New Disease Reports (2015) 32, 31. http://dx.doi.org/10.5197/j.2044-0588.2015.032.031
First report of Pepper mild mottle virus infecting chilli
pepper in Pakistan
A. Ahmad
1
*, A. Tiberini
2
, M. Ashfaq
1
and L. Tomassoli
3
1
Plant Virology Laboratory, Department of Plant Pathology, PMAS-Arid Agriculture University. Rawalpindi, Pakistan;
2
Università degli Studi "Mediterranea" di Reggio Calabria, Dipartimento di Agraria, 89122 Feo di Vito, Reggio Calabria, Italy;
3
Consiglio per la ricerca in agricoltura e l'analisi dell'economia agrari - CREA, Centro di ricerca per la patologia vegetale,
Via Carlo Giuseppe Bertero 22, 00156 Rome, Italy
*E-mail: adnan_84@outlook.com
Received: 25 Nov 2015. Published: 12 Dec 2015. Keywords: Capsicum annuum, tobamovirus
Figure 1
Chilli pepper (Capsicum annuum) is an important cash crop in Pakistan
which is cultivated on an area of 62,500 ha with an annual yield of 145,100
tonnes (Farooq, 2014). Pepper mild mottle virus (PMMoV) is a single-
stranded RNA virus belonging to the genus Tobamovirus, family
Virgaviridae (King et al., 2011). The virus possesses a serious threat to
pepper cultivation worldwide due to its long-term survival and highly
efficient transmission through seeds, plant debris, and on contaminated
worker’s hands and tools.
During the summers of 2013 and 2014, surveys were done to identify the
major viruses infecting chilli crops in the Pothwar region of Pakistan. A
total of 25 chilli pepper leaf samples showing symptoms of puckering and
yellow or light green mottling were collected from stunted plants from
different locations. Total RNA was extracted from these samples using
TRIzol
®
Reagent (Life Technologies, Carlsbad, USA). Six of the 25
samples were positive by one step RT-PCR using PMMoV-specific primers
MP/for (5′-TAAAATTGGGCAGAACTCGGAG-3′ and 3'UTR/rev
(5′-ACGACAACCCTTCGATTTAAGT-3′), designed from an alignment
of PMMoV sequences. After purification using the Amicon
®
Ultra kit
(EMD Millipore, Billerica, USA) the amplicons were directly sequenced in
both directions (BioFab, Rome, Italy). A phylogenetic tree was obtained
using the neighbour-joining method based on the Kimura 2-parameter
model in MEGA6 (Tamura et al., 2013). The sequences of the 629 bp
amplicon from all six isolates, including the complete coat protein gene
(474 bp), were identical. The sequence of one isolate (AAR-PK) was
submitted to GenBank (Accession No. KT853037). BLASTn analyses
showed >99% sequence identity of the Pothwar-Pakistan isolate of
PMMoV with PMMoV isolates from Asia, including China (KP345899),
India (KJ631123) and Japan (KJ631123). Phylogenetic analysis (Fig. 1)
supported the grouping of the Pothwar-Pakistan isolate within the
pathotype P12 cluster (Caglar et al., 2013; Rialch et al., 2015), although
validation of the pathotype would require phenotyping on differential hosts
including alleles with L
2
gene-mediated resistance.
To the best of our knowledge, this is the first report of PMMoV infecting
chilli pepper in Pakistan. Therefore, there is a need to strictly control seed
quality and to extend surveillance to assess PMMoV incidence in other
chilli-growing areas of Pakistan.
References
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characterization of Pepper mild mottle virus from Turkey. Journal of
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Farooq O, 2014. Agriculture. In: Wasti SE, ed. Economic Survey of
Pakistan 2013-14. Islamabad, Pakistan: Ministry of Finance, Government
of Pakistan, 23-41. Retrieved 25 November 2015 from
http://finance.gov.pk/survey_1314.html
Rialch N, Sharma V, Sharma A, Sharma PN, 2015. Characterization and
complete nucleotide sequencing of Pepper mild mottle virus infecting bell
pepper in India. Phytoparasitica 43, 327-337.
http://dx.doi.org/10.1007/s12600-015-0453-6
King AMQ, Adams MJ, Carstens EB, Lefkowitz EJ, eds, 2012. Virus
Taxonomy: Classification and Nomenclature of Viruses. Ninth Report of the
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Academic Press.
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To cite this report: Ahmad A, Tiberini A, Ashfaq M, Tomassoli L, 2015. First report of Pepper mild mottle virus infecting chilli pepper in
Pakistan. New Disease Reports 32, 31. http://dx.doi.org/10.5197/j.2044-0588.2015.032.031
©2015 The Authors
This report was published on-line at www.ndrs.org.uk where high quality versions of the figures can be found.
New Disease Reports is a peer-reviewed on-line journal published by the British Society for Plant Pathology,
for more information visit http://www.ndrs.org.uk/
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