In April 2005, an Alternaria sp. was isolated from carrot (Daucus carota) roots harvested in the fall of 2004 and held at 1 to 3°C in a storage facility in Newaygo County, MI. The pathogen was readily isolated on water agar from root tissue exhibiting grayish black, sunken lesions. Morphological characteristics were noted 5 to 7 days after single-conidium cultures were established on potato dextrose agar (3). Sixteen Alternaria sp. isolates were recovered. Cultures were dark olive brown, and conidia were pigmented, ellipsoidal, and produced singly or in chains of two. Conidia were 35 to 45 μm long and 15 to18 μm in diameter, usually with three to eight transverse and one to four longitudinal septa. Pathogenicity of isolates was tested on carrot roots in the laboratory and carrot seedlings (cv. Goliath) in the greenhouse. In the laboratory, four surface-sterilized, whole carrot roots were sprayed until runoff with 2 × 106 conidia/ml of each isolate and incubated at 23 to 25°C in a moist chamber for 10 days. Controls were sprayed with sterile distilled water. Ten to fifteen days after inoculation, inoculated carrots exhibited grayish black, sunken lesions, and an Alternaria sp. was reisolated from the margin of the lesions. Controls remained healthy. In the greenhouse, seven pots containing one 2-week-old carrot seedling were watered to saturation and plants were sprayed until runoff with 2 × 106 conidia/ml for each isolate. Control plants were sprayed with sterile distilled water. After inoculation, plants were enclosed in clear plastic bags, placed under 63% woven shade cloth and watered regularly. Black lesions were observed on the foliage 7 days after inoculation, and wilt and death of plants were observed 15 to 30 days after inoculation. Alternaria sp. was reisolated from the foliage of symptomatic plants. Control plants remained healthy. DNA was extracted from all isolates, and the nuclear ribosomal internal transcribed spacer (ITS) region amplified with primers ITS4 and ITS5 and sequenced. A portion of the ITS sequence has been deposited in the NCBI database (GenBank Accession No. DQ394073). A BLAST search of the NCBI database with the ITS sequences revealed A. radicina, Accession No AY154704, as the closest match with 100% sequence similarity. In September 2005, an Alternaria sp. was isolated from black lesions on carrot roots, crowns, and foliage that were collected from fields in Newaygo and Oceana counties, MI. The recovered isolates were morphologically similar to A. radicina isolates obtained from stored carrots in April 2005. First isolated and identified on stored carrots in New York (3), A. radicina is also present in other carrot-producing areas of the United States (1) and was isolated not only from stored carrots but also from carrots in the field (2) and carrot seeds (4). To our knowledge, this is the first report of A. radicina on stored and field carrots in Michigan, which signifies a serious risk to a carrot industry that ranks among the top five in the United States. References: (1) D. F. Farr et al. Fungi on Plants and Plant Produce in the United States.The American Phytopathological Society, St. Paul, MN, 1989. (2) R. G. Grogan and W. C. Snyder. Phytopathology 42:215, 1952. (3) F. C. Meier and E. D. Eddy. Phytopathology 12:157, 1922. (4) B. M. Pryor and R. L. Gilbertson. Plant Dis. 85:18, 2001.
Geographic Distribution and rDNA-ITS Region Sequence Diversity of Waitea circinata var. circinata Isolated from Annual Bluegrass in the United States
