Plant-mediated silencing of a fatty acid and retinoid binding protein in Pratylenchus penetrans
Inventors
KAMO, KATHRYN K. • Vieira, Paulo C. R. • EISENBACK, JONATHAN D.
Assignees
Virginia Tech Intellectual Properties Inc • US Department of Agriculture USDA • Virginia Polytechnic Institute and State University
Publication Number
US-10407681-B2
Publication Date
2019-09-10
Expiration Date
2038-03-27
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Abstract
The present disclosure provides compositions and methods utilizing double strand ribonucleic acid (dsRNA) to control parasitic nematodes, including Pratylenchus penetrans. More particularly, the present invention relates to several specific synthetic dsRNAs that induce RNA interference (RNAi) in the target nematodes and methods of delivering the dsRNAs to them, such as allowing feeding on plants containing the dsRNAs.
Core Innovation
The invention discloses compositions and methods for controlling parasitic nematodes, particularly Pratylenchus penetrans, using double-stranded ribonucleic acid (dsRNA). Specifically, the patent describes synthetic dsRNAs that target fatty acid- and retinoid-binding protein genes unique to nematodes, inducing RNA interference (RNAi) and silencing gene expression critical for nematode parasitism. These particular dsRNAs correspond to sequences with at least 95% identity to portions of SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3.
The problem addressed is the significant agricultural damage caused by root lesion nematodes like Pratylenchus penetrans, which infect a wide variety of economically important crop, fruit, and ornamental plants. Existing RNAi approaches face challenges with effective delivery of dsRNA to target nematodes, as mechanical microinjection and soaking methods are impractical for large-scale or agricultural use.
The invention provides solutions through plant-mediated delivery of dsRNAs—either by expressing the dsRNAs within transgenic plants or by external application of dsRNAs, allowing uptake and distribution throughout the plant. When nematodes feed on these dsRNA-containing plants, ingestion of the dsRNA induces RNAi and results in decreased nematode survival, development, or reproduction. This targeted approach leverages the specificity of nematode FAR genes to minimize undesirable effects on non-target organisms.
Claims Coverage
The claims cover several inventive features, primarily focusing on dsRNA compositions targeting nematode FAR genes, their expression or presence in plants, DNA constructs encoding these dsRNAs, and methods of inducing RNAi in nematodes.
Double-stranded ribonucleic acid (dsRNA) composition targeting nematode FAR genes
A dsRNA comprising a first strand with at least 95% sequence identity to a portion of at least 19 consecutive nucleotides of SEQ ID NO:1 or SEQ ID NO:3, and a complementary second strand. - The first strand may have higher sequence identity (e.g., at least 99%). - Variations include the first strand matching SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3.
dsRNA expressed in or distributed throughout plants
The dsRNA as described may be expressed in a plant cell, or distributed throughout at least part of a living plant, including specific crops such as corn, soybean, or lily.
Induction of RNA interference (RNAi) in nematodes by ingestion
A method of inducing RNAi in a nematode, including Pratylenchus penetrans, by allowing the nematode to feed on a plant comprising the dsRNA so that the nematode ingests the dsRNA, leading to gene silencing effects.
DNA molecule for dsRNA expression in host cells
A DNA molecule comprising a functional promoter for a host cell and DNA encoding the dsRNA as above, with the host cell potentially being a plant cell.
Transgenic plants, plant cells, or seeds comprising the dsRNA or DNA construct
A plant cell, plant, or seed containing the dsRNA sequences or the DNA molecule encoding the dsRNA, enabling stable or transient expression and subsequent nematode control.
The inventive features cover dsRNA compositions for nematode-specific gene silencing, their plant-based delivery methods, the necessary genetic constructs, and practical applications for crop protection against Pratylenchus penetrans.
Stated Advantages
Provides a targeted method to control parasitic nematodes without affecting non-target organisms such as plants, mammals, or beneficial insects.
Allows for practical, plant-mediated delivery of dsRNA for nematode control, which overcomes limitations of microinjection or soaking methods in agricultural settings.
Reduces nematode numbers and development on treated plants, leading to decreased crop damage.
Documented Applications
Application in controlling Pratylenchus penetrans in crops such as corn, soybean, and lily by plant-mediated dsRNA delivery.
Use in generating transgenic plants or roots expressing nematode-specific dsRNAs for enhanced resistance to nematode infection.
Direct application of dsRNA or dsRNA-producing microbes to plants for uptake and subsequent nematode control.
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