European Journal of Plant Pathology

, Volume 150, Issue 1, pp 205–212 | Cite as

Field survey of ranunculus mild mosaic virus, tomato spotted wilt virus and cucumber mosaic virus infections in Ranunculus asiaticus L. in Japan by newly developed multiplex RT-PCR

  • Saki Hayahi
  • Yosuke Matsushita
  • Yoshiaki Kanno
  • Yoshiyuki Kushima
  • Satoshi Teramoto
  • Minoru TakeshitaEmail author


Viral symptoms are frequently observed in production fields of ranunculus (Ranunculus asiaticus L.) in Japan. Based on incidence of diseases caused by a large number of ranunculus-infective viruses, ranunculus mild mosaic virus (RanMMV), tomato spotted wilt virus (TSWV) and cucumber mosaic virus (CMV) infections were the focus of an epidemiological field survey in Japan. To efficiently investigate the incidence and distribution of the three viruses, we first developed a new multiplex reverse transcription-polymerase chain reaction method that enables simultaneous detection of RanMMV, TSWV, and CMV in ranunculus. A field survey of virus infections in ranunculus production fields in Tohoku and Kyushu regions revealed that the infection rate of RanMMV was much higher than that of the other viruses in all fields sampled. Interestingly, the infection rate of RanMMV showed an increasing trend in proportion to the number of vegetative propagation cycles of ranunculus, implying virus transmission by aphids. Taken together, this method was proven to be effective for simultaneous detection of RanMMV, TSWV and CMV in ranunculus plants, and RanMMV was recognized as one of the most prevalent plant viruses in ranunculus fields in Japan.


Ranunculus asiaticus L. Ranunculus mild mosaic virus Tomato spotted wilt virus Cucumber mosaic virus 



We thank Dr. K. Nakamura for critical reading of this manuscript, and Mr. Y. Yoshida, Ms. E. Kodama, Ms. T. Kitamura, and Ms. M. Nagatomo for preparing the experimental materials. This study was supported by a Grant-in-Aid for “Research and development projects for application in promoting new policy of agriculture, forestry and fisheries (25075c)” from the Ministry of Agriculture, Forestry, and Fisheries of Japan.

Compliance with ethical standards

Conflict of interest



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Copyright information

© Koninklijke Nederlandse Planteziektenkundige Vereniging 2017

Authors and Affiliations

  • Saki Hayahi
    • 1
    • 2
  • Yosuke Matsushita
    • 3
  • Yoshiaki Kanno
    • 4
  • Yoshiyuki Kushima
    • 1
  • Satoshi Teramoto
    • 1
  • Minoru Takeshita
    • 5
    Email author
  1. 1.Miyazaki Agricultural Research InstituteMiyazakiJapan
  2. 2.Nishiusuki Branch Office Miyazaki Prefectural GovernmentMiyazakiJapan
  3. 3.Institute of Vegetable and Floriculture Science, NAROIbarakiJapan
  4. 4.Minami Kyushu UniversityMiyazakiJapan
  5. 5.University of MiyazakiMiyazakiJapan

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