Plant germplasm resistant to RNA viruses
Inventors
Fellers, John • Trick, Harold N. • Cruz, Luisa • Rupp, Jessica
Assignees
US Department of Agriculture USDA
Publication Number
US-10633671-B2
Publication Date
2020-04-28
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 including Potyviruses, Luteoviruses, and Furoviruses. More specifically, it provides resistance to viruses such as Wheat streak mosaic virus, Triticum mosaic virus, Soil bourne mosaic virus, and Barley yellow dwarf virus through the use of dsRNA constructs targeting translation initiation factors eIF(iso)4E-2 and eIF4G.
The invention discloses a method of producing plant germplasm by introducing chimeric DNA molecules encoding dsRNA for eIF(iso)4E-2 or eIF4G under control of plant-expressible promoters, transforming parental plants, and selecting progeny with inhibited expression of these factors conferring viral resistance. Both transgenic plants expressing these constructs and non-transgenic plants harboring mutations in the genes encoding these initiation factors are provided.
The problem addressed is the need to induce viral resistance in susceptible plants by targeting essential genes mediating viral replication. Prior art has linked viral resistance to mutations or silencing of eIF4E; however, there remained an unmet need to determine whether targeting eIF(iso)4E-2 or eIF4G via RNA interference or gene editing could confer resistance to diverse RNA viruses affecting important crops such as wheat.
Claims Coverage
The patent contains one independent method claim and two independent plant composition claims that define the core inventive features related to producing RNA virus-resistant wheat plants by targeting eIF4G through RNA interference.
Method for producing RNA virus-resistant wheat germplasm by introducing chimeric DNA encoding dsRNA for eIF4G
The method involves introducing a chimeric DNA molecule comprising a plant expressible promoter, a region encoding dsRNA specific for eIF4G capable of inhibiting RNA viral replication, and a plant translation termination signal into wheat parental germplasm. It further includes transforming the germplasm, generating transformed plants, and selecting plants demonstrating immunity due to inhibited eIF4G expression encoding the protein of SEQ. ID. NO: 4.
Transgenic wheat plant comprising chimeric DNA encoding dsRNA for eIF4G and exhibiting virus resistance
A transgenic wheat plant that contains a chimeric DNA molecule encoding a double stranded RNA molecule specific for eIF4G, conferring resistance to RNA viruses through reduced eIF4G gene expression, specifically the protein comprising SEQ. ID. NO: 4.
Transgenic wheat plant comprising sequence encoding eIF4G dsRNA or its antisense with increased resistance
A transgenic wheat plant incorporating a nucleotide sequence encoding SEQ. ID. NO: 3 or its antisense, which results in increased resistance to positive strand RNA viruses.
The claims collectively cover the method of generating wheat plants resistant to multiple RNA viruses by RNA interference targeting eIF4G, as well as the transgenic plants themselves incorporating the dsRNA constructs or nucleotide sequences encoding eIF4G segments that mediate resistance.
Stated Advantages
The dsRNA constructs provide sequence-specific inhibition of essential eukaryotic translation initiation factors involved in viral replication, effectively conferring virus resistance.
The method enables stable expression of resistance traits across multiple generations, ensuring durable viral immunity in crop plants.
The approach tolerates sequence variations allowing utilization of dsRNA with high but not necessarily perfect sequence identity to target genes, facilitating broad application across viral strains and plant variations.
Documented Applications
Producing transgenic wheat plants resistant to RNA viruses such as Potyviruses, Luteoviruses, and Furoviruses.
Generating wheat plants resistant to specific viruses including Wheat streak mosaic virus, Triticum mosaic virus, Soil bourne mosaic virus, and Barley yellow dwarf virus.
Producing non-transgenic wheat plants with mutations in eIF(iso)4E-2 or eIF4G genes conferring RNA virus resistance via chemical mutagenesis, radiation, or targeted genome editing approaches like zinc finger nucleases, TALEN, or CRISPR.
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