First Report of Fusarium citri Causing Postharvest Fruit Rot of Chayote in Mexico

Plant Dis. 2022 Nov 19. doi: 10.1094/PDIS-07-22-1554-PDN. Online ahead of print.

Abstract

In November 2018, symptoms of brown rot were observed on chayote (Sechium edule) var. nigrum spinosum with a 20% disease incidence of 120 harvested fruits in the National Germplasm Bank of Sechium edule, located in the Centro Regional Universitario Oriente (CRUO) from the Chapingo Autonomous University (Huatusco, Veracruz, Mexico). For fungal isolation, pieces from symptomatic fruits were surface disinfected by immersion in a 1.5% NaClO solution for 2 min, rinsed in sterile distilled water, placed in Petri plates containing potato dextrose agar (PDA) amended with kanamycin sulfate, and incubated at 25ºC. Fusarium-like colonies were consistently isolated on PDA and five monoconidial isolates were obtained. A representative isolate was selected for morphological characterization, phylogenetic analysis, and pathogenicity tests. On PDA, colonies exhibited white and fluffy aerial mycelia, with diffused pale brown pigment in the center at 7 days of incubation at 25℃ in darkness. Macroconidia (n= 100) were hyaline, falcate, with 4 to 5 septa, measuring 23.9 to 31.9 × 2.9 to 4.2 μm, and foot-shaped basal cells. Microconidia and chlamydospores were absent. Morphological features were consistent with the description of the Fusarium incarnatum-equiseti species complex (Xia et al. 2019). The isolate was deposited as FUS2 in the Culture Collection of Phytopathogenic Fungi of the Laboratory of Plant Pathology at the Colegio de Postgraduados. For molecular identification, genomic DNA was extracted, and the internal transcribed spacer (ITS) region, partial sequences of translation elongation factor 1-alpha (EF1-α), and the second-largest subunit of RNA polymerase II (rpb2) genes were amplified, and sequenced with the primer sets ITS5/ITS4 (White et al. 1990), EF1-728F/EF1-986R (Carbone and Kohn 1999), and RBP2-5F/RPB2-7R (Liu et al. 1999), respectively. DNA sequences were edited in BioEdit 7.2 and compared with those in the NCBI nucleotide database. Alignments were implemented in MEGA X using reference sequences from Fusarium spp. A phylogenetic tree, including published ITS, EF1-α, and rpb2 sequence data, was constructed for the Fusarium incarnatum-equiseti species complex (FIESC) based on Maximum Likelihood. The sequences were deposited in GenBank (accession nos. ON878083, ON890421, and ON890420). The phylogenetic analysis grouped the isolate FUS2 within the F. citri clade. Pathogenicity of the fungus was verified on 10 healthy chayote fruits var. nigrum spinosum previously disinfested by immersion in a 1% NaClO solution for 3 min and washed in sterile water. A total of 5 mL of a conidial suspension (1 × 106 spores/ml) was sprayed on each whole fruit. Ten control fruit were sprayed with sterile distilled water. The fruits were kept in a moist plastic chamber at 25°C and 12 h light/dark for 30 days. All inoculated fruits developed water-soaked brown lesions (3 to 5 cm in diameter) covered with white mycelium at 15 days after inoculation, whereas no symptoms were observed on the control fruits. The fungus was consistently re-isolated only from the diseased fruits and found to be morphologically identical to the isolate used for inoculation, fulfilling Koch´s postulates. Fusarium citri has been associated with Capsicum sp. and mandarin orange in China, Triticum sp. in Iran, alfalfa in Denmark, and lettuce in the Czech Republic and Italy (Farr and Rossman 2022). To our knowledge, this is the first report of F. citri causing postharvest fruit rot of chayote in Mexico and worldwide.

Keywords: Causal Agent; Crop Type; Etiology; Fungi; Subject Areas; Vegetables.