First Report of Leaf Spot of Milky Bellflower (Campanula lactiflora) Caused by a Phoma sp. in Italy.

Campanula lactiflora (milky bellflower), a perennial herbaceous plant in the Campanulaceae, is used in park and gardens and sometimes cultivated for cut flower production. In June 2008, a previously unknown leaf spot was observed on C. lactiflora 'New Hybrids' plants from an experimental nursery located near Carmagnola (Torino, northern Italy). Leaves of infected plants showed extensive and irregular, dark brown, necrotic lesions that were slightly sunken with well-defined borders. Lesions initially ranged from 0.5 to 3 mm, eventually coalesced, and covered the entire leaf. Black pycnidia (107 to 116 μm in diameter) containing hyaline, ellipsoid, nonseptate conidia measuring 3.7 to 4.7 × 1.2 to 2.0 (average 4.3 × 1.6) μm were observed. On the basis of these morphological characteristics, the fungal causal agent of the disease could be related to the genus Phoma. In some cases, the basal leaves turned completely necrotic and the plant died. The disease affected 50% of plants. Diseased tissue was excised, immersed in a solution containing 1% sodium hypochlorite for 2 to 3 s, rinsed in water, and then cultured on potato dextrose agar (PDA) medium. A fungus developed that produced a greenish gray mycelium with a white border when incubated under 12 h/day of fluorescent light at 22 to 25°C. The internal transcribed spacer (ITS) region of rDNA was amplified using the primers ITS4/ITS6 and sequenced. BLAST analysis (1) of the 459-bp segment showed a 100% similarity with the sequence of a Didymella sp. (synonym Mycosphaerella), anamorphic stage of Phoma spp. The nucleotide sequence has been assigned GenBank Accession No. GU128503. Pathogenicity tests were performed by placing 8-mm-diameter mycelial disks removed from PDA cultures of the fungus isolated from infected plants on leaves of healthy potted 4-month-old C. lactiflora 'New Hybrids' plants. Eight disks were placed on each plant. Plants inoculated with PDA alone served as controls. Six plants per treatment were used. Plants were covered with plastic bags for 4 days after inoculation and maintained in a growth chamber with daily average temperatures ranging between 23 and 24°C. The first foliar lesions developed on leaves 5 days after inoculation, and after 8 days, 80% of leaves were severely infected. Control plants remained healthy. A Didymella sp. was consistently reisolated from leaf lesions. The pathogenicity test was completed twice. To our knowledge, this is the first report of the presence of a Didymella sp. on C. lactiflora in Italy. Mycosphaerella campanulae and M. minor were reported on C. americana and C. lasiocarpa in the United States (2). The economic importance of the disease currently is limited, but could become a more significant problem in the future if the cultivation of this species becomes more widespread. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) D. F. Farr et al. Fungi on Plants and Plant Products in the United States. The American Phytopathological Society, St. Paul, MN, 1989.