First Report of Sclerotinia sclerotiorum on Calceolaria integrifolia in Italy.

Calceolaria integrifolia L. is an ornamental species grown as a potted plant in Liguria, northern Italy. In the winter of 2006, extensive chlorosis was observed on approximately 10% of the 10-month-old potted plants in a commercial greenhouse. Initial symptoms included stem necrosis and darkening of leaves. As stem and foliar necrosis progressed, infected plants wilted and died. Wilt occurred on young plants within a few days after the initial appearance of symptoms. Infected plants were characterized by the presence of soft, watery tissues that became covered with white mycelium and dark sclerotia. The diseased stem tissue was surface sterilized for 1 min in 1% NaOCl and plated on potato dextrose agar (PDA) amended with 100 mg/liter of streptomycin sulfate. Sclerotinia sclerotiorum (Lib.) de Bary (3) was consistently recovered from infected stem pieces. Sclerotia observed on infected plants measured 0.7 to 1.0 × 2.8 to 4.4 mm (average 1.6 to 2.1 mm). Sclerotia produced on PDA measured 1.0 to 1.1 × 3.0 to 4.2 mm (average 1.7 to 2.3 mm). The internal transcribed spacer (ITS) region of rDNA was amplified with primers ITS4/ITS6 and sequenced. BLASTn analysis (1) of the 522-bp amplicon resulted in 100% homology with the sequence of S. sclerotiorum. The nucleotide sequence has been assigned GenBank Accession No. EU 627004. Pathogenicity of two isolates obtained from infected plants was confirmed by inoculating 10 120-day-old plants grown in individual 14-cm-diameter pots maintained in a greenhouse under partial shade. Inoculum consisted of 1 cm of mycelial plugs excised from a 10-day-old PDA culture of each isolate. Plants were inoculated by placing a mycelial plug on the soil surface around the base of each plant. Ten plants were inoculated per isolate and an equal number of noninoculated plants served as controls. The trial was repeated once. All plants were kept at temperatures ranging between 8 and 17°C (average 12.5°C) and watered as needed. All inoculated plants developed leaf yellowing within 8 days after inoculation, soon followed by the appearance of white mycelium and sclerotia, and then by wilt. Control plants remained symptomless. S. sclerotiorum was reisolated from the stems of inoculated plants. S. sclerotiorum was reported previously on a Calceolaria sp. in the United States (2). To our knowledge, this is the first report of white mold on C. integrifolia in Italy. The economic importance of this disease is currently limited. References (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) Anonymous. USDA Agric. Handb. 165:441, 1960. (3) N. F. Buchwald. Den. Kgl. Veterin.er-og Landbohojskoles Aarsskrift 75, 1949.