Plant Pathology (2005) 54, 807
NEW DISEASE REPORT
Blackwell Publishing, Ltd.
First report of Tomato leaf curl New Delhi virus infecting
bitter gourd in Pakistan
M. Tahir and M. S. Haider*
School of Biological Sciences, University of the Punjab, Quaid-i-Azam Campus, Lahore, Pakistan
Bitter gourd (Momordica charantia; family Cucurbitaceae)
is a vegetable cultivated in many areas of Pakistan. Plants
showing yellow blotch symptoms were observed in
several fields in the vicinity of Lahore, Pakistan, with an
average incidence of 60 –70%. Leaf samples were collected
from four diseased and from two apparently healthy
(symptomless) plants, and used for DNA extraction.
The presence of a begomovirus was confirmed by PCR
amplification using a degenerate primer pair designed to
conserved regions of the coat protein genes from published
sequences of begomoviruses from the Old World (Haider,
1996; virion-sense primer 5′-ATG(C/A/T)(G/C) (G/C/
A)AAGCG(A/ T)(C/A)C(A /C)G(G/C)(A /C)GATAT-3′;
complementary-sense primer 5′-TTAATT(T/G/C)(C/G/
A)(A/ T/C)(A/ T/G)A(C/T)(A/T/C)(G/C)(C/A/T)(A/G)
TCATA(G/A)AA(A /G)TA-3′). An amplification product of
the expected size (≈ 750 bp) was produced from samples
from plants with symptoms, but not from symptomless
plants. The PCR product was cloned and sequenced
(accession no. AJ854186). The sequence showed the highest
levels of sequence identity (95%) to Tomato leaf curl
New Delhi virus (ToLCNDV; U15015), indicating that the
virus of M. charantia is a strain of ToLCNDV.
Recent reports indicate that many Old World begomo-
viruses are associated with a single-stranded DNA satellite
(DNA-β; Briddon et al., 2003). Attempts to identify the
*E-mail: haider65us@yahoo.com
Accepted 31 January 2005 at www.bspp.org.uk/ndr where
figures relating to this paper can be viewed
© 2005 BSPP
Doi: 10.1111/j.1365-3059.2005.01215.
presence of a DNA-β in infected M. charantia using uni-
versal DNA-β primers (Briddon et al., 2002) produced a
0·6 kb band that may be a defective DNA-β. Efforts are
now under way to sequence this and to produce full-length
clone(s) of the virus. This is the first report of ToLCNDV
infecting M. charantia.
Acknowledgements
The authors would like to thank Dr Rob Briddon (Higher
Education Commission [Pakistan] – Foreign Faculty Hiring
Programme) for assistance in preparing the manuscript.
References
Briddon RW, Bull SE, Mansoor S, Amin I, Markham PG,
2002. Universal primers for the PCR-mediated amplification
of DNA-β; a molecule associated with some monopartite
begomoviruses. Molecular Biotechnology 20, 315 –8.
Briddon RW, Bull SE, Amin I, Idris AM, Mansoor S, Bedford ID,
Dhawan P, Rishi N, Siwatch SS, Abdel-Salam AM, Brown JK,
Zafar Y, Markham PG, 2003. Diversity of DNA; a satellite
molecule associated with some monopartite begomoviruses.
Virology 312, 106 –21.
Haider MS, 1996. Characterization of Whitefly-Transmitted
Geminiviruses from Pakistan. PhD thesis, University of
London, UK.
807