First Report of Black Rot Caused by Phomopsis cucurbitae on Cantaloupe (Cucumis melo) in the Piedmont Region of Northern Italy.

Cucumis melo L., belonging to the Cucurbitaceae family, is cultivated on more than 23,000 ha in Italy. Cantaloupe (C. melo L. var. cantalupensis Naudin) is the most popular variety. In summer 2010, a previously unknown rot was observed on fruits produced in Italy and marketed in the Piedmont Region of northern Italy. Early symptoms on fruit consisted of irregular, brown, soft, sunken lesions up to 10 cm in diameter. No surface mold was visible and pycnidia were not present. Internally, the decay is adjacent to the sunken area of the fruit's surface and is soft, water soaked, spongy, with a nearly circular margin, and easily separated from healthy tissues. Fragments (approximately 3 mm) were taken from the margin of the internal diseased tissues, cultured on potato dextrose agar (PDA), and incubated at 24 ± 1°C, (16 h of light and 8 h of darkness). Fungal colonies initially appeared coarse, at first whitish then buff brown, and produced dark pycnidia 0.5 mm in diameter, which exuded numerous conidia belonging to two types. Type A conidia were hyaline, unicellular, ellipsoidal to fusiform, sometimes slightly constricted in the middle, and measured 5.6 to 10.3 × 1.7 to 2.6 (average 8.0 × 2.1) μm. Type B conidia were hyaline, long, slender, curved, and measured 17.1 to 26.6 × 0.7 to 1.4 (average 22.0 × 1.0) μm. Sclerotia were not produced. The morphological characteristics of the fungus corresponded to those of the genus Phomopsis (1). The internal transcribed spacer (ITS) region of rDNA was amplified using the primers ITS1/ITS4 and sequenced. BLAST analysis of the 543-bp segment showed a 99% similarity with the sequence of a Phomopsis sp. (GenBank Accession No. HM999947). The nucleotide sequence has been assigned the GenBank Accession No. JN032733. Both Phomopsis cucurbitae and P. sclerotioides are pathogenic to Cucurbitaceae, however P. cucurbitae is identifiable by the production of B conidia and the absence of sclerotia. Therefore, P. cucurbitae has been considered the causal agent of the disease. Pathogenicity tests were performed by inoculating three wounded cantaloupe fruits after surface disinfesting in 1% sodium hypochlorite. Six wounds per fruit, 1 cm deep, were made with a sterile needle. Mycelial disks (10 mm in diameter), obtained from PDA cultures of one strain, were placed on each wound. Three control fruits were inoculated with PDA. Fruits were incubated at 16 ± 1°C in the dark. The first symptoms developed 4 days after the artificial inoculation. Two days later, the rot developed at all inoculation points and the pathogen was consistently reisolated. Noninoculated fruit remained healthy. The pathogenicity test was performed twice with similar results. P. cucurbitae has been reported on melon in many countries (2,3). To our knowledge, this is the first report of the disease in Italy. Currently, the relevance of the disease in the country is not yet well known. However, attention must be paid considering that the pathogen can be transmitted through seeds. References: (1) H. L. Barnett and B. B. Hunter. Illustrated Genera of Imperfect Fungi. Burgess Publishing Company, Minneapolis, MN, 1972. (2) L. Beraha and M. J. O'Brien. Phytopathol. Z. 94:199, 1979. (3) E. Punithalingam and P. Holliday. Phomopsis cucurbitae. IMI Descriptions of Fungi and Bacteria. 47, Sheet 469, 1975.