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First report of tomato spotted wilt virus on lisianthus (Eustoma grandiflorum) in Bulgaria

Tomato spotted wilt virus (TSWV) is one of the most destructive pathogens worldwide. In Bulgaria the virus was found to severely damage a number of solanaceous vegetables and ca. 25 ornamentals (Hristova et al. 2001). In spring 2014, about 30% of lisianthus plants (ca. 1000 plants) grown in a private farm nearby Plovdiv, Bulgaria showed yellowing and/or light brown necrosis on leaves and petals followed by subsequent wilting. To identify the causal agent of this infection, Chenopodium quinoa, Nicotiana glutinosa, and Petunia hybrida plants were sap inoculated with material from 10 symptomatic individuals. C. quinoa displayed large necrotic local lesions. Primary and systemic symptoms in N. glutinosa appeared as translucent necrotic lesions accompanied with yellow areas, while P. hybrida showed dark necrotic ring spots. Observations of lisianthus leaf sap by electron microscopy revealed the presence of isometric particles ca. 85 nm in size resembling those of tospoviruses. A TAS-ELISA test was conducted with antisera specific for impatiens necrotic spot virus (INSV, RT-0115-0117/1) and TSWV (RT-0105-0106/3) purchased from DSMZ, Germany. The samples from lisianthus and those from the inoculated indicators were positive only for TSWV. RNA was isolated from symptomatic leaves of lisianthus by TRI reagent® (Sigma-Aldrich) and reverse transcription (RT) was performed using random hexamer primers. The PCR amplification conducted with specific primers targeting N gene, described by Roberts et al. (2000), revealed an amplicon of about 600 bp corresponding to the expected size. Further, the amplicon was sent for sequencing. A high-confidence consensus nucleotide sequence of 567 bp (accession No. MK388405) was selected for analysis, which encoded a protein of 188 amino acids. This protein was queried against a database of all NCBI viral protein sequences by blastp and showed 99% sequence identity to TSWV nucleocapsid protein, including several accessions from Bulgaria (e.g. CAC01282.1, CAC01285.1) (Heinze et al. 2001). The results from TAS-ELISA and RT-PCR tests clearly show that the virus-specific antibodies do not have interspecies cross-reactivity. Based on our knowledge and viruses reported in literature, this appears to be a first report of TSWV on lisianthus in Bulgaria.

References

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  • Hristova D, Karadjova O, Yankulova M, Heinze C, Adam G (2001) A survey of tospoviruses in Bulgaria. J Phytopathol 149:745–749

  • Roberts CA, Dietzgen R, Heelan L, Maclean D (2000) Real-time RT-PCR fluorescent detection of tomato spotted wilt virus. J Virol Methods 88:1–8

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Acknowledgements

The authors GP, VR-I, RL, ANN, and DK acknowledge the financial support of Horizon 2020 PlantaSYST project under the Grant Agreement No 739582.

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Correspondence to Gancho Pasev.

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Pasev, G., Radeva-Ivanova, V., Lyall, R. et al. First report of tomato spotted wilt virus on lisianthus (Eustoma grandiflorum) in Bulgaria. J Plant Pathol 103, 375 (2021). https://doi.org/10.1007/s42161-020-00702-3

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  • DOI: https://doi.org/10.1007/s42161-020-00702-3

Keywords

  • TSWV
  • Lisianthus
  • ELISA
  • RT-PCR
  • Bulgaria