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First Report of Turnip mosaic virus in Rhubarb in Alaska. N. L.
Robertson and D. C. Ianson, USDA-ARS, Subarctic Agricultural Research Unit,
Arctic Plant Germplasm Introduction and Research Project, Palmer, Alaska 99645.
Plant Dis. 89:430, 2005; published on-line as DOI: 10.1094/PD-89-0430B. Accepted
for publication 28 December 2004.
In July 2003, noticeable red lesions were observed on rhubarb leaves (Rheum
rhababarum cv. Kerwin) from a plant at the Arctic Plant Germplasm
Research and Introduction Project in Palmer, AK. Extracts of leaf tissue tested
positive for a potyvirus using indirect enzyme-linked immunosorbent assay
(ELISA) and western blots with a monoclonal antibody specific to the potyvirus
group (Agdia, Inc., Elkhart, IN). During the following growing season (June
2004), obvious chlorotic ringspots developed into red lesions on the same
plant and an adjacent plant of the same cultivar. Partially purified particles
that were isolated from the infected rhubarb plants were mechanically inoculated
to an experimental host range (number of infected plants per total number of
plants), resulting in lesions on leaves of Rheum palmatum (1 of 2) and Chenopodium
amaranticolor (3 of 5) but none on C. quinoa
(0 of 4). The leaves with local lesions from C. amaranticolor were ground
in phosphate buffer (1 g of tissue per 10 ml of buffer), and the extract rubbed
onto a set of plants resulting in lesions on R. hybridum (raponticum)
(1 of 2), C. amaranticolor (1 of 4), and C. quinoa (1 of 4). The
original diseased rhubarb plants and experimental symptomatic plants were
confirmed to have a potyvirus using ELISA. Subsequent compound direct ELISA and
western blot assays revealed that the virus reacted strongly to monoclonal or
polyclonal antibodies to Turnip mosaic virus (TuMV) (Agdia, Inc.). Total
RNA was extracted from leaves of the naturally infected rhubarb plants with an
RNeasy Plant Mini Kit (Qiagen Sciences, Germantown, Maryland), and used in
reverse-transcription-polymerase chain reaction (RT-PCR) with specific primers
for TuMV (1) predicted to amplify a 1,134-bp 3(prime)-terminal cDNA fragment
encompassing the 3(prime)-end of the nuclear inclusion protein gene (NIb), the coat
protein gene, and the 3(prime)-nontranslated region. A PCR product of approximately
the expected size was obtained and then sequenced. Sequences (1,077 nt) that
corresponded to the TuMV coat protein gene and 3(prime)-terminal noncoding region
were submitted to Genbank (Accession No. AY744930). Blast searches against NCBI
(National Center for Biotechnology Information) contained high identities to
many TuMV isolates with up to 96% (1,043 of 1,077) nucleotide identity (i.e.,
GenBank Accession No. AF169561). Similar high identities of up to 97% at the
amino acid level occurred within the coat protein coding region (i.e., GenBank
Accession No. BAC02892.1). Infected rhubarb plants were removed from the site
and none of the remaining 109 plants tested positive for TuMV using ELISA. On
the basis of the mechanical transmission to plant hosts, the definitive TuMV
serology, and the consensus of sequenced regions with TuMV, we concluded that
the causal agent of the diseased rhubarb plants was TuMV. Although TuMV has a
wide plant host range occurring worldwide (2), to our knowledge, this is the
first report of TuMV in rhubarb in Alaska and the first time that TuMV has been
detected in Alaska.
References: (1) P. Lehmann et al. Physiol. Mol. Plant Pathol. 51:195,
1997. (2) R. Provvidenti. Page 1340 in: Viruses of Plants. A. A. Brunt et al.,
eds. CAB International, Wallingford, UK, 1996.
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