First report ofGroundnut bud necrosis virus
infecting onion (Allium cepa)
A. Sujitha &B. V. Bhaskara Reddy &Y. Sivaprasad &
R. Usha &D. V. R. Sai Gopal
Received: 30 December 2011 / Accepted: 11 October 2012 / Published online: 26 October 2012
#
Australasian Plant Pathology Society Inc. 2012
AbstractThe natural occurrence ofGroundnut bud necro-
sis virus(GBNV) on onion (Allium cepaL.) was detected by
enzyme linked immunosorbent assay using an antiserum
raised against GBNV and reverse transcription polymerase
chain reaction using coat protein gene specific primers.
Sequence analysis showed 93–100 % and 95–100 % iden-
tity at nucleotide and amino acid levels respectively with
other reported GBNV isolates.KeywordsGroundnut bud necrosis virus.
DAC-ELISA.
Onion.
RT-PCR
Onion (Allium cepaL.) is one of the most important vege-
table crops, which belong to the familyLiliaceae. Globally,
India occupies second position after China in production of
onion. In India it is grown in an area of 0.80 million ha with
a production of 13.37 million tonnes (www.faostat.com/
onion2010). It has several medicinal uses, including treat-
ment for diabetes, jaundice, fever, spleen enlargement etc.,
(Ross2001). More than 20 viruses of the generaTospovirus,
Potyvirus,CarlavirusandAllexivirusinfecting onion are
reported worldwide and cause severe yield losses (Maliogka
et al.2006).Tospovirusesare among the most damaging and
economically important group of plant viruses causing sig-
nificant crop losses in wide range of ornamental and food
crops in many regions of the World (Mumford et al.1996).
Groundnut bud necrosis virus(GBNV) (is also known as
Peanut bud necrosis virus) a member of the genusTospovi-
rus, familyBunyaviridaeconsists of enveloped, quasi spher-
ical particles, approximately 80–120 nm in diameter.
Among six serogroups ofTospovirus, only serogruop VI
(Iris yellow spot virus) has been reported to infect onion
crop (Ravi et al.2006). GBNV has a wide host range
infecting oil seed crops, vegetable crops, ornamental crops
and weeds (Reddy1991). The virus is transmitted by thrips
vector. Only ten species of thrips were reported as vectors of
A. Sujitha :B. V. Bhaskara Reddy (*) :Y. Sivaprasad
Department of Plant Pathology,
Regional Agricultural Research Station,
Tirupati 517502, Andhra Pradesh, India
e-mail: drbhaskarareddy@gmail.com
A. Sujitha
e-mail: sujithaasadhi@gmail.com
Y. Sivaprasad
e-mail: yeturusivaprasad@gmail.com
R. Usha
Department of Biotechnology, Sri Padmavati Mahila
Visvavidyalayam,
Tirupati 517502, Andhra Pradesh, India
e-mail: waves.sea@gmail.com
D. V. R. Sai Gopal
Department of Virology, Sri Venkateswara University,
Tirupati 517502, Andhra Pradesh, India
e-mail: profdvrsaigopalift@gmail.com Australasian Plant Dis. Notes (2012) 7:183–187
DOI 10.1007/s13314-012-0080-8
tospoviruses recorded Worldwide (Ullman et al.1997). An
earlier study from India detected the presence ofTospovirus
inS.dorsalis(Meena et al.2005).During November 2011 in commercial onion fields in the
Kadapa district of Andhra Pradesh, India, Straw coloured,
mosaic and necrotic lesions were observed on the young
Fig. 1Symptoms associated
with natural occurrence of
Groundnut bud necrosis virus
on onion: mosaic, chlorotic
spots, necrosis of young leaves
Fig. 2Agarose gel
electrophoresis of RT-PCR
products. Lane M, 1Kb DNA
ladder, Lane 1, uninfected.
Lane 2, 3, 4, infected onion
samples
184A. Sujitha et al.
Ta b l e 1Sequence identity of present isolate (JQ269832) at nucleotide (above the diagonal) and amino acid (below the diagonal) levels
respectively with other reported GBNV isolates
Isolates JQ269832 HQ199845 HQ199844 HQ324113 HQ324115 EF179100 FJ447359
JQ269832 100 99.7 99.8 99.8 97.4 99.3 98
HQ199845 99.2 100 99.8 99.8 97.4 99.3 98
HQ199844 99.6 99.6 100 100 97.5 99.5 98.1
HQ324113 99.6 99.6 100 100 97.5 99.5 98.1
HQ324115 99.2 99.2 99.6 99.6 100 97.3 97.2
EF179100 99.2 99.2 99.6 99.6 100 100 98.4
FJ447359 99.2 99.2 99.6 99.6 100 100 100
EF532937 97.8 97.8 98.1 98.1 98.5 98.5 98.5
HM770020 99.6 99.6 100 100 99.6 99.6 99.6
AY184354 97.8 97.8 98.1 98.1 98.5 98.5 98.5
AY512650 97.8 97.8 98.1 98.1 98.5 98.5 98.5
AY463968 95.6 95.6 96 96 96.3 96.3 96.3
DQ058078 95.6 95.6 96 96 96.3 96.3 96.3
AY882003 99.2 99.2 99.6 99.6 100 100 100
AF467289 98.1 98.1 98.5 98.5 98.9 98.9 98.9
AY529713 98.1 98.1 98.5 98.5 98.9 98.9 98.9
AF515821 98.9 98.9 99.2 99.2 99.6 99.6 99.6
AY512648 98.9 98.9 99.2 99.2 99.6 99.6 99.6
AY512651 98.5 98.5 98.9 98.9 99.2 99.2 99.2
AY512647 97.8 97.8 98.1 98.1 98.5 98.5 98.5
AY426317 95.2 95.2 95.6 95.6 96 96 96
Isolates EF532937 HM770020 AY184354 AY512650 AY463968 DQ058078 AY882003
JQ269832 97.4 99.6 97.4 97.5 94.1 94.3 97.1
HQ199845 97.4 99.6 97.4 97.5 94.1 94.3 97.1
HQ199844 97.5 99.7 97.5 97.7 94.2 94.4 97.2
HQ324113 97.5 99.7 97.5 97.7 94.2 94.4 97.2
HQ324115 96.8 97.3 96.8 96.9 94.5 94.8 97.1
EF179100 97.5 99.7 97.5 98 94.2 94.4 97.7
FJ447359 97.7 98.1 97.7 98 94.3 94.5 97.2
EF532937 100 97.3 98.1 98.1 94.4 94.7 96.6
HM770020 98.1 100 97.3 97.8 93.9 94.2 97.4
AY184354 98.1 98.1 100 98 94.3 94.5 96.6
AY512650 98.5 98.1 98.1 100 94.5 94.5 97.1
AY463968 96 96 95.6 96 100 99.5 93.9
DQ058078 96 96 95.6 96 100 100 94.2
AY882003 98.5 99.6 98.5 98.5 96.3 96.3 100
AF467289 97.4 98.5 97.4 97.4 95.2 95.2 98.9
AY529713 98.9 98.5 98.5 98.9 96.3 96.3 98.9
AF515821 98.1 99.2 98.1 98.5 96 96 99.6
AY512648 98.1 99.2 98.1 98.1 96 96 99.6
AY512651 99.2 98.9 98.9 99.2 96.7 96.7 99.2
AY512647 98.5 98.1 98.1 98.5 96 96 98.5
AY426317 95.6 95.6 95.2 95.6 99.2 99.2 96
Isolates AF467289 AY529713 AF515821 AY512648 AY512651 AY512647 AY426317
JQ269832 97.2 97.1 98.3 98.1 97.3 97.3 93
HQ199845 97.2 97.1 98.3 98.1 97.3 97.3 93
HQ199844 97.3 97.2 98.4 98.3 97.4 97.4 93
HQ324113 97.3 97.2 98.4 98.3 97.4 97.4 93
HQ324115 96.6 96.2 97.2 97.1 96.6 96.7 94
EF179100 97.3 97.4 98.4 98.7 97.9 97.9 93
FJ447359 97.9 97.1 98 98.1 97.7 97.8 94
EF532937 96.8 97.4 97.7 97.8 97.8 97.9 94
HM770020 97.1 97.2 98.1 98.5 97.7 97.7 93
First report ofGroundnut bud necrosis virusinfecting onion185
leaves. The necrosis starts with apical portion of young
leaves and flower stalks and finally flower abortion and
plant death (Fig.1). Based on above symptomatology
GBNV infection was suspected. The presence of virus in
symptomatic leaves was confirmed by direct antigen coating(DAC)-ELISA (Clark and Joseph1984) using GBNV poly-
clonal antibodies. RT-PCR tests of leaf tissue from onion
plants using primers specific for the nucleocapsid gene of
GBNV (Satyanarayana et al.1996) resulted in an amplicon
of the expected size (~800 bp) (Fig.2). The amplicon
Table 1(continued)
Isolates AF467289 AY529713 AF515821 AY512648 AY512651 AY512647 AY426317
AY184354 96.8 97.5 97.7 97.5 97.7 97.8 94
AY512650 96.9 98.1 97.8 98 98.3 98.4 94
AY463968 93.7 94.5 94.3 94.2 94.4 94.1 98
DQ058078 93.9 94.5 94.8 94.4 94.7 94.3 98
AY882003 96.3 96.7 97.2 97.8 97.1 96.9 93
AF467289 100 96.5 97.2 97.1 96.6 96.7 93
AY529713 97.8 100 97.3 97.7 97.4 97.5 94
AF515821 98.5 98.5 100 98.4 97.4 97.8 94
AY512648 98.5 98.5 99.2 100 97.8 97.9 94
AY512651 98.1 99.6 98.9 98.9 100 98.1 94
AY512647 97.4 98.9 98.1 98.1 99.2 100 94
AY426317 94.9 96 95.6 95.6 96.3 95.6 100
Fig. 3Phylogenetic tree using MEGA 4.0 obtained from Clustal W alignments of the coat protein gene of Onion-Kadapa (JQ269832) with the
other GBNV isolates
186A. Sujitha et al.
(PBNV-Kadapa) was cloned into pTZ57R/T vector (Fermen-
tas, USA), sequenced and deposited in GenBank DNA data-
base (Accession No. JQ269832). Sequence analysis (BioEdit
v. 7.05) and comparison with other GBNV isolates (GenBank
Accession No. HQ199845, HQ199844, HQ324113,
HQ324115, EF179100, FJ447359, EF532937, HM770020,
AY184354, AY512650, AY463968, DQ058078, AY882003,
AF467289, AY529713, AF515821, AY512648, AY512651,
AY512647 and AY426317) showed 93–100 % and 95–100 %
identity at nucleotide and amino acid levels respectively
(Table1). Phylogenetic tree at nucleotide level was constructed
using MEGA version 4.0 and it further showed that the present
isolate (Onion-Kadapa-JQ269832) is closely related to Taro-
NLR (HQ199845), Calotropis-NLR (HQ199844), Cotton-
TPT (HQ324113) and Groundnut-Coimbatoor (HM770020)
(Fig.3).
The infected crop has now been removed to eradicate the
infection. An earlier study confirmed the prevalence of
IYSV (Tospovirus: Serogroup VI) on onion in India (Ravi
et al.2006). This is the first report of the natural occurrence
of GBNV (Serogroup IV ofTospovirus) infecting onion.
AcknowledgmentsWe a r e v e r y g r a t e f u l t o D r. P. L a v a K u m a r,
International Institute of Tropical Agriculture Ibadan for his gift of
antisera.
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First report ofGroundnut bud necrosis virusinfecting onion187
