Virus-induced Gene Silencing in Streptocarpus rexii (Gesneriaceae)


Many members of the family Gesneriaceae are cultivated as ornamental plants, including Cape primrose (Streptocarpus) species. The range of plant architecture found in this genus has also made it a model to study leaf and meristem development and their evolution. However, the lack of tools to study gene functions through reverse genetics in Streptocarpus has limited the exploitation of its genetic potential. To aid functional genomic studies in Streptocarpus rexii, we sought to investigate virus-induced gene silencing (VIGS). Using the broad host range Tobacco Rattle Virus (TRV) to target the PHYTOENE DESATURASE (PDS) gene of S. rexii, we show that infection with sap from Nicotiana benthamiana triggered VIGS efficiently. VIGS was most effective in the seedling leaves 8 weeks after sowing, but was limited in duration and systemic spread. This study reports the first successful use of VIGS in Streptocarpus and in the family Gesneriaceae. The inoculation of viral sap derived from N. benthamiana was able to overcome the difficulties of standard Agrobacterium-mediated transformation in this genus. Irrespective of its transient effect, this VIGS system will be useful to assess gene function at the cellular level and represent an important tool for further understanding molecular mechanisms in Streptocarpus.

This is a preview of subscription content, access via your institution.

Fig. 1
Fig. 2
Fig. 3

Data Availability

The SrPDS sequence generated and/or analysed during the current study are available in the BLAST repository [NCBI GenBank ID MT127415;].


  1. 1.

    Jong, K., & Burtt, B. L. (1975). The evolution of morphological novelty exemplified in the growth patterns of some Gesneriaceae. New Phytologist,75, 297–311.

    Article  Google Scholar 

  2. 2.

    Burtt, B. L. (1963). Studies in the Gesneriaceae of the Old World, XXIV: tentative keys to the tribes and genera. Notes from the Royal Botanic Garden, Edinburgh,24, 205–220.

    Google Scholar 

  3. 3.

    Nishii, K., Kuwabara, A., & Nagata, T. (2004). Characterization of anisocotylous leaf formation in Streptocarpus wendlandii (Gesneriaceae): Significance of plant growth regulators. Annals of Botany,94, 457–467.

    CAS  Article  Google Scholar 

  4. 4.

    Nishii, K., Huang, B.-H., Wang, C.-N., & Möller, M. (2017). From shoot to leaf: step-wise shifts in meristem and KNOX1 activity correlate with the evolution of a unifoliate body plan in Gesneriaceae. Development Genes and Evolution,227, 41–60.

    Article  Google Scholar 

  5. 5.

    Chiara, M., Horner, D. S., & Spada, A. (2013). De Novo assembly of the transcriptome of the non-model plant Streptocarpus rexii employing a novel heuristic to recover locus-specific transcript clusters. PLoS ONE,8, e80961.

    Article  Google Scholar 

  6. 6.

    Chen, Y.-Y., Nishii, K., Barber, S., Hackett, C., Kidner, C. A., Gharbi, K., et al. (2018). A first genetic map in the genus Streptocarpus generated with RAD sequencing based SNP markers. South African Journal of Botany,117, 158–168.

    Article  Google Scholar 

  7. 7.

    Lange, M., Yellina, A. L., Orashakova, S., & Becker, A. (2013). Virus-induced gene silencing (VIGS) in plants: an overview of target species and the virus-derived vector systems. In A. Becker (Ed.), Virus-induced gene silencing: Methods and protocols, methods in molecular biology (Vol. 975). New York: Springer.

    Google Scholar 

  8. 8.

    Baulcombe, D. C. (1999). Fast forward genetics based on virus-induced gene silencing. Current Opinion in Plant Biology,2, 109–113.

    CAS  Article  Google Scholar 

  9. 9.

    Dinesh-Kumar, S. P., Anandalakshmi, R., Marathe, R., Schiff, M., & Liu, Y. (2003). Virus-induced gene silencing. Methods in Molecular Biology,236, 287–294.

    CAS  PubMed  Google Scholar 

  10. 10.

    Lu, R., Martin-Hernandez, A. M., Peart, J. R., Malcuit, I., & Baulcombe, D. C. (2003). Virus-induced gene silencing in plants. Methods,30, 296–303.

    CAS  Article  Google Scholar 

  11. 11.

    Cheng, C., Gao, J., & Ma, N. (2018). Investigation of petal senescence by TRV-mediated virus-induced gene silencing in rose. Methods in Molecular Biology,1744, 49–63.

    CAS  Article  Google Scholar 

  12. 12.

    Baulcombe, D. (2015). VIGS, HIGS and FIGS: small RNA silencing in the interactions of viruses or filamentous organisms with their plant hosts. Current Opinion in Plant Biology,26, 141–146.

    CAS  Article  Google Scholar 

  13. 13.

    MacFarlane, S. A. (1999). Molecular biology of the tobraviruses. Journal of General Virology,80, 2799–2807.

    CAS  Article  Google Scholar 

  14. 14.

    Ratcliff, F., Martin-Hernandez, A. M., & Baulcombe, D. C. (2001). Technical Advance. Tobacco rattle virus as a vector for analysis of gene function by silencing. The Plant Journal,25, 237–245.

    CAS  Article  Google Scholar 

  15. 15.

    Senthil-Kumar, M., & Mysore, K. S. (2010). Assessing functional role of three water deficit stress-induced genes in nonhost disease resistance using virus-induced gene silencing in Nicotiana benthamiana. Plant Signaling & Behavior,5, 586–590.

    CAS  Article  Google Scholar 

  16. 16.

    Senthil-Kumar, M., & Mysore, K. S. (2014). Tobacco rattle virus-based virus-induced gene silencing in Nicotiana benthamiana. Nature Protocols,9, 1549–1562.

    CAS  Article  Google Scholar 

  17. 17.

    Liu, E., & Page, J. E. (2008). Optimized cDNA libraries for virus-induced gene silencing (VIGS) using tobacco rattle virus. Plant Methods,4, 5.

    Article  Google Scholar 

  18. 18.

    Hughes, M., Möller, M., Bellstedt, D. T., Edwards, T. J., & De Villiers, M. (2005). Refugia, dispersal and divergence in a forest archipelago: A study of Streptocarpus in eastern South Africa. Molecular Ecology,14, 4415–4426.

    CAS  Article  Google Scholar 

  19. 19.

    SCOR-UNESCO. (1996). Determination of photosynthetic pigments in seawater (Vol. 1). UNESCO, Paris: Monographs on Oceanographic Methodology.

    Google Scholar 

  20. 20.

    Pfaffl, M. W., Horgan, G. W., & Dempfle, L. (2002). Relative Expression Software Tool (REST) for group-wise comparison and statistical analysis of relative expression results in Real-Time PCR. Nucleic Acids Research,30, E36.

    Article  Google Scholar 

  21. 21.

    Harries, P., & Ding, B. (2011). Cellular factors in plant virus movement: At the leading edge of macromolecular trafficking in plants. Virology,411, 237–243.

    CAS  Article  Google Scholar 

  22. 22.

    Pyott, D. E., & Molnar, A. (2015). Going mobile: non-cell-autonomous small RNAs shape the genetic landscape of plants. Plant Biotechnology Journal,13, 306–318.

    CAS  Article  Google Scholar 

  23. 23.

    Burch-Smith, T. M., Schiff, M., Liu, Y., & Dinesh-Kumar, S. P. (2006). Efficient virus-induced gene silencing in Arabidopsis. Plant Physiology,142, 21–27.

    CAS  Article  Google Scholar 

  24. 24.

    Kushikawa, S., Hoshino, Y., & Mii, M. (2001). Agrobacterium-mediated transformation of Saintpaulia ionantha Wendl. Plant Science,161, 953–960.

    CAS  Article  Google Scholar 

Download references


We are grateful for the support to KN by P. Hollingsworth at Royal Botanic Garden Edinburgh (RBGE, UK), and A. Iwamoto at Kanagawa University (Japan) and H. Iida at Tokyo Gakugei University (Japan). This work was supported by RBGE's science and horticultural divisions. In specific, we thank M. Hart, F. Christie, R. Holland, and L. Forrest for technical support at RBGE, and S. Barber, N. Kelso and A. Ensoll for their support in cultivating the research materials. This work was financially supported by the Edinburgh Botanic Garden (Sibbald) Trust [Sibbald Trust Grant Number 2017#17] and the Japan Society for the Promotion of Science [JSPS KAKENHI Grant Number 15K18593; 18K06375]. YF was funded by the China Scholarship Council. AM is a Chancellor’s Fellow at the University of Edinburgh. RBGE is supported by the Rural and Environment Science and Analytical Services Division (RESAS) in the Scottish Government.

Author information




All authors contributed to the study conception and design, and manuscript preparation. Data collection and analyses were performed by Yue Fei and Kanae Nishii, with supervision of Attila Molnar, Andrew Hudson, and Michael Möller. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Kanae Nishii or Attila Molnar.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Nishii, K., Fei, Y., Hudson, A. et al. Virus-induced Gene Silencing in Streptocarpus rexii (Gesneriaceae). Mol Biotechnol 62, 317–325 (2020).

Download citation


  • Gesneriaceae
  • Nicotiana benthamiana
  • Streptocarpus rexii
  • Tobacco rattle virus
  • Virus-induced gene silencing