title: RNAi-Mediated Simultaneous Resistance Against Three RNA Viruses in Potato.
abstract: 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.
authors.affiliation: Molecular Virology and Gene Silencing Group, National Institute for Biotechnology and Genetic Engineering, Faisalabad, 38000, Pakistan.|Department of Bioinformatics and Biotechnology, Government College University, Allama Iqbal Road, Faisalabad, Pakistan.
authors.forename: Amir
authors.initials: A
authors.lastname: Hameed
authors.affiliation: Molecular Virology and Gene Silencing Group, National Institute for Biotechnology and Genetic Engineering, Faisalabad, 38000, Pakistan.
authors.forename: Muhammad Nouman
authors.initials: MN
authors.lastname: Tahir
authors.affiliation: Molecular Virology and Gene Silencing Group, National Institute for Biotechnology and Genetic Engineering, Faisalabad, 38000, Pakistan.
authors.forename: Shaheen
authors.initials: S
authors.lastname: Asad
authors.affiliation: Molecular Virology and Gene Silencing Group, National Institute for Biotechnology and Genetic Engineering, Faisalabad, 38000, Pakistan.
authors.forename: Rakhshanda
authors.initials: R
authors.lastname: Bilal
authors.affiliation: Boyce Thompson Institute for Plant Research, 533 Tower Road, Ithaca, NY, 14853, USA.
authors.forename: Joyce
authors.initials: J
authors.lastname: Van Eck
authors.affiliation: Boyce Thompson Institute for Plant Research, 533 Tower Road, Ithaca, NY, 14853, USA.
authors.forename: Georg
authors.initials: G
authors.lastname: Jander
authors.affiliation: Molecular Virology and Gene Silencing Group, National Institute for Biotechnology and Genetic Engineering, Faisalabad, 38000, Pakistan. shahidmansoor7@gmail.com.
authors.forename: Shahid
authors.initials: S
authors.lastname: Mansoor
mesh_terms: D036022:Capsid Proteins / Q000037:antagonists & inhibitors / Q000235:genetics; D060467:Disease Resistance; D055029:Inverted Repeat Sequences; D010935:Plant Diseases / Q000235:genetics; D030821:Plants, Genetically Modified; D017800:Potyvirus / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D034622:RNA Interference; D011198:Solanum tuberosum / Q000235:genetics* / Q000821:virology
chemical_list: D036022:Capsid Proteins
keywords: Interaction; PVS; PVX; PVY; Potato; RNAi; Resistance; Transgenic; Virus