In the UK, lettuce is produced both in the field and in greenhouses or polytunnels. In summer 2022, wilt symptoms were observed for the first time on lettuce (cv. Amica) grown in soil in part of a single 0.55 ha greenhouse in County Armagh, Northern Ireland (NI). Initial presentation of symptoms was stunting of plants, followed by wilting and yellowing of lower leaves in approx. 12% of the plants. Orange-brown discoloration of vascular tissue in the tap root of affected plants was also observed. To isolate the causal pathogen, sections (0.5 cm2) of symptomatic vascular tissue from 5 plants were surface sterilized in 70% ethanol for 45 s, washed twice in sterile water and placed on potato dextrose agar (PDA) amended with 20 µg mL-1 chlortetracycline. Plates were incubated at 20°C for 5 days and fungal colonies sub cultured onto PDA. Isolates from all five samples displayed morphology typical of Fusarium oxysporum and were cream to purple in colour with abundant microconidia and occasional macroconidia. DNA was extracted from 5 isolates and part of the translation elongation factor 1- α (EF1-α) gene amplified by PCR and sequenced as described by Taylor et al. (2016). All EF1-α sequences were identical (OQ241898) and matched with F. oxysporum f. sp. lactucae race 1 (MW316853.1, isolate 231274) and race 4 (MK059958.1, isolate IRE1) with 100% sequence identity using BLAST. Isolates were then identified as FOL race 1 (FOL1) using a race-specific PCR assay (Pasquali et al. 2007). Next, pathogenicity and race identity of one isolate (AJ773) was confirmed using a set of differential lettuce cultivars (Gilardi et al. 2017); Costa Rica No. 4 (CR; FOL1 resistant), Banchu Red Fire (BRF; FOL4 resistant) and Gisela (GI; FOL1 / FOL4 susceptible). Plants were inoculated with AJ773 (this study), ATCCMya-3040 (FOL1, Italy; Gilardi et al. 2017) and LANCS1 (FOL4, UK; Taylor et al., 2019). Roots of 16-day-old lettuce plants (8 replicates per cultivar/isolate) were trimmed and soaked in a spore suspension (1 x 106 conidia mL-1) for 10 min before transplanting into compost in 9 cm pots. Control plants for each cultivar were dipped in sterile water. Pots were placed in a glasshouse (25C day, 18C night). Inoculation with AJ773 and FOL1 ATCCMya-3040 resulted in typical symptoms of Fusarium wilt for BRF and GI 12-15 days after inoculation, while for FOL4 LANCS1, wilting was observed in CR and GI. Thirty-two days after inoculation, plants were cut longitudinally and vascular browning was observed in all plants where wilt was present. All uninoculated control plants, CR inoculated with FOL1 ATCCMya-3040 or AJ773, and BRF inoculated with FOL4 LANCS1 remained healthy. These results confirm the identity of isolate AJ773 from NI as FOL1. Koch's postulates were fulfilled by consistent re-isolation of F. oxysporum from BRF and GI plants and identifying as FOL1 using the race-specific PCR. No FOL was re-isolated from control plants of any cultivar. Fusarium wilt in England and the Republic of Ireland first reported by Taylor et al. (2019) was identified as FOL4 and has been confined to indoor lettuce production with further outbreaks caused by the same race. However, FOL1 was recently identified in Norway in a soil-grown glasshouse crop (Herrero et al. 2021). The presence of both FOL1 and FOL4 in neighbouring countries within the UK is a serious risk to lettuce production and is of particular importance to growers who rely on knowledge of cultivar resistance to specific FOL races to make decisions on which variety to plant.
Keywords: Causal Agent; Crop Type; Fungi; Pathogen detection; Subject Areas; Vegetables.