Plant germplasm resistant to RNA viruses
Inventors
Fellers, John • Trick, Harold N. • Cruz, Luisa • Rupp, Jessica
Assignees
Kansas State University • US Department of Agriculture USDA
Publication Number
US-9909139-B2
Publication Date
2018-03-06
Expiration Date
2034-09-24
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Abstract
Disclosed is a dsRNA construct used to silencing specific eukaryotic translation initiation factor in plants to produce a plant resistant to viruses such as Potyviruses, Luteoviruses, and Furoviruses. More specifically, the plant would be resistant to viruses such as Wheat streak mosaic virus, Triticum mosaic virus, Soil bourne mosaic virus, or Barley yellow dwarf virus. Also disclosed are non-transgenic wheat plants having the genes for eIF(iso)4E-2 or eIF4G silenced.
Core Innovation
The invention relates to silencing specific eukaryotic translation initiation factors in plants to produce plants resistant to RNA viruses such as Potyviruses, Luteoviruses, and Furoviruses, including specific viruses like Wheat streak mosaic virus, Triticum mosaic virus, Soil bourne mosaic virus, and Barley yellow dwarf virus. This resistance is achieved by introducing double-stranded RNA (dsRNA) constructs targeting genes encoding eIF(iso)4E-2 or eIF4G, resulting in inhibition of viral RNA replication in plants.
The problem addressed by the invention is the need for effective viral resistance in plants, specifically targeting RNA viruses that cause significant crop losses. While previous methods and patents have disclosed mutation or silencing of eIF4E to confer viral resistance in plants, it remained unclear whether targeting other translation initiation factors, eIF(iso)4E-2 or eIF4G, could induce viral resistance in susceptible plants. The invention provides RNA interference targeting these factors to confer broad RNA virus resistance.
The invention encompasses methods of producing transgenic wheat plants by introducing chimeric DNA constructs encoding dsRNA for eIF(iso)4E-2 or eIF4G under plant-expressible promoters, producing stable inheritance of the resistance trait across generations. Additionally, non-transgenic wheat plants with mutations silencing eIF(iso)4E-2 or eIF4G alleles are disclosed, produced by chemical, radiation, or targeted genome editing mutagenesis such as EMS, fast neutron, zinc finger nucleases, TALEN, or CRISPR. The plants produced exhibit significant resistance to multiple RNA viruses.
Claims Coverage
The patent contains one independent method claim and one independent composition claim. The following inventive features are identified from these independent claims.
Method of producing virus-resistant plant germplasm by dsRNA targeting eIF(iso)4E-2
A method comprising: (1) introducing into a parental wheat plant germplasm a chimeric DNA molecule containing (i) a plant expressible promoter, (ii) a region encoding dsRNA for eIF(iso)4E-2 that inhibits RNA viral replication, and (iii) a plant translation termination signal; (2) transforming the parental plant germplasm; (3) generating transformed plant germplasm; and (4) selecting germplasm exhibiting immunity to viral RNA replication, wherein resistance is due to inhibited expression of the eIF(iso)4E-2 gene encoding the protein of SEQ. ID. NO: 2.
Transgenic wheat plant resistant to RNA viruses containing dsRNA construct for eIF(iso)4E-2
A transgenic wheat plant genetically transformed to include a chimeric DNA molecule encoding a double stranded RNA molecule targeting eIF(iso)4E-2, conferring resistance to RNA viruses through inhibited expression of the eIF(iso)4E-2 gene encoding the protein of SEQ. ID. NO: 2.
The claims cover methods of producing wheat plant germplasm resistant to RNA viruses by introducing dsRNA constructs targeting the eIF(iso)4E-2 gene, as well as transgenic wheat plants containing these constructs conferring viral resistance through knock-down of eIF(iso)4E-2 expression.
Stated Advantages
The dsRNA constructs targeting eIF(iso)4E-2 or eIF4G confer stable and inherited resistance to multiple RNA viruses, including Potyviruses, Luteoviruses, and Furoviruses, in wheat plants.
Resistance phenotype remains stable across multiple generations of transgenic plants.
The invention allows for production of non-transgenic virus-resistant plants through mutation breeding or targeted genome editing, providing alternative germplasm options without transgenic modifications.
Documented Applications
Producing transgenic wheat plants resistant to RNA viruses such as Wheat streak mosaic virus, Triticum mosaic virus, Soil bourne mosaic virus, and Barley yellow dwarf virus by introducing dsRNA constructs targeting eIF(iso)4E-2 or eIF4G.
Generating non-transgenic wheat plants with mutations in eIF(iso)4E-2 or eIF4G alleles conferring resistance to the same RNA viruses via chemical mutagenesis, radiation, or targeted genome editing.
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