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- Cocksfoot mottle virus (CfMV) is a mechanically and beetle-transmitted, non-seedtransmitted
sobemovirus associated with orchardgrass (Dactylis glomerata L.) stand decline
in Europe, Japan, New Zealand (Mahy and Van Regenmortel 2010), and Canada (Bittman
et al. 2006). Additional hosts, reported from New Zealand (Delmiglio et al. 2010), include
Festuca novae-zelandiae, Lolium spp., Poa anceps, Poa cita, Chionochloa rubra, and
Dichelachne crinita. To determine if the virus is present in the United States, surveys for
CfMV were conducted in 2014 and 2015 in orchardgrass seed production fields in the
Willamette Valley in Oregon, where most of the orchardgrass seed produced in the United
States is grown. During June, in each of 2014 and 2015, 18 orchardgrass fields were
selected arbitrarily. Stand age of the seed production fields ranged from 2 to 24 years.
Two of the fields were sampled in both years. Four samples (each containing 4 leaves, one
from each of four plants) were collected along each of four transects in an M-shape pattern
from each field. All plants sampled appeared healthy. In 2014, samples were placed in
separate plastic bags and in 2015 samples were placed in deep well plates (VWR,
International LLC, Radnor, PA). In each year, samples were transported over ice and stored
at 5°C until processed. Samples were homogenized and tested for CfMV using DAS-ELISA,
with antibodies derived from an isolate of CfMV from British Columbia, Canada. In 2014
and 2015, CfMV was detected in 61% and 72% of the fields, respectively. Symptoms were
not present at the time of sampling. In 2015, eight ELISA positive samples were further
examined with RT-PCR. RNA was extracted with the Direct-zol RNA MiniPrep kit (Zymo Research, Irvine, CA) following the manufacturer’s instructions and used for cDNA synthesis
using the SuperScriptIII First-Strand Synthesis SuperMix kit (Life Technologies,
ThermoFisher, Waltham, MA). PrimeSTAR HS DNA Polymerase (Takara Clonetech, Kyoto,
Japan) was used following the manufacturer’s instructions with CfMV specific primers CfCPF1
(GATGGAGCCAGTCTCTCG) and CfCP-R2 (ATCCGTCAATCTTCAAGC) (Delmiglio 2008) for
PCR with the following program: 98°C for 2 min; 40 cycles of 98°C for 15 s, 55°C for 5 s,
and 72°C for 45 s; followed by 72°C for 7 min. The predominant band at 750 bp was
reamplified and sequenced using CfCP-F1 and CfCP-R1 primers. A BLAST search indicated
94.4 to 95.4% identity with CfMV isolates found in native and exotic grasses of New
Zealand (Delmiglio 2008). Sequences were submitted to GenBank (Accession No.
KT984653 to KT984660). To our knowledge, this is the first report of CfMV in orchardgrass
seed production fields in Oregon and in the United States. Results of this study are
significant given the severity of CfMV in Canada and elsewhere, and this could account for
the winter die-out occurring in orchardgrass seed-production fields in Oregon. Additional
studies will be needed to determine to what extent CfMV is associated with stand decline in
Oregon and what vectors may be present.
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- Alderman, S., Martin, R. C., Gilmore, B. S., Martin, R. R., Hoffman, G. D., Sullivan, C. S., & Anderson, N. P. (2016). First Report of Cocksfoot mottle virus Infecting Dactylis glomerata in Oregon and the United States. Plant Disease, 100(5), 1030. doi:10.1094/PDIS-09-15-1017-PDN
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Previous issue date: 2016-05
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