First Report of Brown Root Rot of Alfalfa Caused by Phoma sclerotioides in Colorado and New Mexico.

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 winterkill, slow emergence from winter dormancy, and yield loss of alfalfa (Medicago sativa L.) (1,2). BRR is a problem in regions with severe winters and is common in Alaska and Alberta, Saskatchewan and Manitoba, Canada. It was first observed in the continental United States in Wyoming during 1996 (2) and has subsequently been found in Idaho, Minnesota, Montana, New Hampshire, New York, Vermont, and Wisconsin. In the intermountain valleys of northern New Mexico and western Colorado, winters can be severe; alfalfa winterkill events occur periodically, but it is unknown if BRR is present. In May 2006, alfalfa plants were collected from production fields in Huerfano, Otero, and Rio Grande counties in Colorado and Rio Arriba and Taos counties in New Mexico and assessed for BRR. Two to three fields were sampled per county and 20 or 40 plants were collected per field. All fields existed for at least two winters. Fields sampled in Rio Grande County exhibited severe winterkill, with most plants completely girdled by crown lesions. Plants from other fields exhibited a range of root and crown rots. Isolation of P. sclerotioides was attempted from all plants with a previously described protocol (4). The pathogen was isolated from crown lesions of one alfalfa plant each from Rio Grande and Taos counties. Both lesions extended into the cortex. On potato dextrose agar and water agar with barley (4), single-conidium cultures of each isolate produced large pycnidia (0.35 to 0.80 mm in diameter) with multiple beaks, white cirri darkening to yellow with age, and unicellular, hyaline, ovoid conidia 5 to 7 μm long by 2 μm wide. Diagnostic PCR of the cultures using P. sclerotioides-specific primers (3) resulted in a single amplicon of expected size (500 bp). The internal transcribed spacer (ITS) 1, 5.8S, and ITS2 of the rDNA were amplified and sequenced using primers ITS1 and ITS4. The ITS sequences (GenBank Accession Nos. EU265669 and EU265670) were >98% identical to P. sclerotioides ATCC isolate 56515 over 503 bp of aligned sequence. Potted 'Vernal' alfalfa was inoculated 4 months after seeding, kept at 4°C for 5.5 weeks, 0 to -2°C for 12 weeks, and 4°C for 3 weeks. Of the 14 plants inoculated with the Colorado isolate, 11 developed cortical lesions and 8 winterkilled. Of the 23 plants inoculated with the New Mexico isolate, 22 developed cortical lesions and 16 winterkilled. Lesions were light to very dark brown, sometimes with a darker border and often containing abundant pycnidia. Winterkill was associated with lesions girdling the crown. P. sclerotioides was isolated from the lesions. To our knowledge, this is the southernmost report of BRR in North America and the first report of BRR in New Mexico and Colorado. The incidence and severity of BRR in the region surveyed appear to be considerably lower than in the more northern regions.