First Report of Trichothecium roseum Causing Postharvest Fruit Rot on Purple Passion Fruit in China.

Purple Passion fruit (Passiflora edulis) is widely cultivated in many regions of southern China as an edible tropical fruit with excellent nutritional value and high economic value. In July 2021, postharvest fruit rot was observed on 20-25% of purple passion fruit in several fruit markets of Dehong City in Yunnan Province. Symptoms on infected fruits were irregular, pink-brown, soft, and water-soaked lesions, which enlarged and formed sunken patches with time as well as producing a small amount of white mycelium. To isolate the causal organism, five diseased fruits were collected from different fruit markets. A conidial mass from an individual sorus observed on an infected fruit was isolated and cultured on potato dextrose agar (PDA) supplemented with 50 μg ml-1 of streptomycin, and five fungal isolates were obtained. These isolates were morphologically similar and produced pale pink colonies on PDA for 7 days containing several conidiophores with abundant conidia. Mycelia were hyaline, 2 µm in diameter, and conidiophores were simple or branched (100 to 286 × 1.5 to 2.5 µm, n=50). Conidia were pyriform, ovate, with papillary protuberances at one end. Almost all conidia were two-celled and single-septate (5.8 to 9.1 × 1.7 to 4.9 µm, n=50). The morphology of the fungi resembled Trichothecium roseum as reported previously (Inácio et al. 2011). To further confirm the fungal species, isolate PASF4 was selected for molecular identification by amplifying and sequencing the ribosomal internal transcribed spacer (ITS) and large subunit (LSU) genes. Primers and PCR amplification were described by Fell et al. (2000). Results showed that both the ITS (GenBank accession OL336243) and LSU (OL336242) gene sequences had 100% similarity to T. roseum in NCBI database (MH856757 and MH868278). Maximum likelihood tree was constructed using MEGA 7 (Felsenstein, 1981) based on concatenated sequences (ITS and LSU) of isolate PASF4 and reference strains. Phylogenetic analysis showed that isolate PASF4 belonged to T. roseum clade. Based on morphological characteristics and phylogenetic analysis, isolate PASF4 was identified as T. roseum (Inácio et al. 2011). To confirm their pathogenicity, healthy purple passion fruits (cv. Tainong-1) were disinfected in 0.5% NaClO solution for 2 min, and then washed with sterile water. After wounding with a sterile needle, the fruits were inoculated by placing mycelium agar plugs on the wounds, and mock inoculation with mycelium-free PDA plugs served as control. Five fruits were used in each treatment. All fruits were maintained in plastic boxes at 25 °C. Disease symptoms appeared after inoculation for 4-7 days on all inoculated fruits, which were similar to those observed in fruit markets. No symptoms were observed on fruits used as control. The Trichothecium isolates were re-isolated from symptomatic fruits thus fulfilling Koch's postulates. Trichothecium roseum has been reported to cause fruit rot of tomato, apple and orange in Pakistan (Hamid et al., 2014) and fruit rot of pepper in China (Lin et al., 2016). To our knowledge, this is the first report of T. roseum causing fruit rot on purple passion fruit worldwide, and these data will provide useful information for developing effective control strategies.