RNAi-Mediated Simultaneous Resistance Against Three RNA Viruses in Potato
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RNA interference (RNAi) technology has been successfully applied in stacking resistance against viruses in numerous crop plants. During RNAi, the production of small interfering RNAs (siRNAs) from template double-standard RNA (dsRNA) derived from expression constructs provides an on-switch for triggering homology-based targeting of cognate viral transcripts, hence generating a pre-programmed immunity in transgenic plants prior to virus infection. In the current study, transgenic potato lines (Solanum tuberosum cv. Desiree) were generated, expressing fused viral coat protein coding sequences from Potato virus X (PVX), Potato virus Y (PVY), and Potato virus S (PVS) as a 600-bp inverted repeat expressed from a constitutive 35S promoter. The expression cassette (designated Ec1/p5941) was designed to generate dsRNAs having a hairpin loop configuration. The transgene insertions were confirmed by glufosinate resistance, gene-specific PCR, and Southern blotting. Regenerated lines were further assayed for resistance to virus inoculation for up to two consecutive crop seasons. Nearly 100% resistance against PVX, PVY, and PVS infection was observed in transgenic lines when compared with untransformed controls, which developed severe viral disease symptoms. These results establish the efficacy of RNAi using the coat protein gene as a potential target for the successful induction of stable antiviral immunity in potatoes.
KeywordsRNAi Interaction Resistance Virus Transgenic Potato PVX PVY PVS
This work was funded by Higher Education Commission (HEC), Government of Pakistan, under the indigenous PhD fellowship and research initiative program (IRSIP), and United States Department of Agriculture award 2014-67013-21659 to GJ. The authors would like to thank Ms. Kerry Swartwood for the assistance in tissue culture work conducted at the Boyce Thomason Institute for Plant Research.
Compliance with Ethical Standards
Conflict of interest
The authors declare no financial or commercial conflict of interest.
- 1.Jeffries, C., Barker, H., & Khurana, S. (2006). Potato viruses (and viroids) and their management. In S. M. Khurana (Ed.), Handbook of Potato production, improvement and post-harvest management (Gopal J. New York: The Haworth’s Food Products Press.Google Scholar
- 2.Chaube, H. S., & Pundhir, V. S. (2005). Crop diseases and their management. New Delhi: PHI Learning Pvt. Ltd.Google Scholar
- 4.Fletcher, J. (2012). A virus survey of New Zealand fresh, process and seed potato crops during 2010–11. New Zealand Plant Protection, 65, 197–203.Google Scholar
- 10.Chung, B. N., Yoon, J. Y., & Palukaitis, P. (2013). Engineered resistance in potato against Potato leafroll virus, Potato virus A and Potato virus Y. Virus Genes, 47, 86–92. Google Scholar
- 12.Lawson, C., Kaniewski, W., Haley, L., Rozman, R., Newell, C., Sanders, P., et al. (1990). Engineering resistance to mixed virus infection in a commercial potato cultivar: Resistance to Potato virus X and Potato virus Y in transgenic Russet Burbank. Nature Biotechnology, 8, 127–134.CrossRefGoogle Scholar
- 19.Saurabh, S., Vidyarthi, A. S., & Prasad, D. (2014). RNA interference: Concept to reality in crop improvement. Planta, 239, 543–564.Google Scholar
- 22.Sambrook, J., Frisch, E. F., & Maniatis, T. (1989). Molecular cloning: A laboratory manual. New York: Cold Spring Harbor Laboratory Press.Google Scholar
- 24.Ausubel, F. M. (2002). Short protocols in molecular biology: A compendium of methods from current protocols in molecular biology. 2. New York: Wiley.Google Scholar
- 25.Khatoon, S., Kumar, A., Sarin, N. B., & Khan, J. A. (2016). RNAi-mediated resistance against Cotton leaf curl disease in elite Indian cotton (Gossypium hirsutum) cultivar Narasimha. Virus Genes, 52, 530–537.Google Scholar