First Report of Fusarium oxysporum f. sp. lactucae Race 1 Causing Fusarium Wilt of Lettuce in Norway

Lettuce (Lactuca sativa L.) is produced in Norway both in field and greenhouses. In Norway, greenhouse lettuce is one of the most important vegetables grown year-round. In winter 2018, wilting symptoms were observed on soil-grown lettuce of the cultivar Frillice in a greenhouse in south east Norway (Buskerud county). Affected plants showed stunted growth, wilting of outer leaves, and brownish discoloration of vascular tissues of taproots and crowns. According to the producer, the disease led to an estimated 10% of yield losses. Fungal isolates were obtained from crowns and roots of diseased plants collected from the greenhouse in 2018 and 2019. Two single spore isolates, 231274 from 2018 and 231725 from 2019, were used in further studies. The isolates were incubated on synthetic nutrient-poor agar (SNA) at 18-20 ⁰C, and a 12 hours dark, 12 hours UV light cycle. Isolate 231274 produced abundant macro- and microconidia characteristics of Fusarium oxysporum while macroconidia were never observed in isolate 231725. On potato dextrose agar (PDA), colonies of isolate 231274 were purple in color and colonies of isolate 231725 were pinkish with abundant aerial mycelium. For PCR-assay, DNA from mycelia was extracted using Easy-DNA kit (Invitrogen). A portion of the translation elongation factor 1-α (EF1-α) gene was amplified using primers F-728F (Carbone and Kohn. 1999) and EF2 (O'Donnell et al. 1998) as described by Aas et al. 2018. Blast analysis of both sequences (accession no. MW316853 for 231274 and MW316854 for 231275) obtained a 99% homology with the sequence of Fusarium oxysporum f.sp. lactucae (FOL) race 1 strain S1 (accession no. DQ837657)(Mbofung et al. 2007). Both isolates were identified as race 1 by using specific primers Hani3' and Hanilatt3rev (Pasquali et al. 2007) as described by Cabral et al. 2014. To complete Koch's postulate, lettuce plants of the cultivar Frillice were used. Race identity was confirmed using the differential lettuce cultivars Costa Rica No.4 (resistant to FOL race 1), Banchu Red Fire (resistant to FOL races 2 and 4) and Romana Romabella (resistant to FOL races 1 and 2) (Gilardi et al. 2017) provided by the breeding company Rijk Zwaan (De Lier, The Netherlands). For inoculation, roots of six 2-weeks old seedlings per cultivar were dipped in a spore suspension (1 x 106 CFU/ml) for 1 min, while controls were dipped in distilled water. Seedlings were planted in 250 ml pots containing fertilized potting substrate, and were placed i