First Report of Brown Root Rot of Alfalfa Caused by Phoma sclerotioides in Maine, Ontario, and Pennsylvania.

Abstract

Brown root rot (BRR), caused by the fungal pathogen Phoma sclerotioides G. Preuss ex Sacc. (synonym Plenodomus meliloti Dearn. & G.B. Sanford), is associated with yield loss of alfalfa (Medicago sativa L.) in regions with severe winters (1). In the spring of 2007, 9 to 69 alfalfa plants were collected from each of five production fields in Maine, 10 fields in Ontario, and nine fields in Pennsylvania. All alfalfa stands existed at least two winters. P. sclerotioides was isolated from alfalfa root and crown lesions from five fields in Maine (Penobscot, Somerset and Waldo counties), seven fields from southwestern (Woodstock and Niagara), south-central (Lindsay and Belleville), and southeastern Ontario (near Ottawa), and four fields in Pennsylvania (Columbia, Crawford, and Jefferson counties; 41.1 to 41.6°N). BRR incidence was 9 to 29% in Maine, 5 to 29% in Ontario, and 8 to 22% in Pennsylvania. In Ontario, some lesions girdled the crown; in three fields in Maine, large pycnidia characteristic of P. sclerotioides were present on alfalfa crowns and overwintered stems. On potato dextrose agar, conidia (5 to 8 × 2 to 3 μm, unicellular, hyaline, and ovoid) and pycnidia (0.33 to 1.15 mm in diameter with multiple beaks) of single-conidium isolates were characteristic of P. sclerotioides (2). Diagnostic PCR (3) of isolates resulted in a single amplicon of expected size (500 bp). The internal transcribed spacer (ITS) 1, 5.8S, and ITS2 of the rDNA were sequenced for 12 representative isolates, and sequences (GenBank Accession Nos. FJ179151 to FJ179162) were 95.5 to 100% identical to P. sclerotioides ATCC isolate 56515 over a 488-bp alignment. Eight months after seeding, potted 'Vernal' alfalfa was inoculated (4), kept at 4°C for 8 weeks, 0 to -2°C for 12 weeks, 4°C for 8 weeks, and 10 to 15°C for 7 weeks. Of 108 plants inoculated with the Maine isolates, 35 developed severe cortical lesions and 16 died. Of 18 plants inoculated with the Ontario isolates, 16 developed severe cortical lesions and eight died. Of 18 plants inoculated with a Pennsylvania isolate, 11 developed severe cortical lesions and five died. Lesions were typical of BRR: light to very dark brown, sometimes with a darker border, and often containing abundant pycnidia. Plant mortality was associated with lesions that girdled the root and crown. Of 18 plants in the control treatment, three developed severe cortical lesions and none died. BRR is common in Alberta, Saskatchewan, and Manitoba, but in eastern Canada it has been reported only in Nova Scotia. To our knowledge, this is the first report of BRR in Maine, Ontario, and Pennsylvania and the southernmost report of BRR in eastern North America.