First Report of Strawberry Wilt Caused by Fusarium oxysporum Schltdl. in Jordan.

Abstract

Strawberries (Fragaria ananassa Duch.) are grown in Jordan year round due to the diversity of climatic conditions and the possibility of growing local or foreign varieties. More than six thousand greenhouses are planted with strawberries in the highlands and the Jordan Valley. About 12 thousand tons of strawberry are produced annually and 2000 tons are exported to European and Arab countries. In April and May 2019, symptoms of wiltand whole plant collapse were observed on approximately 30% of commercial strawberry cv. Deli gent crop in the Jordan Valley (Dayr Alla long.35.6188766, lat. 32.227465). Plants were either dead or showing symptoms including vascular wilt, external and internal discoloration of the stems, and dead shoots. Forty symptomatic plants were collected from 10 greenhouses, and stem fragments were surface sterilized and plated on potato dextrose agar (PDA). Six fungal isolates showed morphological characteristic of Fusarium oxysporum. Colonies on PDA were purple-violet, floccose, with abundant aerial mycelium; colony margins were irregular. Macroconidia were falcate, apical cells had a blunt or papillate shape, basal cells were foot shaped, three- to five-septate, hyaline, smooth, thin-walled, and 37 - 42 × 3 - 6.0 μm in diameter. Aerial microconidia were abundant, hyaline, ellipsoidal, zero to one-septate: 5 - 11 × 2 - 4.0 μm. Chlamydospores were globose to subglobose, intercalary or terminal, with an average diameter of 12 μm (Figure 1: A, B and C) (Nelson et al, 1983; Leslie and Summerell, 2006). Four representative isolates (FoSB2021JO-02, FoSB2021JO-06, FoSB2021JO-08 and FoSB2021JO-09) were DNA extracted, amplified with the translation elongation factor 1-α (EF1α) gene (EF1/EF2) primers (Geiser et al., 2004), and sequenced at Macrogen Inc, South Korea. Forward and reverse sequences were received, assembled and consensus sequence were produced using the BioEdit sequence alignment editor. Consensus sequences of the four isolates were used to conduct BLASTn queries of NCBI website (https://www.ncbi.nlm.nih.gov) and were 100%, 99.9%, 99.6%, 99.9% identical to F. oxysporum accessions MN417194.1, MK968948.1,MK968948.1, and MK968952.1, respectively. A phylogenetic tree with 1000 bootstraps was created using MEGA 7 software (Kumar et al. 2016) (Figure 2). Similarly, the four isolates were 99.5%, 100%, 100%, and 100% identical F. oxysporum reference accessions AF008507, FJ985275, FJ985275, and FJ985278 in the Fusarium MLST database, respectively. Consensus sequences of the four isolates were submitted to GenBank and accession numbers were assigned (OK040155 - OK040158). For pathogenicity tests on strawberries, a spore suspension of 1 × 105 conidia/ml was prepared separately for six isolates. Roots of identical 2-month-old healthy strawberry seedlings (15 plants of cv. Deli-gent) were cut and dipped in the spore suspensions for 30 min. They were then planted in 25 x 20 cm deep plastic pots filled with a sterile mixture of peat - moss, perlite, and vermiculite (60:20:20v/v). Control strawberry plants were soaked in water prior to planting. All plants were placed in a greenhouse at 25°C ±2 along with 15 uninoculated control plants. After 30 days, inoculated plants displayed similar symptoms to those observed in the green house, whereas control plants were symptomless. Roots from symptomatic plants were cultured on PDA and F. oxysporum was recovered and identified morphologically as F. oxysporum. To our knowledge, this is the first report of Fusarium wilt of strawberry in Jordan. The pathogen can cause significant economic losses to strawberries in Jordan and worldwide. Therefore, it is extremely important for disease control in nurseries to determine the infection source and possible factors that increase the incidence of infection to control the disease.