First report of tomato wilt caused by Fusarium brachygibbosum in China.

Abstract

Tomato (Solanum lycopersicum) is a common vegetable in the Solanaceae family, which as a nutritious vegetable, is widely planted in China. In July 2022, typical wilt symptoms were observed in tomato fields located in the regions of Shiyan (31.343781°N, 110.901005°E), Hubei. Surveys were performed on tomato plants showing symptoms of leaf chlorosis, dry wilt, and vascular wilts on stem and root. The disease incidence ranged from 40% to 70% in 12 surveyed fields, 11.2 ha in total.. After using a sterilized scalpel to cut a small piece of diseased tomato stem and root tissue, diseased tissue was surface disinfested in 75% ethanol for 30s, placed onto potato dextrose agar (PDA) medium,and incubated at 25℃ for 3 days. Then, the emerging single fungal hypha tip was cut and transferred into PDA plates to obtain single spore isolates. Sixteen fungi growing on PDA plates were initially white colonies with abundant aerial mycelium. After seven days of growth, the center of the plate was yellow to orange and red pigmentation was produced. The five-day-old cultures grown on mung bean medium produced macroconidia that were scarce and scattered, three to four septa, wide central cells, slightly sharp apexes, and ranged in size from 12.6-23.6 μm×2.8-4.1 μm (n=30). Microconidia were slightly curved, ovoid with zero to two septa, measuring 5.2-11.8 μm×1.8-2.7μm (n=30). Spherical chlamydospores were terminal or intercalary, and measured a diameter of 8.1-11.6 μm (n =30). Therefore, sixteen isolates were morphologically identified as Fusarium spp. Moreover, the genomic DNA of the isolates (HBSY-1,HBSY-2, and HBSY-3) was extracted for amplification and sequencing of the regions of internal transcribed spacer (ITS) (White et al., 1990), nuclear large subunit rRNA (nLSU) (O'Donnell, 1992; Vilgalys and Hester, 1990), and the translation elongation factor 1-alpha (EF1-α) (O'Donnell et al. 1998) with primers ITS1/ITS4, NL1/LR3, and EF1/2, respectively. Sequences were submitted to GenBank under accession numbers OP959509, OQ568650, OQ568651 (ITS), OQ186731, OQ568652, OQ568653 (nLSU), and OP957576, OQ572485, OQ572486 (EF1-α). BLASTn showed that the sequences ITS, nLSU, and EF1-α were matched 99.61% (508/510 bp; KU528864.1), 99.90% (993/994 bp; GQ505450.1), and 99.85% (651/652; ON032449.1) to Fusarium brachygibbosum, respectively. Multilocus phylogenetic analysis showed that the isolate was of the same clade as F. brachygibbosum. Therefore, morphological characterization and molecular data identified the fungus as F. brachygibbosum. Pathogenicity of the isolate (HBSY-1) was tested on ten seedlings of tomato (cv. Hezuo908). Tomatoes were inoculated by spraying with conidial suspensions (1×107 spores/mL) at the rootstock region of each plant. In addition, ten negative control plants were treated with sterile water. All plants were incubated in an artificial climate box (LongYue, ShangHai) at 25℃ for 12 days. The experiment was repeated three times. Twelve days later, inoculated tomatoes developed typical wilting symptoms of leaves and vascular wilts of stem and root, while the control plants remained healthy. Thus, pathogens were reisolated from the stems of inoculated plants and not from control plants. To our knowledge, this is the first report of F. brachygibbosum causing leaf wilt and vascular wilts of stem and root on tomatoes in China.