Turnip crinkle virus
Glasshouse Crops Research Institute, Littlehampton, Sussex, England
Olwen M. Stone
Glasshouse Crops Research Institute, Littlehampton, Sussex, England
Broadbent & Blencowe (1955), and
Broadbent & Heathcote (1958).
An RNA-containing virus with isometric particles about 28 nm in diameter. It occurs
in Brassica crops and is transmitted by flea-beetles and by inoculation of sap.
It is not widespread or common, although it has a fairly wide experimental host range.
Causes leaf crinkling, mottle and slight stunting in turnip,
or sometimes more severe distortion and rosetting.
Other species of Brassica
develop mild mottle
Recorded in Scotland, England and Yugoslavia.
Host Range and Symptomatology
It has a fairly wide host range, infecting
species in about 20 dicotyledonous families. Readily transmissible by inoculation of sap.
- Chenopodium amaranticolor. Numerous local chlorotic dots after 6 days, becoming
necrotic later; no systemic infection
- Chenopodium murale. Numerous local chlorotic dots after 10 days, expanding
to large water-soaked rings;
systemic chlorotic flecks after 3 weeks
- Chenopodium quinoa. Local chlorotic lesions after 10-14 days; no systemic
- Datura stramonium. Local chlorotic spots after 10 days; no systemic infection.
- Tetragonia expansa. Numerous local chlorotic dots after 10 days; symptomless
- Brassica pekinensis (Pe-tsai, Chinese cabbage). Local chlorotic diffuse
spots after 7 days; systemic light and dark green mottle and leaf crinkle
- Brassica pekinensis or B. juncea are good hosts for maintaining cultures
and as sources for purification.
- C. amaranticolor is a good local lesion host. C. quinoa is less
satisfactory; lesions are fewer, and take longer to appear.
None recorded; the virus reported by
Verma & Varma (1961)
proved to be turnip crinkle virus; the reactions in solanaceous plants suggest some
other virus, and no serological or cross-protection tests were done using the type
strain of turnip crinkle virus.
Transmission by Vectors
Transmitted by larvae and adult flea-beetles of the
(9 species) and Psylliodes
(2 species). The virus was
acquired within a few minutes and the beetles rarely remained infective for more than
There are unconfirmed reports of transmission by larvae of the butterfly Pieris
brassicae, leaf- mining larvae of the fly Phytomyza rufipes, hoppers of the
locust Locusta migratoria and four species of aphid
transmission using Myzus persicae was not confirmed
(Hollings & Stone, unpublished).
Transmission through Seed
Transmission by Dodder
No transmission by Cuscuta campestris
& Stone, unpublished).
The virus is a good immunogen; rabbits immunized by one intravenous
plus two intramuscular injections gave antisera with specific titres in precipitin
tube tests of 1/2048. The precipitates are granular (somatic). In gel-diffusion tests
a single reaction line is formed; good reactions are obtained with crude sap. In
immunoelectrophoresis, the single antigen component moves to the anode
(Hollings & Thorne, 1969
No serological relationships were found to any of twenty-seven
other isometric viruses
(Hollings & Stone, 1969
McLeod & Markham (1963)
suggested might be related
Stability in Sap
In turnip or Brassica pekinensis
sap the thermal
inactivation point is 90-95°C, dilution end-point more than 10-6
the virus survived at least 6 weeks at room temperature (c.
Lyophilized sap stored under vacuum at room temperature retained infectivity for at
least 9 years.
The following methods are satisfactory. Preparations from
may yield 250 mg virus per kg leaf tissue.
1. Hollings & Stone (1969).
Grind leaves at room temperature with 0.05 M phosphate
buffer (pH 7.6) containing 0.1% thioglycollic acid (wt/vol=1/1.25). Squeeze out sap.
Add n-butanol to 8.5% of the total volume and store at 2°C for 12-48 hr.
Concentrate the virus by differential centrifugation. Further purification can be done
by density gradient centrifugation.
2. Leberman (1966).
Harvest plants 4-6 weeks after infection. Mince leaves and strain
through muslin. Clarify by centrifuging at 12,000 g for 30 min. Add 1.34
g of a 20% solution of sodium dextran sulphate, 29 g of a 30% solution of polyethylene
glycol 6000 and 5 g of 5 M NaCl to each 100 ml of clarified sap. Pour into separating
funnel and leave to stand at 4°C overnight. Collect lower phase and interface, and
centrifuge at low speed; discard supernatant fluid. Add 3 M KCl to the pellet, stirring
until a thin creamy paste is obtained. Centrifuge at low speed; re-extract pellet with
0.5 M KCl if desired, and again centrifuge at low speed. Supernatant fluids are then
centrifuged at high speed (30,000 rev/min) for 2 1/2 hr and the pellet resuspended in
0.l M disodium ethylene diamine tetraacetate, pH 7. The virus can be further purified
by more cycles of high and low speed centrifugation, and resuspended in any of several
Properties of Particles
Sedimentation coefficient (s20, w
infinite dilution 129 S.
Molecular weight (daltons): 8.0-9.0 x 106.
Electrophoretic mobility: In immunoelectrophoresis tests using 0.9% Ionagar in 0.03
M phosphate buffer, pH 7.6, the virus migrates as a single antigenic component towards
the anode at a rate of 7.44 x 10-6 cm2 sec-1
Particles are isometric, about 28 nm in diameter in
phosphotungstate, 33 nm in uranyl acetate. The intact virus is not penetrated by
The particles contain 180 protein subunits of M. Wt 38,000 clustered
in dimers in the 2-fold positions of a T
=3 icosahedral lattice
(Finch, Klug & Leberman, 1970
In addition it is suggested that there are 12 subunits of M. Wt
28,000 probably situated on the 5-fold axes and, possibly, a small amount of protein
of M. Wt 80,000 which may represent the RNA polymerase
particles derived from the virus may represent inner cores
(Haselkorn et al., 1961
Finch et al., 1970
but they can be produced by re-aggregation of
the protein from degraded virus
(Leberman & Finch, 1970
About 17% of particle weight, probably single-stranded. Molar
percentages of nucleotides:
G28; A26; C24; U22
(Symons et al., 1963
Protein: About 83% of particle weight. The particles contain protein
components of M. Wt 38,000 and 28,000, and possibly a third of M. Wt 80,000
Amino acid compositions of the three proteins were determined by
Relations with Cells and Tissues
All tissues are infected; no inclusion bodies
reported, but plastid changes occur in some hosts
Other beetle-transmitted isometric viruses infecting Cruciferae are
turnip yellow mosaic
These three viruses cannot
be reliably distinguished from turnip crinkle virus by the symptoms produced in
species but they differ from turnip crinkle virus in that they do not
infect species of Chenopodium
or plants in families not close to the Cruciferae.
They are all serologically distinct from turnip crinkle virus. In addition, preparations
of turnip yellow mosaic and radish mosaic viruses separate into more than one component
on analytical or density gradient centrifugation.
- Broadbent, Investigation of viruses diseases of Brassica crops, Agric. Res. Council Rep. No. 14, Cambridge Univ. Press, 1957.
- Broadbent & Blencowe, Rep. Rothamsted exp. Stn. 1954: 87, 1955.
- Broadbent & Heathcote, Ann. appl. Biol. 46: 585, 1958.
- Butler, J. molec. Biol. 52: 589, 1970.
- Finch, Klug & Leberman, J. molec. Biol. 50: 215, 1970.
- Haselkorn, Hills, Markham & Rees, Abstr. Intern. Congr. Biophys. Stockholm 1961: 293, 1961.
- Hollings & Stone, Zentbl. Bakt. ParasitKde., Abt. II 123: 237, 1969.
- Hollings & Thorne, Rep. Glasshouse Crops Res. Inst. 1968: 109, 1969.
- Leberman, Virology 30: 341, 1966.
- Leberman & Finch, J. molec. Biol. 50: 209, 1970.
- McLeod & Markham, Virology 19: 190, 1963.
- Martini, Proc. 3rd Conf. Potato Virus Dis. Lisse-Wageningen 1957: 106, 1958.
- Stefanac, Acta biol. Iugoslav. Ser. B 6: 27, 1969.
- Symons, Rees, Short & Markham, J. molec. Biol. 6: 1, 1963.
- Tremaine, Virology 42: 611, 1970.
- Verma & Varma, Indian J. Microbiol. 1: 37, 1961.
Systemically infected plant of Brassica pekinensis.
Systemically infected leaf of B. pekinensis showing crinkling and
Virus particles from a purified preparation, stained with potassium
phosphotungstate. Bar represents 100 nm.
Systemically infected plant of B. rapa.
Local chlorotic lesions in Chenopodium amaranticolor.
Local chlorotic lesions in C. murale.
Immunoelectrophoresis; migration of virus towards anode (0.9% agar in
0.03 M phosphate buffer, pH 7.6, 4 hr at set voltage, 200 V).