Apple stem grooving virus
Faculty of Agriculture, Iwate University, Morioka 020-8550, Japan.
Lister et al. (1965) from apple,
Wallace & Drake (1962) from citrus, and
Inouye et al. (1979) from lily.
- Selected Synonyms
- Apple latent virus Type 2 (C-431 isolate)(
Lister et al., 1965)
- Dark green epinasty virus (in Chenopodium quinoa) (
Waterworth & Gilmer, 1969)
- Citrus tatter leaf virus (
Wallace & Drake, 1962)
- Citrange stunt virus (
Wallace & Drake, 1968;
A virus with flexuous filamentous particles 620-650 long and 12nm in diameter containing
linear positive-sense, ssRNA. Probably occurs world-wide in Rosaceae fruit trees and
citrus in which it is usually symptomless. Its natural mode of spread is unknown. It is
symptomlessly in most Rosaceae fruit trees and citrus plants. Transmissible by
mechanical inoculation to herbaceous plants.
Causes stem grooving, brown line and graft union abnormalities in Virginia Crab
topworking disease of apple trees grown on Mitsuba kaido (Malus sieboldii
) in Japan (
and bud union abnormalities of citrus trees on trifoliate orange
(Calavan et al., 1963
Miyakawa & Matsui, 1976
Probably distributed wherever apples are cultivated. Natural spread in citrus plants is reported in
China and Japan
(Ke & Wu, 1991
Miyakawa & Matsui, 1976
Su & Cheon, 1984
Zhang et al., 1988
Host Range and Symptomatology
Infects the Rosaceae fruit trees, apple, European pear, Japanese pear, and
Japanese apricot and citrus plants. Infects experimentally more than 40 species in 17
families (Aizoaceae, Amaranthaceae, Apocynaceae, Caryophyllaceae, Chenopodiaceae,
Compositae, Convolvulaceae, Cucurbitaceae, Graminae, Labiatae, Leguminosae, Liliaceae,
Pedaliaceae, Rosaceae, Rutaceae, Scrophulariaceae, Solanaceae)
(Inouye et al., 1979;
Lister et al., 1965;
Semancik & Weathers, 1965;
Waterworth & Gilmer, 1969),
many symptomlessly. Transmissible from apple to C. quinoa by mechanical inoculation
extracts from buds, young leaves or petals ground in 0.05M phosphate buffer (pH7-8)
containing 2% (v/v) nicotine base or 2% (w/v) polyvinyl pyrrolidone
- Diagnostic species
Malus pumila cv. Virginia Crab (apple). Stem grooving in the wood.
Necrosis and pitting at the union of Viriginia Crab and rootstock (Fig.1).
M. sieboldii "MO65". Necrosis and pitting at the scion-rootstock
Citrus excelsa. Tatter leaf (Fig.4).
Citrange (Poncirus trifoliata x C. sinensis) cvs. Rusk and Troyer. Leaf
malformation and chlorotic mottling (Fig.5).
P. trifoliata and citrange. Bud-union crease between infected scion and
Chenopodium quinoa. Systemic leaf epinasty, distortion, and mottle
and stunting. Some isolates induce necrotic lesions 0.5-2 mm in diameter in inoculated
Nicotiana glutinosa. Systemic conspicuous yellow to mild mosaic symptoms
depending on the isolate.
Phaseolus vulgaris cv. Pinto. Purple-brown spots or rings 0.5-3 mm in diameter
or chlorotic spots on the inoculated leaves. Some isolates induced veinal necrosis
in inoculated leaves. Symptoms are generally were more severe in winter.
- Propagation species
C. quinoa is the most useful plant for propagation. Zucchini squash is also
reported as a propagation host for some isolates
(Uyemoto & Gilmer, 1971).
To transmit the virus from fruit trees, N. occidentalis, a symptomless host is
N. glutinosa can be used to eliminate contaminating viruses from apple such as
Apple chlorotic leaf spot virus and
Tobacco mosaic virus.
- Assay species
C. quinoa and P. vulgaris cv. Pinto are useful local
Many isolates are reported from apple, Japanese pear, European pear, Japanese apricot, lily and
citrus plants, but most are not well characterized biologically and serologically. Some isolates
have been differentiated only on symptomatology
This virus and Citrus tatter leaf virus (CTLV) have been regarded as distinct
viruses. However, CTLV from citrus and lily is indistinguishable from ASGV from Rosaceae
fruit trees biologically, serologically, in genome organization and in nucleotide sequence
(Kawai et al., 1991;
Magome et al., 1997a;
Ohira et al., 1995;
Yoshikawa et al., 1993;
Isolate Li-23 from lily can infect citrus plants (N. Inouye, personal communication) and isolate V-3
from Japanese pear induced symptoms similar to those by CTLV from citrus on Rusk citrange
(Iwanami et al. 1991).
From these results, CTLV is regarded as an isolate of ASGV.
Virus isolates from apple, Japanese pear and European pear trees comprise at least two to four
variants that differ considerably from each other in nucleotide sequence
(Magome et al., 1997a;
Yoshikawa et al., 1996).
The composition of sequence variants within a tree differed among leaves from different branches,
showing that each sequence variant is distributed unevenly within an individual tree
(Magome et al., 1999).
Transmission by Vectors
No vectors are reported.
Transmission through Seed
Transmitted through seed to progeny seedlings of lily (1.8%) and C. quinoa
(2.5 - 60.0%)
(Inouye et al., 1979
Moderately antigenic, giving antisera from rabbits with titers of 1/128 to 1/4,096.
Polyclonal antisera have been prepared in rabbits against purified virus or coat protein expressed
in Escherichia coli
(de Sequeira & Lister, 1969a
Inouye et al., 1979
Nishio et al., 1989
Yanase et al., 1986
ELISA has been used to detect the virus in fruit trees
Namba et al., 1992
Yanase et al., 1986
Monoclonal antibodies were produced in mice against an isolate from citrus; three selected
monoclonal antibodies react with all isolates tested, including 9 isolates from citrus in
Japan, 4 isolates from citrus in the USA, 6 isolates from Chinese citrus, and isolates from lily,
apple and Japanese apricot
(Kawai et al., 1991
Serologically unrelated to all known virus species in the genus Capillovirus
Similarities exist between the virus and virus species in the genera Trichovirus
in the amino acid sequences of conserved polymerase motif, putative
movement protein and coat protein
Yoshikawa et al., 1992
Stability in Sap
In bentonite-clarified C. quinoa
sap, the thermal inactivation point (10 min) is
between 60 and 63°C, the half-life at 45°C is 105±8 min, the dilution end-point is
, and infectivity is retained for more than 2 days at 20°C and more
than 27 days at 4°C
In crude sap of infected C. quinoa
, an isolate from lily has a thermal inactivation point
between 65 and 70°C, a dilution end-point between 10-4
longevity in vitro
between 4 and 8 days at 20°C
(Inouye et al., 1979
Purified from infected C. quinoa
(de Sequeira & Lister ,1969b
by homogenizing 100g leaves in 0.01M or 0.05M phosphate buffer (pH7-8) containing 0.15%
sodium thioglycollate or 1% 2-mercaptoethanol. Clarify the extract by low speed centrifugation
and squeeze through cheesecloth. Add a bentonite suspension (30-40 mg/ml in 0.01M phosphate
buffer, pH7-8) slowly and clarify by centrifuging at low speed. Repeat this clarification step
until the supernatant fluid is clear. Precipitate the virus from the supernatant fluid by adding
PEG (mol. wt 6,000) to 4-8 %(w/v) and NaCl to 0.02M, incubate for 1 hr and centrifuge at low
speed. Resuspend the pellet in 0.01M phosphate buffer (pH7.0) and clarify by centrifuging at low
speed. Concentrate the virus by ultracentrifugation and purify further by sucrose density
gradient centrifugation. Yields are 50 - 250 µg/100g leaf tissue.
Properties of Particles
Sedimentation coefficient, S20,w
112 at 0.3-0.6 mg/ml
in 0.01M neutral phosphate buffer.
Isoelectric point; c. pH 4.3 at ionic strength of 0.1.
Electrophoretic mobility; -10.3 and -6.5 X 10-5 cm2. sec-1.
V-1 respectively at pH 7.0 and pH 6.0 (ionic strength = 0.1).
A260/A280 : 1.18 not corrected for light-scattering
Particles are very flexuous filaments, 619±14 nm long and c.12 nm in width, with obvious
cross-banding and helical symmetry
) and a pitch of c. 3.8 nm.
Particle CompositionNucleic acid
: Linear positive-sense ssRNA of Mr c.
2.30 x 106
or 6,496 nucleotides excluding a polyA-tail
(Yoshikawa & Takahashi, 1988
Yoshikawa et al., 1992
about 5% of particle weight. Nucleotide base ratios for isolate P-209 from apple are:
G 23.0%; A 30.6%; C 18.4%; U 28.0%
(Yoshikawa et al., 1992
Protein: Particles contain a single polypeptide species of Mr 27,000
(Nishio et al., 1989;
Yoshikawa & Takahashi, 1988).
The complete nucleotide sequences (6496 bases) of the single RNA genome of three
been determined. They are isolates P-209
(accession no. D14995) from apple, L
(D16681) and Li-23
(AB004063) from lilies
(Yoshikawa et al., 1992;
Ohira et al., 1995;
Terauchi et al., 1997).
Identities of the nucleotide sequences were 82.9% (P-209/L), 83.0% (P-209/Li-23) and 98.4%
(Terauchi et al., 1997).
The genomic RNA has two overlapping ORFs in the positive strand
ORF1 (bases 37-6341) encodes a 241- to 242 kDa polyprotein (2105 amino acids) containing
consensus motifs of methyltransferase, papain-like protease, nucleotide triphosphate-
helicase, RNA polymerase, and coat protein (CP) in the C-terminal region. ORF2 (bases
encodes a 36 kDa putative movement protein (320 aa). A region (designated V-region) (aa
position 1585-1868) of the ORF1-encoded protein between the polymerase and the CP, that
ORF2 in another frame does not have any functional motifs found in other known plant virus
This region shows high variability among isolates and sequence variants
(Magome et al., 1997a).
Although the CP is located in the C-terminal region of the ORF1-encoded polyprotein and
RNA directed the synthesis of a polypeptide of c. 200 kDa as a major product in
in vitro translation, and was immunoprecipitated by antiserum to virus particle
(Yoshikawa & Takahashi, 1992),
the following evidence suggests that the CP is expressed from a subgenomic RNA
(Magome et al., 1997b).
Analysis of dsRNA from infected tissues indicate that all isolates tested contain five
virus-specific dsRNAs (6.5, 5.5, 4.5, 2.0 and 1.0 kbp). The 6.5 kbp species represents the
double-stranded form of the full-length genome, whereas the 2.0 and the 1.0 kbp species
the double-stranded forms of subgenomic RNAs coding for the putative movement protein and
CP, respectively. The size of the E. coli -expressed protein corresponding to the
C-terminal region of the ORF1-encoded protein, which starts with the Met at aa position
agreed with that of the CP.
Relations with Cells and Tissues
In infected C. quinoa
leaves, the particles occur singly or as aggregates in the cytoplasm
of mesophyll and phloem parenchyma cells. No virus-specific inclusion bodies, such as
pinwheels, viroplasmas or vesicles were observed
(Ohki et al., 1989
Ecology and Control
Because the virus is thought to be transmitted in the field only by grafting, planting virus-free
plants is the best means of controlling decline problems in apple and citrus plants due to the virus.
- Calavan, Christiansen & Roistacher, Plant Disease Reporter 47: 971, 1963.
- de Sequeira & Lister, Phytopathology 59: 572, 1969a.
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Necrosis at the union of Virginia Crab and an infected rootstock (right ) compared with a
healthy union (left) (Courtesy I. Machida).
Necrosis at the unions of M. sieboldii "M065" and infected rootstocks
(Courtesy I. Machida).
Basal scion swelling "Dai-Make" , at a bud-union of infected Satsuma Mandarin on
P. trifoliata rootstock (Courtesy T. Miyakawa).
Tatter symptoms on young leaves of C. excelsa (Courtesy T. Miyakawa).
Leaf distortion and chlorotic mottling of citrange cv. Rusk (Courtesy T. Miyakawa).
A bud-union crease between infected scion (Satsuma Mandarin) and rootstock
(P. trifoliata) (right) and a healthy bud-union (left)
(Courtesy T. Miyakawa).
Epinasty, distortion and mottling of a systemically infected C. quinoa plant.
Particles of isolate P-209 stained with uranyl acetate. Bar represents 100nm.
Genome organization of Apple stem grooving virus . MET, methyltransferase; P-PRO,
papain-like protease; HEL, helicase ; POL, RNA polymerase; MP, putative movement protein; V,
variable region; CP, coat protein.