" N, 114°37'48" E). Survey statistics indicated that disease prevalence in a 266-ha planting area was approximately 29.6%. The lesions appeared at the leaf tips, gradually enlarged, and were brown with a yellow halo. Further, the lesions were dry with distributed black spots. Thirty infected leaves collected from five sites within the planting base . The collected leaves were cut into 5×5 mm pieces , surface-sterilized in 75% alcohol for 15 s, triple washed with sterile H O, disinfested with 0.1% HgCl solution for 30 s (Liu . 2021), triple washed again with sterilized H O, and then placed onto PDA and incubated in the dark for 3 d at 28°C. Subsequently, five fungal strains were purified; among them, only the isolate HY3-2 infected the host plant and was selected for further morphological characterization. The colonies of HY3-2 initially appeared white, their mycelia became gray at the center after 4 d, and orange-red conidial clumps appeared in them after 7 d. Conidia (10.0-19.5 μm × 4.5-5.6 μm, n=50) were single celled, nearly spherical or stick-shaped and colorless. Morphological characteristics of the isolate were consistent with those of species. Additionally, glycerol-3-phosphate dehydrogenase ( ), actin ( ), calmodulin ( ), β-tubulin 2 ( ), and chitin synthase-1 ( ), (Weir . 2012) were amplified and sequenced using the primers GDF/GDR, ACT-512F/783R, CL1C/CL2C, T1/Bt2b, and CHS-79F/354R, respectively for molecular identification. The resulting sequences were deposited in GenBank: (ON351609), (ON351608), (ON351610), (ON532788), and (ON532787). Phylogenetic analyses were performed by concatenating all the sequenced loci using the Bayesian method (Zhang . 2020). The phylogenetic tree showed that the isolate belongs to clade with a credibility value of 85%.To satisfy Koch's postulates, a conidial suspension (10 conidia/mL) of the isolate HY3-2 were prepared with sterile H O to infect the leaves. Ninety healthy leaves from 30 plants in pots were punctured using a sterilized needle (Huang . 2022), and inoculated by spraying the conidial