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See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/274963501

First Report of Garlic virus A in Garlic From India

Article in Plant Disease · April 2015


DOI: 10.1094/PDIS-11-14-1210-PDN

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4 authors:

Suresh Gawande Vishal Gurav


Directorate Of Onion and Garlic Research Directorate Of Onion and Garlic Research
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Ashish Anil Ingle Jai Gopal


Directorate Of Onion and Garlic Research Indian Council of Agricultural Research
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Accepted for publication
http://dx.doi.org/10.1094/PDIS-11-14-1210-PDN

First report of Garlic virus A in garlic from


India
Dr. Suresh Janardhan Gawande, PhD
Directorate of Onion and Garlic Research, ICAR, Rajgurunagar, Pune, 410505, Pune,
Maharashtra,
India, 410505, +912135222026, +912135224056;[email protected]

Mr. V.S Gurav


Directorate of Onion and Garlic Research, Crop Protection, DOGR, Rajgurunagar, Pune
(MS), Pune, Maharashtra, India, 410505; [email protected]

Mr. Ashish A Ingle


Directorate of Onion and Garlic Research, Plant Pathology, Rajguru Nagar, Nasik
Road,Pune, Pune, Maharashtra, India, 410505;
Directorate of Onion and Garlic Research, Crop Protection, DOGR, Rajgurunagar, Pune
(MS), Pune, Maharashtra, India, 410505; [email protected]

Dr. J. Gopal
Directorate of Onion and Garlic Research, ICAR, Pune, Maharashtra, India;
Directorate of Onion and Garlic Research, Crop Protection, DOGR, Rajgurunagar, Pune
(MS), Pune, Maharashtra, India, 410505; [email protected]

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Garlic (Allium sativum L.) is an important spice crop in India and globally. Degeneration
due to latent viruses is an important constraint to garlic production. Mite-borne viruses,
members of the genus Allexivirus of the family Alphaflexiviridae are among latent viruses
known to commonly occur on Allium plants in many parts of the world. Garlic virus A
(GarV-A) is a prominent member of genus Allexivirus and is reported from many
countries (Cafrune et al. 2006; Dovas et al. 2001; Mohammed et al. 2013; Wylie et al.
2012). In August 2014, mild mosaic like symptoms were observed on the leaves of one
month old garlic cultivars Bhima Omkar, Bhima Purple, G-41 and G-282 at the
experimental plot of ICAR-Directorate of Onion and Garlic Research, Rajgurunagar, Pune,
India. Additionally, the presence of eriophyid mites on symptomatic plants lead to the
suspicion of possible Allexivirus infection in these cultivars. Total RNA from symptomatic
leaf samples (five samples per accession) was extracted using the RNeasy Plant Mini kit
(Qiagen GmbH, Hilden, Germany) and assayed by reverse transcription (RT)-PCR and
allexivirus specific primer pair ALLEX1 (5′-CYGCTAAGCTATATGCTGAARGG-3′) and
ALLEX2 (5′-TGTTRCAARGTAAGTTTAGYAATATCAACA-3′) (Dovas et al. 2001). All the
samples tested were positive for Allexivirus by RT-PCR. These samples were then tested
for individual allexiviruses such as GarV-A, Garlic virus B (GarV-B), Garlic virus C (GarV-
C), Garlic virus D (GarV-D) and Garlic virus X (GarV-X). Only GarV-A was detected using
the primer pair 5′-TGGAGACCCTTTCCAAGGCA-3′ and 5′-CTCATCCGCGAATGGTGGTT-3′
(Wylie et al. 2012) designed to amplify a 417 nt region within the triple gene block-1
(TGB-1) gene of GarV-A. A second set of primers DGAP1F (5′-
GACGGCTGTGCAAAAGCTTT-3′) and DGAP1R (5′-TGCATGCGTTTAATGAACAC-3′) was
used to generate a 936bp amplicon from the RNA-dependent RNA polymerase (RdRp)
gene of GarV-A. The amplified products from cultivar Bhima Omkar were purified using
the QIAquick PCR Purification Kit (Qiagen Inc., Hilden, Germany) and cloned in pDrive
(Qiagen Inc., Hilden, Germany). Two clones each of the TGB-1 and partial RdRp genes
were sequenced and a consensus sequence was determined and submitted to the
National Center for Biotechnology Information (NCBI) GenBank as accession numbers
KM588079 and KP835205, respectively. BLAST analysis revealed 80% nucleotide and
81% amino acid sequence identity of the TGB-1 sequence with Australian GarV-A
isolates (KF632716 and AGC09136), and 88% nucleotide and 98% amino acid sequence
identity of RdRp sequence with Australian GarV-A isolates (JX997952 and AGC09141).
To the best of our knowledge, this is the first report of GarV-A from India. Since, GarV-A
significantly reduces the garlic bulb yield (Cafrune et al. 2006), additional surveys and
evaluations are needed to study the magnitude of its impact on garlic production in
India. Reference: 1. E. E. Cafrune et al. Plant Dis. 90:898, 2006. 2. C.I. Dovas et al. J.
Phytopathol. 149:731, 2001. 3. H.S. Mohammed et al. J. Phytopathol. 161:642, 2013. 4.
S.J. Wylie et al. Arch. Virol. 157:271, 2012.

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