First report  flat peach fruit rot caused by Fusarium fujikuroi in China.

Flat peach (Prunus persica L. Batsch. var. compressa Bean) is a distinctive peach variety with flat shape. It is well known for its high nutritional value and pleasant flavor, and has become the primary cultivar in Shengzhou city, Zhejiang province, China. In July 2021, we discovered that 8%-10% of flat peach (data gained from a survey of fifteen trees in each orchard) fruits in three orchards (about 2.6 ha) had a fungal disease in Jinting Town, Shengzhou. The infected fruit by this pathogen showed soft, brown, and sunken lesions, accompanied by a sour odor and white mycelia. For pathogen isolation, ten infected fruits were surface sterilized (75% ethanol for 30 s and 2.0% NaClO for 3 min), then rinsed with sterile distilled water three times. The tissues from the margin of lesions were cut into 0.6×0.6 cm pieces and transferred to potato dextrose agar (PDA) medium containing 30.0 μg/mL chloramphenicol. The plates were incubated at 25 °C for 5 days in the dark. Mycelia growing from tissues were subcultured onto fresh PDA medium to get a pure isolate, which formed dense white hyphae after 4 days. The average growth rate of mycelium on PDA medium was 3.4 ± 0.2 mm/d, and the colonies were pale purple after 5 days. Macroconidia was slender, slightly curved, almost 2 to 3 septa, with a bend and tapering apical cell and poorly developed foot cell, and the size was 4.5 to 16.9 × 1.8 to 4.0 μm (n= 50) μm. No microconidia and chlamydospores were observed. This isolate's morphological and cultural characteristics were close to Fusarium fujikuroi (Leslie and Summerell. 2006). To further get the phylogenetic evidence, the nuclear ribosomal internal transcribed spacer region (ITS) and translation elongation factor-1 (TEF-1α) genes of three representative isolations were amplified by primers ITS1 (F:5'-GGAAGTAAAAGTCGTAACAAGG-3') /ITS4 (R5'-TCCTCCGCTTATTGATATGC-3') (Groenewald et al. 2013), and EF1-728F (5'-TACAARTGYGGTGGTATYGACA-3')/ EF1-986R (5'-ACNGACTTGACYTCAGTRGT-3') (Carbone and Kohn. 1999), respectively. The amplified sequences were submitted to Genbank with accession numbers OP223318 for the IST and OP394152 for the ETF-1α region. Blastn results indicated that the ITS and TEF-1α sequences from the isolated strain shared 96.25% and 98.32% similarity with two strains of F. fujikuroi (GenBank Accession No. MF281286.2, and MK311296.1). The isolates were clustered with F. fujikuroi clade (Supplementary Fig.1 and Supplementary Fig.2), consistent with the morphological identification. To evaluate Koch's postulates, 20μL of spore suspension (1×106 spores/mL) were inoculated into ten healthy flat peach fruits with sterile syringes, while another ten healthy fruits were injected with sterilized water as controls. All fruits were kept in sealed plastic boxes at 25 °C with 90% relative humidity. All inoculated fruits showed symptoms similar to those of naturally infected fruits, while uninoculated fruits remained healthy after 4 days. In addition, F. fujikuroi was an important pathogen causing bakanae disease in rice (Hou et al. 2017). This pathogen was also reported to cause Lilium lacifolium Thunb bulb rot (Fang et al. 2022) and cause root rot in peanuts in China (Sun et al. 2022). However, to the best of our knowledge, this is the first record of F. fujikuroi causing fruit rot in flat peach in China. The discovery will provide helpful information about flat peach rot disease management.