DISEASE NOTE
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FIRST REPORT OF EUPHORBIA YELLOW LEAF CURL VIRUS INFECTING HIBISCUS SYRIACUS H. Riaz, M. Ashfaq, T. Mukhtar and T. Riaz
Department of Plant Pathology, Pir Mehr Ali Shah Arid Agriculture University, Rawalpindi, Pakistan
Hibiscus syriacus is a flowering plant of the family Malvaceae known as a host of
begomoviruses around the world. One plant showing symptoms of leaf curling and yellowing was collected along with a symptomless one from which total DNA was extracted using the
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CTAB method. Initial begomovirus detection amplified a 570 bp product from the symptomatic plant identified as the viral coat protein (Wyatt and Brown, 1996), which was followed by rolling
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circle amplification (RCA) using random hexamer primers (Inoue-Nagata et al., 2004). The RCA product was digested with BamHI, yielding a ca. 2.8 kb DNA fragment that was subsequently cloned into pCambia1301. The sequence of the complete genome of a viral isolate denoted PK2 was deposited in GenBank under the accession No. KP780424. No associated genomic
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components (DNA-B/DNA β) were detected using the BC1F/BC1R (Hussain et al., 2004) and Beta01/Beta02 (Briddon et al., 2002) primer pairs. The isolate PK2 proved to be DNA-A (2,728
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bp) with the typical old world begomovirus genomic orientation. Identity scores were calculated by SDT, which showed that isolate PK2 had maximum 98.8% pairwise identity with Euphorbia
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yellow leaf curl virus (EYLCuV) (accession No. KM978186). According to the latest demarcation threshold of 91%, PK2 was identified as an isolate of Euphorbia yellow leaf curl virus (KM978186) (Brown et al., 2015). To the best of our knowledge it is the first report of
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EYLCuV infecting a member of the family Malvaceae.
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The authors are thankful to Higher Education Commission of Pakistan for providing funding for the project under the HEC Indigenous PhD fellowship program.
Briddon R.W., Bull S.E., Mansoor S., Amin I., Markham P.G., 2002. Universal primers for the PCR mediated amplification of DNA β; a molecule associated with some monopartite begomoviruses. Molecular Biotechnology 20: 315-318.
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Brown J.K., Zerbini F.M., Navas-Castillo J., Moriones E., Ramos-Sobrinho R., Silva J.C.F., FialloOlive E., Briddon R.W., Herna´ndez-epeda C., Idris A., Malathi V.G., Martin D.P., RiveraBustamante R., Ueda S., Varsani A., 2015. Revision of Begomovirus taxonomy based on pairwise sequence comparisons. Archives of Virology 160:1593-1619. Hussain M., Mansoor S., Iram S., Zafar Y., Briddon R.W., 2004. First report of Tomato leaf curl New Delhi virus affecting chilli pepper in Pakistan. Plant Pathology 53: 794.
Inoue-Nagata A.K., Albuquerque L.C., Rocha W.B., Nagata T., 2004. A simple method for cloning the complete begomovirus genome using the bacteriophage φ 29 DNA polymerase. Journal of Virological Methods 116: 209-211. Wyatt S.D., Brown J.K., 1996. Detection of Subgroup III Geminivirus isolates in leaf extracts by degenerate primers and polymerase chain reaction. Phytopathology 86: 1288-1293.
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Received June 17, 2015 Accepted September 17, 2015
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Corresponding author: M. Ashfaq E-mail:
[email protected]
