African Journal of
Microbiology Research

  • Abbreviation: Afr. J. Microbiol. Res.
  • Language: English
  • ISSN: 1996-0808
  • DOI: 10.5897/AJMR
  • Start Year: 2007
  • Published Articles: 4970

Full Length Research Paper

Multiplex RT-PCR identification of five viruses associated with the watermelon crops in the Brazilian Cerrado

Raimundo Wagner de Souza Aguiar
  • Raimundo Wagner de Souza Aguiar
  • Plant Production Graduate Program, Federal University of Tocantins, Gurupi-TO, Brazil.
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Adelmo Rodrigues Martins
  • Adelmo Rodrigues Martins
  • Plant Production Graduate Program, Federal University of Tocantins, Gurupi-TO, Brazil.
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Vitor L. Nascimento
  • Vitor L. Nascimento
  • Plant Production Graduate Program, Federal University of Tocantins, Gurupi-TO, Brazil.
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Aristóteles Capone
  • Aristóteles Capone
  • Plant Production Graduate Program, Federal University of Tocantins, Gurupi-TO, Brazil.
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Lara Teixeira Melo Costa
  • Lara Teixeira Melo Costa
  • Departament of Biotechnology, Universidade Federal do Tocantins, Gurupi, TO, 77413-070, Brazil.
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Fabrício Souza Campos
  • Fabrício Souza Campos
  • Departament of Biotechnology, Universidade Federal do Tocantins, Gurupi, TO, 77413-070, Brazil.
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Rodrigo Ribeiro Fidelis
  • Rodrigo Ribeiro Fidelis
  • Plant Production Graduate Program, Federal University of Tocantins, Gurupi-TO, Brazil.
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Gil Rodrigues dos Santos
  • Gil Rodrigues dos Santos
  • Plant Production Graduate Program, Federal University of Tocantins, Gurupi-TO, Brazil.
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Renato de Oliveira Resende
  • Renato de Oliveira Resende
  • Department of Cell Biology, Universidade de Brasilia, Brasilia-DF, Brazil.
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Tatsuya Nagata
  • Tatsuya Nagata
  • Department of Cell Biology, Universidade de Brasilia, Brasilia-DF, Brazil.
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  •  Received: 03 September 2018
  •  Accepted: 13 December 2018
  •  Published: 21 January 2019

Abstract

Watermelon plants can be naturally infected by several viruses in single or mixed infections, of which the diagnosis is difficult and require specific techniques. This study aims to detect and to verify the presence of Cucumber mosaic virus (CMV), Papaya ringspot virus type W (PRSV-W), Watermelon mosaic virus (WMV), Zucchini lethal chlorosis virus (ZLCV), Zucchini yellow mosaic virus (ZYMV), and the main cucurbit viruses in Brazil using multiplex reverse transcriptase polymerase chain reaction (RT-PCR) assay. Oligonucleotides were designed towards the conserved regions of the virus genomes. In the duplex-PCR, it was possible to detect all the virus combinations, except ZLCV with ZYMV. The amplified product sizes were 644 bp (CMV), 535 bp (WMV-2), 398 bp (PRSV-W), 244 bp (ZLCV), and 214 bp (ZYMV). To test the efficacy of the methodology, we analyzed plants with virus symptoms from four municipalities in the state of Tocantins, located at Brazilian Cerrado. Mixed infections were detected in 80% of samples in all the municipalities. The multiplex RT-PCR assay can be used to detect and differentiate watermelon viruses that affect crops in the state of Tocantins.

 

Key words: Cucumovirus, multiplex RT-PCR, Orthotospovirus, potyvirus, watermelon.