# US 11,441,157 B2 · Vitis vinifera with reduced MLO6 and MLO11 expression and increased resistance to powdery mildew

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- Number: US 11,441,157 B2
- Document ID: US11441157B2
- Status: Granted
- Kind: B2
- Application number: 16/887,412
- Filed: 2020-05-29
- Published: 2020-09-17
- Granted: 2022-09-13
- Earliest claimed: 2015-07-08
- Assignees: Fondazione Edmund Mach
- Applicants: Fondazione Edmund Mach
- Inventors: Mickael Malnoy; Stefano Pessina; Riccardo Velasco; Michele Perazzolli; Luisa Lenzi
- Web page: https://patentgrep.com/patent/US11441157B2

## CPC

- C12N 15/8282: {for fungal resistance}
- A01H 1/1255: {for fungal resistance}
- A01H 6/88: Vitaceae, e.g. Vitus [grape]
- C07K 14/415: from plants
- C12N 15/8218: {Antisense, co-suppression, viral induced gene silencing [VIGS], post-transcriptional induced gene silencing [PTGS]}
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- A01H: NEW PLANTS OR {NON-TRANSGENIC} PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)

## Abstract

Provided herein are Vitis vinifera exhibiting Erysiphe necator resistance. In particular, provided herein are Vitis vinifera having in their genome mildew resistance locus O (MLO) genes, in particular and MLO6 gene and an MLO11 gene, where the MLO6 gene and MLO11 gene have reduced expression and/or function.

## Claims

### Claim 1

An isolated Vitis vinifera comprising in its genome:
a first modification to a VvMLO6 gene encoding a protein having an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 3, the modification to the VvMLO6 gene comprising one or more non-natural mutations, insertions, substitutions, or deletions, wherein the modification results in a reduction in expression of the protein having the amino acid sequence having at least 95% sequence identity to SEQ ID NO: 3 compared to a Vitis vinifera lacking said modification in said VvMLO6 gene encoding the protein having an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 3; and
a second modification to a VvMLO11 gene encoding a protein having an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 2, the modification to the VvMLO11 gene comprising one or more non-natural mutations, insertions, substitutions, or deletions, wherein the modification results in a reduction in expression of the protein having the amino acid sequence having at least 95% sequence identity to SEQ ID NO: 2 compared to a Vitis vinifera lacking said modification in said VvMLO11 gene encoding the protein having an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 2,
wherein said Vitis vinifera having said modifications exhibits a reduction in powdery mildew disease severity compared to a Vitis vinifera lacking said modification modifications in said VvMLO6 gene and said VvMLO11 gene.

### Claim 2 (depends on 1)

The isolated Vitis vinifera of claim 1, wherein the Vitis vinifera is a Long-Cluster Brachetto cultivar.

### Claim 3 (depends on 1)

The isolated Vitis vinifera of claim 1, wherein the Vitis vinifera is a transgenic plant.

### Claim 4 (depends on 1)

The isolated Vitis vinifera of claim 1, wherein the Vitis vinifera is a cultivated plant.

### Claim 5 (depends on 1)

A seed, fruit, plant part, or propagation material of the isolated Vitis vinifera of claim 1, wherein the seed, fruit, plant part, or propagation material comprises the modified gene comprising one or more non-natural mutations, insertions, substitutions, or deletions resulting in:
a decrease of function, loss of function, reduced expression, or absence of a protein having at least 95% sequence identity to SEQ ID NO: 3; and
a decrease of function, loss of function, reduced expression, or absence of a protein having at least 95% sequence identity to SED ID NO: 2.

### Claim 6

A method for obtaining a Vitis vinifera having resistance to powdery mildew comprising introducing a modification comprising one or more non-natural mutations, insertions, substitutions, or deletions to a VvMLO6 gene encoding a protein having at least 95% sequence identity to SEQ ID NO: 3 in a Vitis vinifera, wherein the modification results in:
a decrease of function, loss of function, reduced expression, or absence of a protein having at least 95% sequence identity to SEQ ID NO: 3 compared to a Vitis vinifera lacking said modification in said VvMLO6 gene; and
introducing a second modification comprising one or more non-natural mutations, insertions, substitutions, or deletions to a VvMLO11 gene encoding a protein having at least 95% sequence identity to SEQ ID NO: 2 in a Vitis vinifera, wherein the second modification results in:
a decrease of function, loss of function, reduced expression, or absence of a protein having at least 95% sequence identity to SEQ ID NO: 2 compared to a Vitis vinifera lacking said modification in said VvMLO11 gene.

### Claim 7 (depends on 6)

The method of claim 6, wherein the resistance to powdery mildew is resistance to powdery mildew caused by Erysiphe necator.

### Claim 8 (depends on 6)

The method of claim 6, wherein the modified gene results in at least a 10% reduction in expression of the protein having at least 95% sequence identity to SEQ ID NO: 3 compared to a Vitis vinifera lacking said modification in said VvMLO6 gene.