# US 12,408,674 B2 · Methods and compositions for controlling plant viral infection

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- Number: US 12,408,674 B2
- Document ID: US12408674B2
- Status: Granted
- Kind: B2
- Application number: 16/977,038
- Filed: 2019-03-08
- Published: 2020-12-31
- Granted: 2025-09-09
- Earliest claimed: 2018-03-09
- Assignees: Monsanto Technology LLC
- Applicants: Monsanto Technology LLC
- Inventors: Charles Hagen
- Web page: https://patentgrep.com/patent/US12408674B2

## CPC

- A01N 63/60: Isolated nucleic acids
- A01H 3/00: Processes for modifying phenotypes {, e.g. symbiosis with bacteria} (A01H4/00 takes precedence)
- A01H 3/04: by treatment with chemicals
- A01N 25/30: characterised by the surfactants
- A01N 57/16: containing heterocyclic radicals
- C12N 15/113: Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; {Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing (when used in plants C12N15/8218)}
- C12N 15/8218: {Antisense, co-suppression, viral induced gene silencing [VIGS], post-transcriptional induced gene silencing [PTGS]}
- C12N 15/8283: {for virus resistance}
- C12N 2770/26021: Viruses as such, e.g. new isolates, mutants or their genomic sequences
- A01N: PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF (preservation of food or foodstuff A23); BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES (preparations for medical, dental or toiletry purposes which kill or prevent the growth or proliferation of unwanted organisms A61K); PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01H: NEW PLANTS OR {NON-TRANSGENIC} PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

Methods and compositions for treatment and prevention of Potexvirus disease in plants are provided, including methods and compositions employing double-stranded RNA polynucleotides and a transfer agent. Further provided are compositions for treatment or prevention of Potexvirus disease in plants, and methods for reducing expression of a Potexvirus gene and for identifying polynucleotides useful in modulating gene expression in plant viruses.

## Claims

### Claim 1

A method of treatment or prevention of a Pepino Mosaic Virus (PepMV) infection in a plant, the method comprising topically applying to said plant a composition comprising a double-stranded RNA polynucleotide and a transfer agent, wherein the double-stranded RNA polynucleotide consists of a first polynucleotide sequence that consists of at least 21 contiguous nucleotides of SEQ ID NO: 14 and a second polynucleotide sequence that is completely complementary to the first polynucleotide sequence, and wherein the symptoms of viral infection or development of symptoms are reduced or eliminated in said plant relative to a plant not treated with said composition when grown under the same conditions.

### Claim 2 (depends on 1)

The method of claim 1, wherein said transfer agent is an organosilicone surfactant composition or compound contained therein.

### Claim 3 (depends on 2)

The method of claim 2, wherein said double-stranded RNA polynucleotide consists of the nucleotide sequence of SEQ ID NO:14.

### Claim 4 (depends on 1)

The method of claim 1, wherein said composition is topically applied by spraying, dusting, or is applied to the plant surface as matrix-encapsulated RNA.

### Claim 5

A composition comprising a double-stranded RNA polynucleotide and a transfer agent, wherein said double-stranded RNA polynucleotide consists of a first polynucleotide sequence that consists of at least 21 contiguous nucleotides of SEQ ID NO: 14 and a second polynucleotide sequence that is completely complementary to the first polynucleotide sequence, and wherein topical application of the composition to a plant reduces or eliminates the symptoms of PepMV infection or development in said plant relative to a plant not treated with said composition when grown under the same conditions.

### Claim 6 (depends on 5)

The composition of claim 5, wherein said transfer agent is an organosilicone composition.

### Claim 7 (depends on 5)

The composition of claim 5, wherein said double-stranded RNA polynucleotide consists of the nucleotide sequence of SEQ ID NO:14.

### Claim 8 (depends on 5)

A method of reducing expression of an essential PepMV gene comprising contacting a PepMV particle with the composition of claim 5.

### Claim 9 (depends on 8)

The method of claim 8, wherein said transfer agent is an organosilicone compound.

### Claim 10 (depends on 8)

The method of claim 8, wherein said double-stranded RNA polynucleotide consists of the nucleotide sequence of SEQ ID NO:14.

### Claim 11 (depends on 5)

An agricultural chemical composition comprising an admixture of the composition of claim 5 and a pesticide.

### Claim 12 (depends on 11)

The agricultural chemical composition of claim 11, wherein said pesticide is selected from the group consisting of anti-viral compounds, insecticides, fungicides, nematocides, bactericides, acaricides, growth regulators, chemosterilants, semiochemicals, repellents, attractants, pheromones, feeding stimulants, and biopesticides.