Journal of General Plant Pathology

, Volume 76, Issue 4, pp 273–277 | Cite as

Improved multiplex reverse transcription-polymerase chain reaction to detect and identify five tospovirus species simultaneously

  • Katsuya Kuwabara
  • Naoto Yokoi
  • Takehiro Ohki
  • Shinya Tsuda
Viral and Viroid Diseases
First Online:
Received:
Accepted:

Abstract

We improved a one-step multiplex reverse transcription-polymerase chain reaction (multi-PCR) method previously published for the simultaneous identification of five tospovirus species that mainly occur on Solanaceae and Compositae plants. The multi-PCR system is composed of a universal degenerate primer and five virus-species-specific primers that amplify bands unique to Iris yellow spot virus (837 bp), Tomato spotted wilt virus (709 bp), Impatiens necrotic spot virus (589 bp), Chrysanthemum stem necrosis virus (485 bp), and Capsicum chlorosis virus (366 bp). This technique is potentially useful for epidemiological studies of several tospoviruses that have recently been detected in previously uninfected fields of ornamental crops.

Keywords

Capsicum chlorosis virus Chrysanthemum stem necrosis virus Degenerate primer Diagnosis Epidemiology Tospovirus 
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Notes

Acknowledgments

We are grateful to L. M. Knight for critically reading this manuscript. We thank Y. Matsumura and S. Nagai for the preparing and maintaining the plants.

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Copyright information

© The Phytopathological Society of Japan and Springer 2010

Authors and Affiliations

  • Katsuya Kuwabara
    • 1
  • Naoto Yokoi
    • 2
  • Takehiro Ohki
    • 3
  • Shinya Tsuda
    • 3
  1. 1.Gunma Agricultural Technology CenterIsesakiJapan
  2. 2.Akita Agricultural Extension Office at Yamamoto RegionNoshiroJapan
  3. 3.National Agricultural Research CenterTsukubaJapan

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