First Report of Fusarium oxysporum f. sp. fragariae Causing Fusarium Wilt on Strawberry (Fragaria × ananassa) in Saudi Arabia.

Abstract

Recently, Saudi growers have expanded their production of organic, soilless-grown strawberries (Fragaria × ananassa Duch.), but their production shows many difficulties associated with disease susceptibility. In October 2021, 45% of strawberry plants cv. "Festival" organically cultivated in Dammam city, Saudi Arabia (26°31'34.5"N 50°00'51.0"E) showed wilting symptoms. Typical symptoms were yellowing, rapid wilting, death of older leaves, stunting, and decreased roots. Vascular bundle necrosis and crown and root rot were also observed; plants eventually collapsed and died. Twenty symptomatic strawberry plants were sampled to isolate the pathogen. Pieces (4 × 4 mm) of the symptomatic tissues from crowns and roots were sanitized with 1% NaOCl (90 s), submerged in 70% alcohol (20 s), rinsed with sterile water (2x 30 s), placed on potato dextrose agar (PDA; Scharlau Chemie, Spain) and incubated at 25°C for 6 days. Next, we prepared single-spore cultures on PDA and synthetic nutrient-poor agar (SNA). On PDA media, pure cultures produced abundant aerial mycelium, with light pink or purple pigmentation in the medium after incubation at 25°C for 7 days. On SNA media, aerial microconidia were abundant cylindrical to ellipsoid hyaline with zero to one septate (3.8 - 5.9 × 1.3 - 2.5 μm, n = 50). Macroconidia were few, hyaline and falcate, with slightly curved apexes and 2 to 4 septate (18.9 - 27.5 × 3.3 - 4.6 μm, n = 50). Chlamydospores were roundish and terminal or intercalary. As Leslie and Summerell (2006) described, such morphological characteristics are typical of F. oxysporum. The isolates' identities were established by extracting DNA using the DNeasy Plant Mini kit (QIAGEN, Hilden, Germany). This was followed by amplification and sequencing of the internal transcribed spacer (ITS) (White et al., 1990), elongation factor 1-α (TEF1-α) (O'Donnell et al., 1998), and the ribosomal RNA intergenic spacer (IGS) (Canizares et al., 2015). The ITS, TEF1-α, and IGS sequences of an isolate Fof-10 were submitted to GenBank (PP564462, PP703242, and PP784894, respectively). BLAST analysis confirmed 99.71 and 100% identities to the ITS, TEF-1α, and IGS sequences of F. oxysporum (KU931552.1, OR640020.1, and FJ985519.1), respectively. All isolates tested were confirmed at the forma specialis fragariae, level using the specific primers FofraF/FofraR (Suga et al. 2013). The ∼239 bp amplicon was sequenced and submitted to GenBank (PP703243). Two-month-old healthy strawberry plants of cultivars "Festival," "Marquis," and "Monterey" were inoculated by dipping the roots in the spore suspension (107 conidia ml-1) for 15 min (Henry et al. 2017). There were five replicates for each cultivar. Plants dipped in water were used as a control treatment. The plants were transplanted in sterilized soil and placed in a greenhouse at 30/26°C (day/night). Within 4 to 6 weeks, inoculated plants showed severe wilting and discoloration of the internal crown tissue, while control plants were symptomless. The pathogen was re-isolated from the discolored vascular tissue onto PDA and identified morphologically and molecularly as the original one, thus fulfilling Koch's postulates. The test was repeated twice. This report confirms F. oxysporum f. sp. fragariae as a causal agent of Fusarium wilt of strawberries in Saudi Arabia. This pathogen was previously reported to cause the Fusarium wilt of strawberries in California (Dilla-Ermita et al., 2023). This disease has been observed in several hydroponic strawberry greenhouses in Saudi Arabia, with incidence ranging from 25% to 45% across multiple locations. Given this, proper strategies are needed to manage this disease and to be compatible with organic farming.