First report of postharvest fruit rot of stored apple caused by Neopestalotiopsis clavispora in Serbia

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Publication Details

Journal
Plant Disease
Published
2026-09-20
DOI
https://doi.org/10.1094/pdis-06-26-1092-pdn
Primary Topic
Fungal Plant Pathogen Control
Type
article
Field-Weighted Citation Impact
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article

First report of postharvest fruit rot of stored apple caused by Neopestalotiopsis clavispora in Serbia

Milica Mihajlović, Mila Grahovac, Mladen Petreš, Jelena Vukotić et al.
Plant Disease
Fungal Plant Pathogen Control
article

First report of postharvest fruit rot of stored apple caused by Neopestalotiopsis clavispora in Serbia

Milica Mihajlović, Mila Grahovac, Mladen Petreš, Jelena Vukotić, Dragana Budakov, Marta Loc, Dobrila Radić, Snežana Rajkov, Tatjana Konja
article en

Abstract

Fungal postharvest diseases of apple (Malus domestica Borkh.) cause significant losses during the storage period. Apple fruits (cv. Fuji) with necrotic lesions were collected in late November 2022 from an ultra-low-oxygen cold storage facility in Titel, Serbia, originating from the orchard in Titel (45°12'47.1"N, 20°15'23.6"E). Of 100 fruits examined, 28 (28%) showed symptoms of postharvest rot. Neopestalotiopsis sp. was recovered from two fruits (7.1% of diseased fruits; 2% of all fruits). These fruits exhibited dark brown necrotic lesions, circular and slightly sunken. In advanced stages, the affected tissue collapsed, and small, dark fruiting bodies (acervuli) were observed on the surface of necrotic lesions. Pathogen isolation was performed using standard techniques in plant pathology. Surface-sterilized symptomatic fruits were rinsed with sterile water and aseptically cut in half. Fragments from the margin between healthy and diseased tissue were transferred onto Potato Dextrose Agar (PDA) and incubated at 25±1°C in the dark for seven days. Two isolates formed aerial, white and cottony mycelia. The black acervuli formed on mycelia in concentric circles. Basal and apical cells were hyaline, while other cells were brown to pale brown in color. Conidia were five-celled, fusiform to ellipsoid, with three to four apical appendages. Based on morphological properties, these isolates were identified as Neopestalotiopsis sp., consistent with the descriptions by Maharachchikumbura et al. (2014) and Santos et al. (2019). Pathogenicity was confirmed on apple fruits (cv. Fuji). Three surface-sterilized fruits per isolate were sprayed with 5 ml of conidial suspension (1×10 5 conidia/ml) prepared from Neopestalotiopsis sp. cultured on PDA and incubated at room temperature for 21 days. Inoculated fruits developed the same symptoms as fruits collected from cold storage and reisolated colonies showed morphology identical to that of the colonies used for inoculation. Control fruits treated with sterile water developed no symptoms. The identification was confirmed by amplifying and sequencing three regions using the following primer pairs: Bt2a/Bt2b for β-tubulin region (Nasri et al., 2015), ITS1/ITS4 for ITS region (White et al., 1990) and EF1-728/EF2 for TEF-1α region (O’Donnell et al., 1998; Carbone and Kohn, 1999). Amplicons of expected sizes (500 bp for both Bt2a/Bt2b and EF1-728/EF2 primer pairs and 600 bp for ITS1/ITS4 primer pair) were confirmed by gel electrophoresis and Sanger sequenced (Macrogen Europe BV) in both directions. BLASTn analysis showed that Z23K and Z4K shared a high identity with N. clavispora sequences deposited in GenBank: 99.74 and 99.75% with MZ514840.1, respectively, 99.23 and 99.26% with KU096880.1, respectively, both 100% with LC209221.1 for the β-tubulin region; both 100% for MZ494651.1, KU096881.1 and LC209220.1 for the TEF-1α region; 99.8 and 99.4% with MZ441062.1, 99.79 and 99.58% with KU096879.1 and 99.78 and 99.59% with LC209216.1, respectively, for the ITS region. The sequences were deposited in GenBank (Accession Nos. PZ097812 and PZ097813 for the β-tubulin region, PZ097814 and PZ097815 for the TEF-1α region, OR759142 and OR759140 for the ITS region). To our knowledge, this is the first report of Neopestalotiopsis clavispora causing postharvest fruit rot of apple in Serbia. Together with our recent report of Stemphylium vesicarium (Petreš et al., 2024), these findings highlight the increasing diversity of fungal pathogens associated with stored apple fruits in Serbia.

Plant Disease
University of Novi Sad (RS), Institute of Pesticides and Environmental Protection (RS)
Openalex Percentile: Top 8%
Fungal Plant Pathogen Control
DOISourceOpenAlex
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