First Report of Leaf Spot Caused by Phoma exigua on Hydrangea macrophylla in Italy.

Hydrangea macrophylla is grown in Italy as a potted plant and also for landscaping. During the fall of 2005 in a nursery located in Lazio (central Italy), a severe foliar disease was observed on 12-month-old potted plants of cv. Hanabi. Small necrotic spots surrounded by chlorotic haloes were observed on the upper side of infected leaves. At temperatures near 20°C and relative humidity ranging between 80 to 90%, spots enlarged to form round areas 2 to 7 cm in diameter that were well defined by a brown margin. Severely infected leaves became chlorotic and abscised. Heavily affected plants were defoliated. Infected plants rarely died, but the presence of lesions on mature plants decreased aesthetic quality and subsequent market value. The disease occurred on 30% of the plants in one nursery. Stems and flowers were not affected by the disease. From infected leaves, a fungus was consistently isolated on potato dextrose agar with 25 mg/liter of streptomycin added. The fungus was grown on leaf extract agar, 30 g of leaves per liter, and maintained at 22°C (12 h of light and 12 h of dark). After 30 days, black pycnidia 275 to 255 μm in diameter developed, releasing conidia that were hyaline, elliptical, nonseptate, and measuring 4.6 to 7.6 (average 6.0) × 1.4 to 4.2 (average 2.6) μm. On oatmeal agar, the addition of a drop of concentrated NaOH caused a positive reaction, turning the medium red (2). On the basis of its morphological characteristics, the fungus was identified as a Phoma sp. The ITS (internal transcribed spacer) region of rDNA was amplified using the primers ITS4/ITS6 (3) and sequenced. BLASTn analysis (1) of the 560 bp obtained showing an E-value of 0.0 with Phoma exigua. The nucleotide sequence has been assigned GenBank Accession No. DQ384612. Pathogenicity tests were performed by spraying leaves of healthy 6-month-old potted H. macrophylla (cvs. Hanabi and Zaffiro) plants with a spore and mycelial suspension (10⁵ CFU/ml). Plants without inoculation served as controls. Five plants were used for each treatment. Plants were covered with plastic bags for 5 days after inoculation and kept in a growth chamber at 20°C with relative humidity at 80 to 90%. The first lesions developed on leaves of cv. Hanabi 12 days after inoculation, while control plants remained healthy. Lesions did not develop on inoculated cv. Zaffiro plants. The fungus was consistently reisolated from the lesions of cv. Hanabi. The pathogenicity test was carried out twice. The presence of P. exigua on H. macrophylla has been reported in the United States (4). In Italy, the disease can be found in a limited area. References: (1) S. F. Altschud et al. Nucleic Acids Res. 25:3389, 1997. (2) G. H. Boerema and L. H. Howeler. Persoonia 5:15, 1967. (3) D. E. L. Cooke and J. M. Duncan. Mycol. Res. 101:667, 1997. (4) M. L. Daughtrey et al. Page 26 in: Compendium of Flowering Potted Plant Diseases. The American Phytopathological Society. St. Paul, MN, 1995.