Tobacco rattle virus
B. D. Harrison
Scottish Horticultural Research Institute, Invergowrie, Dundee, Scotland
- (Most of the names listed below refer to isolates that differ to some extent
from the type culture.)
- Aster ringspot virus (Rev. appl. Mycol. 33: 675)
- Belladonna mosaic virus (Rev. appl. Mycol. 22: 451)
- Potato corky ringspot virus (Rev. appl. Mycol. 26: 76; 37:
- Potato stem mottle virus (stengelbont virus) (Rev. appl. Mycol.
- Ratel virus (Rev. appl. Mycol. 25: 83)
- Tabakmauche Virus
and Rev. appl. Mycol. 37:
- Tabak Streifen und Kräuselkrankheit Virus (Rev. appl. Mycol.
- (For the major variant, pepper ringspot virus, see under Strains)
An RNA-containing virus with straight tubular particles of two predominant
lengths, the longer c. 190 nm and the shorter 45-115 nm, depending on
the isolate. Some isolates are readily transmitted by inoculation of sap but
others are not. The virus has a wide host range and is transmitted by soil-inhabiting
nematodes (Trichodorus spp.).
Causes one type of spraing (corky ringspot, Pfropfenbildung)
stem-mottle, in potato, notched leaf in gladiolus, ringspot in aster and pepper,
yellow blotch in sugar beet, and unnamed diseases in lettuce, hyacinth, narcissus,
tulip and several other ornamental species.
Europe, USA, Brazil, Japan.
Host Range and Symptomatology
The host range is very wide. More than 400 species in more than 50
dicotyledonous and monocotyledonous families can be infected; in many the virus
does not become systemic
Uschdraweit & Valentin, 1956
- Chenopodium amaranticolor. Necrotic local lesions
to spread; not systemic.
- Cucumis sativus (cucumber). Chlorotic or necrotic local lesions; not
- Nicotiana clevelandii. Inoculated leaves remain symptomless, or develop
chlorotic or necrotic lesions. Systemically infected leaves develop few symptoms or
variable amounts of necrotic flecking and distortion.
- Phaseolus vulgaris (French bean). Pin-point, necrotic local lesions
develop in 1-3 days
- Nicotiana clevelandii is the best host for maintaining cultures and as a
source of virus for purification.
- Chenopodium amaranticolor and Phaseolus vulgaris are the most useful
plants for local lesion assay. Nicotiana tabacum (tobacco,
Fig.1) and N.
can be used with isolates that produce local lesions in these
species. N. tabacum and Cucumis sativus are useful bait plants for
testing transmission by vectors.
Many strains are described but few are reliably distinguished by symptoms in test
plants. Some of the best characterized are:
(Cadman & Harrison, 1959;
Harrison & Nixon, 1959).
obtained from potato in Scotland. Now used as the type strain. Short particles
c. 75 nm long.
(Harrison & Woods, 1966)
= pepper ringspot virus
(Kitajima & Costa, 1969).
Originally obtained from Bidens pilosa in Brazil. Infects
N. tabacum without producing necrotic local lesions. Short particles 52 nm long.
Originally obtained from Capsicum
frutescens in California. Short particles 45 and 90 or 110 nm long.
Oregon strains. Originally obtained from potato in Oregon. Separated and
Lister & Bracker (1969).
One variant produces yellow ringspots
in N. clevelandii, N. glutinosa, etc. Short particles 48 and 81, 90 or 100 nm
Isolates unable to produce nucleoprotein particles (= NM = unstable isolates) can
be obtained from any of the above strains by using inocula containing only 190 nm
particles. They are poorly transmissible by mechanical inoculation unless inocula
are made using phenol. They cause more necrosis in plants than do their parent
and are slow to become systemic
are found in naturally infected plants.
Transmission by Vectors
At least 11 Trichodorus
spp. (stubby root nematodes) are natural vectors in
Europe or the USA but there is some evidence of specificity between virus strain and
(van Hoof, 1968
Taylor & Cadman, 1969
seem the most important vectors in Europe. Adults and larvae can
transmit. The virus can be acquired by T. allius
in 1 hr, inoculated in 1 hr
and retained for 20 weeks by nematodes kept in soil without plants
(Ayala & Allen, 1968
Transmission through Seed
Occurs (1-6%) in some weeds, for example Capsella bursa-pastoris
(Lister & Murant, 1967
Transmission by Dodder
At least 6 Cuscuta
spp. can transmit. The virus infects the dodder
Some strains are poorly immunogenic but antisera with titres of 1/1000 can be
made against others. Precipitin tests in mixed liquids have been used most. One of
two bands of precipitate develop in double diffusion tests in 1% agar gel.
Antigenic differences are found between some isolates from W. Europe, USA
and Japan, but isolates from Brazil differ more
(Harrison & Woods, 1966
distant serological relationship to pea early-browning virus was reported by
Stability in Sap
In N. clevelandii
sap, the thermal inactivation point (10 min) of several
strains is 80-85°C, dilution end-point about 10-6
and infectivity is
retained at 20°C for more than 6 weeks. By contrast, RNA-producing isolates lose
infectivity when sap is heated for 10 min at 60°C, diluted to 10-2
kept at 20°C for 3 h
Systemically infected N. clevelandii
leaves yield 20-100 mg virus per
1. Store sap at -15°C. Thaw overnight at 20°C then purify by low- and
high-speed centrifugation. Virus in pellets from the first high-speed centrifugation
should be allowed to resuspend in 0.07 M phosphate buffer (pH 7.5) for 16 h at
2. Grind cooled leaves in 0.01 M citric acid + phosphate buffer (pH 7.4, containing
0.1% sodium thioglycollate). Clarify extract by blending with 0.5 vol. of a 1:1 mixture
of n-butanol + chloroform and freeze aqueous layer overnight. Thaw and purify
virus by differential centrifugation, resuspending virus in 0.01 M phosphate buffer
(Lister & Bracker, 1969).
Properties of Particles
All strains (except RNA-producing isolates) produce tubular
particles of two or three predominant lengths, one c.
190 nm (L) and the
other(s) 45-115 nm (S),
depending on the isolate
L particles are infective and induce synthesis
of L-particle RNA.
S particles are non-infective alone but carry the gene for the protein found in the
L and S are produced only when the inoculum contains both L and S or their RNAs
Frost, Harrison & Woods, 1967
Lister & Bracker, 1969
Sedimentation coefficient (s20, w): c. 300 S
(L), 155-243 S (S).
Molecular weight (daltons): c. 50 x 106 (L), c. 12-29
x 106 (S).
Electrophoretic mobility: -1.7 x 10-5 cm2 sec-1
volt-1 in 0.067 M phosphate buffer, pH 7.0 (L and S, PRN strain).
Absorbance at 260 nm (1 mg/ml, 1 cm light path): c. 3.0.
Ultraviolet radiation inactivates nucleoprotein particles irreversibly but some
of the damage to extracted RNA is photoreactivable.
Particles are helically constructed. The pitch of the helix is 2.5 nm and it has a
central canal of diameter c.
5 nm. Number of protein subunits per turn = n + 1/3
(Nixon & Harrison, 1959
: Single-stranded; molecular weight 2.3 x 106
(L particles) and
0.6-1.3 x 106
(S particles); about 5% of particle weight. Sedimentation
coefficients are 26 S (L) and c.
12-20 S (S) in pH 7.4 buffer
(0.01 M Tris + 0.01 M KCl + 10-4
). Molar percentages of
G 25; A 29; C 17; U 29 (California isolate)
(Semancik & Kajiyama, 1967).
Protein: Subunits have a molecular weight of 2.4 x 104 and
contain about 218 amino acid residues
(Offord & Harris, 1965).
Amino acid composition
of two California isolates is given by
Relations with Cells and Tissues
Most tissues of some species become infected. L particles of strain CAM become
radially arranged around mitochondria
whereas S particles are dispersed
in the cytoplasm
(Harrison & Roberts, 1968
Kitajima & Costa, 1969
strains behave differently
(de Zoeten, 1966
In N. clevelandii
largely composed of abnormal mitochondria can be induced by a RNA-producing isolate
and by strain PRN
(Harrison, Stefanac & Roberts, 1970
Tobacco rattle virus mainly occurs on light sandy or peaty soils, which are the
preferred habitat of its nematode vectors. It may be patchily distributed within a
field. It can be distinguished from
pea early-browning virus
by failure to infect
pea systemically, by having slightly shorter L particles and by serological tests.
Also, most isolates of pea early-browning virus produce large lesions in Phaseolus
- Ayala & Allen, J. Agric. Univ. P. Rico 52: 101, 1968.
- Behrens, Landwn Vers Stnen 52: 442, 1899.
- Cadman, Nature, Lond. 193: 49, 1962.
- Cadman & Harrison, Ann. appl. Biol. 47: 542, 1959.
- de Zoeten, Phytopathology 56: 744, 1966.
- Frost, Harrison & Woods, J. gen. Virol. 1: 57, 1967.
- Harrison & Nixon, J. gen. Microbiol. 21: 569, 1959.
- Harrison & Roberts, J. gen. Virol. 3: 121, 1968.
- Harrison & Woods, Virology 28: 610, 1966.
- Harrison, Stefanac & Roberts, J. gen. Virol. 6: 127, 1970.
- Kitajima & Costa, J. gen. Virol. 4: 177, 1969.
- Lister, Virology 28: 350, 1966.
- Lister & Bracker, Virology 37: 262, 1969.
- Lister & Murant, Ann. appl. Biol. 59: 49, 1967.
- Maat, Neth. J. Pl. Path. 69: 287, 1963.
- Nixon & Harrison, J. gen. Microbiol. 21: 582, 1959.
- Offord, J. molec. Biol. 17: 370, 1966.
- Offord & Harris, Fed. European Biochem. Soc. 2nd meeting, abstracts: 216, 1965.
- Quanjer, Tijdschr. PlZiekt. 49: 37, 1943.
- Sänger, J. Virol. 3: 304, 1969.
- Schmelzer, Phytopath. Z. 28: 1, 1956.
- Schmelzer, Phytopath. Z. 30: 281, 1957.
- Semancik, Phytopathology 56: 1190, 1966.
- Semancik & Kajiyama, Virology 33: 523, 1967.
- Taylor & Cadman, in Viruses, Vectors and Vegetation, New York, Interscience Publishers, p. 55, 1969.
- Uschdraweit & Valentin, NachrBl. dt. PflSchutzdienst, Braunschweig 8: 132, 1956.
- van Hoof, Nematologica 14: 20, 1968.
Photographs: courtesy of the Scottish Horticulltural Research Institute.
Nicotiana tabacum cv. White Burley leaf inoculated with strain PRN.
Chenopodium amaranticolor leaf inoculated with strain CAM.
N. glutinosa leaf inoculated with strain PRN.
Naturally infected potato (Solanum tuberosum cv. Pentland Dell)
tuber cut to show spraing symptoms.
Part of Phaseolus vulgaris leaf inoculated with strain PRN.
N. clevelandii plant systemically infected with a RNA-producing
isolate derived from strain CAM.
L particles of strain CAM arranged radially around a mitochondrion in a
N. clevelandii leaf cell. Bar represents 200 nm.
Particles of strain CAM in phosphotungstate. Bar represents 100 nm.