New molecular diagnostic methods for detection of Chrysanthemum stem necrosis virus (CSNV)


  • Jana Boben Agricultural institute of Slovenia, Hacquetova 17, Ljubljana, Slovenia
  • Nataša Mehle
  • Manca Pirc
  • Irena Mavrič Pleško
  • Maja Ravnikar



CSNV, diagnostics, distinguishing, ELISA, real-time PCR, test plants, tospoviruses, TSWV, RT-PCR


Chrysanthemum stem necrosis virus (CSNV), an RNA virus, belongs to the genus Tospovirus and family Bunyaviridae. The disease symptoms on host plants can not be distinguished from those caused by closely related viruses of the same genus, such as Tomato spotted wilt virus (TSWV). The disease symptoms may vary between host plants and can be quite severe. Diagnostics of tospoviruses is of great importance in prevention of greater economic and environmental damage. Viruses within the genus show serological similarity and therefore make the detection of the viruses by serological methods unreliable. To avoid false positive results other methods can be used for detection of RNA viruses such as RT-PCR or RT real-time PCR. CSNV was first found in Slovenia in 2001. Different methods were used to confirm the identity of the isolate. Development of disease symptoms was observed on 15 different test plants and was compared to the symptoms caused by closely related TSWV. Since serological cross-reac- tivity was observed, molecular tests were developed (RT-PCR and real-time PCR) that further confirmed the identity of the isolate and also increased sensitivity of the assays. CSNV can now be reliably detected and distinguished from related tospoviruses.


ALEXANDRE M. A. V., E. B.RIVAS, L. M DUARTE. & M. H. OKUYAMA 1996: Serological survey of tospo- virus on Chrysanthemum sp. Crops in São Paulo state, Brazil. Fitopatol. Bras. 21 (1): 80–84.

Acta Biologica Slovenica, 50 (1), 2007

ALEXANDRE M. A. V., L. M. L. DUARTE, E. B. RIVAS & C. M. CHAGAS 1999: Mixed infection by Tospo- virus species in ornamental crops in Sao Paulo state, Brazil. Summa phytopathol. 25: 353–356.

BEZERRA I. C., R. DE O. RESENDE, L. POZZER, T. NAGATA, R. KORMELINK & A. C. DE ÁVILA 1999: Increase of tospoviral diversity in Brazil with the identification of two new tospovirus species, one from chrysanthemum and one from zucchini. Phytopathology 89 (9): 823–830. DOI:

BOONHAM N., P. SMITH, K. WALSH, J. TAME, J. MORRIS, N. SPENCE, J. BENNISON & I. BARKER 2002: The detection of Tomato spotted wilt virus (TSWV) in individual thrips using real time fluorescent RT-PCR (TaqMan). J. Virol. Methods 101: 37–48. DOI:

CORTEZ I., J. SAAIJER, K. S. WONGJKAEW, A.-M. PEREIRA, R. GOLDBACH, D. PETERS & R. KORMELINK 2001: Identification of a novel tospovirus species using a new RT-PCR approach. Arch. Virol. 146: 265–278. DOI:

DUARTE L. M. L., E. B. RIVAS, M. A. V. ALEXANDRE, A. C. DE ÁVILA, T. NAGATA & C. M. CHAGAS 1995: Chrysanthemum stem necrosis caused by a possible novel Tospovirus. J. Phytopathol. 143: 569–571. DOI:

EIRAS M., R. O. RESENDE, A. A. MISSIAGGIA & A. C. DE ÁVILA 2001: RT-PCR and dot blot hybridization methods for a universal detection of tospoviruses. Fitopatol. Bras. 26 (2): 170–175. DOI:

NICHOL S. T., B. J. BEATY, R. M. ELLIOT, R. GOLDBACH, A. PLYUSNIN, C. S. SCHMALJOHN & R. B. TESH 2005: GENUS TOSPOVIRUS. IN: FAQUET C. M., M. A. MAYO, J. MANILOFF, U. DESSELBERGER & L. A. BALL (EDS.): Virus Taxonomy. Classification and Nomenclature of Viruses, Eighth Report of the International Committee on Taxonomy of Viruses. Elsevier Academic Press, Amsterdam, pp. 712–716.

DE HAAN P., L. WAGEMAKERS, D. PETERS & R. GOLDBACH 1990: The S RNA segment of tomato spotted wilt virus has an ambisense character. J. Gen. Virol. 71: 1001–1007. DOI:

HEINZE C., B. LETSCHERT, D. HRISTOVA, M. YANKULOVA, O. KAUADJOUOR, P. WILLINGMANN, A. ATANASSOV & G. ADAM 2001: Variability of the N-protein and the intergenic region of the S RNA of Tomato spotted wilt tospovirus (TSWV). Microbiologica 24: 175–187.

LETSCHERT B., G. ADAM, D.-E. LESSEMAN, P. WILLINGMANN & C. HEINZE 2002: Detection and differen- tiation of serologically cross-reacting tobamoviruses of economical importance by RT-PCR and RT-PCR-RFLP. J. Virol. Methods 106: 1-10. DOI:

MATSUURA S., K. KUBOTA & M. OKUDA 2007: First Report of Chrysanthemum stem necrosis virus on Chrysanthemums in Japan. Plant Dis. 91 (4): 468. DOI:

MAVRIČ I. & M. RAVNIKAR 2001: First report of Tomato spotted wilt virus and Impatiens necrotic spot virus in Slovenia. Plant Dis. 85 (12): 1288. DOI:

MUMFORD R. A., I. BARKER & K. R. WOOD 1994: The detection of tomato spotted wilt virus using the polymerase chain reaction. J. Virol. Methods 46: 303–311. DOI:

MUMFORD R. A., B. JARVIS, J. MORRIS & A. BLOCKLEEY 2003: The first report of Chrysanthemum stem necrosis virus in United Kingdom. New Disease Reports, 7:feb.2003–jul.2003, http://www.bspp. (dec. 2005).

NAGATA T., R. DE O. RESENDE, E. W. KITAJIMA, H. COSTA, A. K. INOUE-NAGATA & A. C. DE ÁVILA 1998: First report of natural occurence of Zucchini lethal chlorosis tospovirus on cucumber and Chry- santhemum stem necrosis tospovirus on tomato in Brazil. Plant Dis. 82: 1403. DOI:

NAGATA T. & A. C. DE ÁVILA 2000: Transmission of Chrysanthemum Stem Necrosis Virus, a Recently Discovered Tospovirus, by Two Thrips Species. J. Phytopathol. 148 (2): 123–125. DOI:

OKUDA M. & K.HANADA 2001: RT-PCR for detecting five distinct Tospovirus species using degenerate primers and dsRNA template. J. Virol. Methods 96: 149–156. DOI:

PAPPU S. S., A. I. BHAT, H. R. PAPPU, C. M. DEOM & A. K. CULBREATH 2000: Phylogenetic studies of tospoviruses (Family: Bunyaviridae) based on intergenic region sequences of small and medium genomic RNAs. Arch. Virol. 145: 1035–1045. DOI:

RESENDE R. DE O., L. POZZER, T. NAGATA, I. C. BEZERRA, M. I. LIMA, L. B. GIORDANO, E. W. KITAJIMA & A. C. DE ÁVILA 1996: New tospoviruses found in Brazil. Acta Horticulturae 431: 78–81. DOI:

J. Boben, N. Mehle, M. Pirc, I. Mavrič Pleško, M. Ravnikar: New molecular diagnostic methods ...

ROBERTS C. A., R. G. DIETZGEN, L. A. HEELAN & D. J. MACLEAN 2000: Real-time RT-PCR fluorescent detection of tomato spotted wilt virus. J. Virol. Methods 88: 1–-8. DOI:

SALMON M. A., M. VENDRAME, J. KUMMERT & P. LEPOIVRE 2002: Detection of apple chlorotic leaf spot virus using a 5’ nuclease assay with a fluorescent 3’ minor groove binder-DNA probe. J. Virol. Methods 104: 99–106. DOI:

UGA H. & S. TSUDA 2005: A one-step reverse transcription-polymerase chain reaction system for the simultaneous detection and identifiaction of multiple tospovirus infections. Phytopathology 95: 166–171. DOI:

VERHOEVEN J. TH. J., J. W. ROENHORST, I. CORTES & D. PETERS 1996: Detection of a novel tospovirus in Chrysanthemum. Acta Horticulturae 432: 44–51. DOI:

WELLER S. A., J. G. ELPHINSTONE, N. C. SMITH, N. BOONHAM & D. E. STEAD 2000: Detection of Ralstonia solanacearum strains with a quantitative, multiplex, real-time, fluorogenic PCR (TaqMan) assay. Appl. Environ. Microb. 66 (7): 2853–2858. DOI:

WILLIAMS L. V., P. M. LOPEZ LAMBERTINI, K. SHOHARA & E. B. BIDERPOST 2001: Occurence and geo- graphical distribution of tospocirus species infecting tomato crops in Argentina. Plant Dis. 85: 1227–1229. DOI:






Original Research Paper

How to Cite

Boben, J., Mehle, N., Pirc, M., Mavrič Pleško, I., & Ravnikar, M. (2007). New molecular diagnostic methods for detection of Chrysanthemum stem necrosis virus (CSNV). Acta Biologica Slovenica, 50(1), 41-51.

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