Virus-induced Gene Silencing in Streptocarpus rexii (Gesneriaceae)

Abstract

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.

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Data Availability

The SrPDS sequence generated and/or analysed during the current study are available in the BLAST repository [NCBI GenBank ID MT127415; https://www.ncbi.nlm.nih.gov].

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Acknowledgements

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.

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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.

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Correspondence to Kanae Nishii or Attila Molnar.

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Nishii, K., Fei, Y., Hudson, A. et al. Virus-induced Gene Silencing in Streptocarpus rexii (Gesneriaceae). Mol Biotechnol 62, 317–325 (2020). https://doi.org/10.1007/s12033-020-00248-w

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Keywords

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