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Plant Health Brief
First Report of Lilac ring mottle virus Infecting

Lilac in the United States
Dipak Sharma-Poudyal and Nancy K. Osterbauer, Oregon Department of Agriculture, Salem 97301; Melodie L. Putnam, Oregon State
University Plant Clinic, Corvallis 97331; and Simon W. Scott, Clemson University, Clemson, SC 29634
Accepted for publication 26 May 2016. Published 14 July 2016.
Sharma-Poudyal, D., Osterbauer, N. K., Putnam, M. L., and Scott, S. W. 2016. First report of Lilac ring mottle virus infecting lilac in the United States.
Plant Health Prog. 17:158-159.
In May 2015, the Oregon Department of Agriculture investi-

gated a report of potential virus symptoms being observed on lilac
plants (Syringa vulgaris L.) produced by a nursery in Marion
County. Simultaneously, symptomatic lilac plant samples were
received by the Oregon State University Plant Clinic from a
customer of the same nursery. Lilac ‘President Grevy’ plants
inspected at the nursery showed symptoms of leaf deformation,
reduction in leaf size, ring spots, and line patterns (Fig. 1). Two
months later, similar foliar symptoms were observed on lilac
‘Krasavitsa Moskvy’ plants at the same nursery (Fig. 2). Symp-
toms on both cultivars resembled those reported for Lilac ring
mottle virus (LiRMoV) (Van Der Meer et al. 1976). LiRMoV is
an isometric RNA virus within the family Bromoviridae and
genus Ilarvirus (Scott and Ge 1995; Scott and Zimmerman 2008)
and has only been reported from the Netherlands. The virus was
sap transmissible to herbaceous hosts under experimental
conditions and was seed transmissible in the experimental hosts
Chenopodium quinoa, C. amaranticolor, and Celosia argentea
(Van der Meer et al. 1976). Symptom expression in infected lilac
is influenced by environmental conditions and may be erratic;
thus, infections may remain cryptic for years (Van der Meer et al.
1976). FIGURE 1
To determine if LiRMoV was present in the symptomatic lilacs,
Foliar symptoms caused by Lilac ring mottle virus on lilac ‘President
total nucleic acids (TNA) were extracted from symptomatic leaf
Grevy.’
tissues using a procedure modified from Hughes and Galau
(1988). The TNA was used in ONE-STEP PCR reactions
(Qiagen, Germantown, MD) at a melting temperature of 55°C and
with primers (downstream: 5′-GAGACCGAAGTCTTCTTCC-3′
and upstream: 5′-CCACGTGCTTCTCACCC-3′) specific for the
movement protein of the RNA3 of LiRMoV (GenBank Accession
No. U17391) (Scott and Zimmerman 2008). In addition to the
TNA from the samples, a positive control (plasmid pLRMV-7)
(Scott and Zimmerman 2008) and negative controls (healthy plant
tissue and water) were analyzed in concurrent PCR reactions. The
anticipated 649-bp amplicon was produced in the lilac samples
and in the positive control, but not in the negative controls. The
amplicons from seven lilac samples were cloned using the pGem-
T Easy Vector (Promega, Madison, WI), selected by blue-white
screening, and then sequenced using the primer M13F. The
contiguous sequence generated was deposited into GenBank
(Accession No. KX090269). Six of the seven clones contained an
Corresponding author: N K. Osterbauer. Email: nosterbauer@oda.state.or.us.

FIGURE 2
doi:10.1094 / PHP-BR-15-0055 Foliar symptoms caused by Lilac ring mottle virus on lilac ‘Krasavitsa
© 2016 The American Phytopathological Society Moskvy.’
PLANT HEALTH PROGRESS  Vol. 17, No. 3, 2016  Page 158

insert that showed >99% homology with the published sequence 1992). Fruit trees infected by Ilarvirus and other viruses often
for the RNA3 of LiRMoV (GenBank Accession No. U17391). exhibit an acute phase, which occurs when the plant is initially
Tomato mosaic virus (ToMV), Arabis mosaic virus (ArMV), infected and symptoms are obvious, and a chronic phase, which
and Lilac leaf chlorosis virus (LLVC) have also been described as occurs in subsequent growing seasons when systemic infection
infecting lilac (Cooper 1993). Additional PCR testing of the lilac has been established and plants may appear asymptomatic
samples’ TNA for these three viruses was completed. Amplicons (Nemeth 1986). It is possible LiRMoV, ToMV, and LLCV have
were produced from the lilac samples of the appropriate sizes for been present in these cultivars for many years as cryptic
LLCV (271 bp) (James et al. 2010) and for ToMV (318 bp, using infections. The infected plants at the Marion County nursery were
the primers 1180, 5′-CGAGAGGGGCAACAAACAT-3′, and destroyed.
1181, 5′-ACCTGTCTCCATCTCTTTGG-3′ [S. Scott
unpublished]); ArMV was not detected. Identity of the amplicons LITERATURE CITED
produced was confirmed by sequencing.
Samples received by the OSU Plant Clinic were examined Castello, J. D., Hibben, C. R., and Jacobi, V. 1992. Isolation of tomato mosaic
using transmission electron microscopy (TEM) at the OSU virus from lilac. Plant Dis. 76:696-699.
Electron Microscopy Facility, which used two methods to Cooper, J. I. 1993. Virus Diseases of Trees and Shrubs, 2nd ed. Chapman and
examine symptomatic leaves. In the first, the leaf tissue was cut Hall, London.
Favorite, J. 2006. Lilac: Syringa vulgaris. USDA Natural Resource Conserva-
into 1-mm strips and put in fixative (2.5% glutaraldehyde plus 1%
tion Plant Guide, Washington, DC. http://plants.usda.gov/core/
paraformaldehyde in sodium cacodylate buffer, 0.1M at 7.4 pH) profile?symbol=SYVU.
overnight. The following day the tissue was crushed to express Hughes, D. W., and Galau, G. 1988. Preparation of RNA from cotton leaves
sap, which was collected on a copper grid, stained with 2% and pollen. Plant Mol. Biol. Rep. 6:253-257.
aqueous ammonium phosphotungstic acid, and examined. The James, D., Varga, A., Leippi, L., Godkin, S., and Masters, C. 2010. Sequence
second method was a simple expression of sap from unfixed analysis of RNA2 and RNA3 of lilac leaf chlorosis virus: A putative new
member of the genus Ilarvirus. Arch. Virol. 155:993-998.
tissue, which was then stained with 2% uranyl acetate in 50%
Nemeth, M. 1986. Virus, Mycoplasma, and Rickettsia Diseases of Fruit Trees.
ethanol. Both preparations were examined using a FEI Titan 80- Martinus Nijhoff, Dordrecht, Netherlands.
200 TEM/STEM electron microscope. No virus particles were Scott, S. W., and Ge, X. 1995. The complete nucleotide sequence of the RNA
observed in either preparation. 3 of lilac ring mottle ilarvirus. J. Gen. Virol. 76:1801-1806.
This is the first report of LiRMoV from lilac cultivars President Scott, S. W., and Zimmerman, M. T. 2008. Partial nucleotide sequences of the
Grevy and Krasavitsa Moskvy in the United States. These RNA 1 and RNA2 of lilac ring mottle virus confirm that this virus should
be considered a member of subgroup 2 of the genus Ilarvirus. Arch. Virol.
cultivars were introduced into North America before 1950 and are 153:2169-2172.
grown in landscapes in 31 states and four Canadian provinces Van Der Meer, F. A., Huttinga, H., and Maat, D. Z. 1976. Lilac ring mottle
(Favorite 2006). LLCV was recently detected and described from virus: Isolation from lilac, some properties, and relation to lilac ringspot
Canada (James et al. 2010) and ToMV has previously been disease. Neth. J. Plant Pathol. 82:67-80.
reported infecting a lilac hybrid in New York (Castello et al.
PLANT HEALTH PROGRESS  Vol. 17, No. 3, 2016  Page 159

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Plant Health Brief

First Report of Lilac ring mottle virus Infecting

Accepted for publication 26 May 2016. Published 14 July 2016.

In May 2015, the Oregon Department of Agriculture investi-

Corresponding author: N K. Osterbauer. Email: nosterbauer@oda.state.or.us.

PLANT HEALTH PROGRESS  Vol. 17, No. 3, 2016  Page 158

PLANT HEALTH PROGRESS  Vol. 17, No. 3, 2016  Page 159

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