First Report of a New Leaf Spot Caused by Plectosphaerella cucumerina on Field Grown Endive (Cichorium endivia) in Italy

During summer 2012, symptoms of a new leaf spot disease were observed in several commercial fields in Treviglio (Bergamo, northern Italy) on plants of curly (Cichorium endivia var. crispum) and Bavarian (C. endivia var. latifolium) endive (Asteraceae). This crop is widely grown in the region for fresh market. The first symptoms on leaves of affected plants consisted of small (1 mm) black-brown spots of irregular shape, later coalescing into larger spots, up to 10 to 15 mm diameter. Eventually, spots were surrounded by a yellow halo. Particularly, affected tissues rotted quickly under high moisture. Disease severity was greatest at 75 to 90% RH and air temperature between 23 and 30°C, where affected tissues rotted quickly. This disease resulted in severe production losses. On one farm in particular, three different fields totaling 2 ha, 5 to 13% of the plants were affected. Diseased tissue was excised, immersed in a solution containing 1% sodium hypochlorite for 60 s, rinsed in water, then placed on potato dextrose agar (PDA) medium, containing 25 mg/liter of streptomycin sulphate. After 5 days, a fungus developed producing a whitish-orange mycelium when incubated under 12 h/day of fluorescent light at 23°C. The isolates obtained were purified on PDA. On this medium, they produced hyaline elliptical and ovoid conidia, rarely septate, measuring 5.0 to 9.0 × 1.7 to 3.9 (average 6.0 × 2.9) μm. Conidia were born on phialides, single, clavate, and 2.8 × 1.4 μm. Such characteristics are typical of Plectosphaerella sp. (1,2). The internal transcribed spacer (ITS) region of rDNA was amplified using the primers ITS1/ITS4 (3) and sequenced. BLAST analysis of the 530-bp segment obtained from C. endivia var. crispum isolate PLC28 and of the 527-bp from C. endivia var. latifolium isolate PLC 30, respectively, showed 99% similarity with the sequence of Plectosphaerella cucumerina (anamorph Plectosporium tabacinum), GenBank EU5945566. The nucleotide sequences of isolates PLC 28 and PLC 30 have been assigned the GenBank accession numbers KC293994 and KC293993, respectively. To confirm pathogenicity, tests were conducted on 30-day-old C. endivia plants. C. endivia var. crispum cv Myrna and C. endivia var. latifolium cv. Sardana plants, grown in 2-liter pots (1 plant per pot, 10 plants per treatment) were inoculated by spraying a 10⁶ CFU/ml conidial suspension of the two isolates of P. cucumerina, prepared from 10-day-old cultures, grown on PDA. Inoculated plants were maintained in a growth chamber at 25 ± 1°C and 90% RH for 5 days. Non-inoculated plants, only sprayed with water, served as controls. All plants inoculated with the two isolates, showed typical leaf spots 7 days after the artificial inoculation, similar to those observed in the field. Later, spots enlarged and leaves rotted. Non-inoculated plants remained healthy. P. cucumerina was reisolated from inoculated plants. The pathogenicity tests were conducted twice with identical results. This is, to our knowledge, the first report of P. cucumerina on endive n Italy, as well as worldwide. Due to the importance of the crop in Italy, this disease can cause serious economic losses.References: (1) A. Carlucci et al. Persoonia 28:34, 2012. (2) M. E. Palm et al. Mycologia 87:397, 1995. (3) T. J. White et al. PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, San Diego, 1990.