# US 12,600,980 B2 · Disease resistant melon plants

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- Number: US 12,600,980 B2
- Document ID: US12600980B2
- Status: Granted
- Kind: B2
- Application number: 18/002,502
- Filed: 2021-06-25
- Published: 2023-07-20
- Granted: 2026-04-14
- Earliest claimed: 2020-06-29
- Assignees: Syngenta Crop Protection AG
- Applicants: Syngenta Crop Protection AG
- Inventors: Adrien Mauny; Marc Oliver; Bruno Foncelle
- Web page: https://patentgrep.com/patent/US12600980B2

## CPC

- A01H 5/08: Fruits
- C12N 15/8282: {for fungal resistance}
- A01H 1/1255: {for fungal resistance}
- A01H 6/344: {Cucumis melo [melon]}
- C12Q 1/6895: for plants, fungi or algae
- C12Q 2600/13: Plant traits
- C12Q 2600/156: Polymorphic or mutational markers
- 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)
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES

## Abstract

The present invention relates to novel melon plants displaying an increased resistance to Fusarium oxysporum f.sp. melonis race 1,2 infection. The present invention also relates to seeds and parts of said plants, for example fruits. The present invention further relates to methods of making and using such seeds and plants. The present invention also relates to novel genetic sequences associated with said increased resistance and to molecular markers associated with said novel genetic sequences.

## Claims

### Claim 1

A cultivated melon plant, which is a cultivated Cucumis melo plant, resistant to Fusarium oxysporum f.sp. melonis race 1,2 (FOM 1,2) infection, comprising in its genome an introgressed sequence from C. melo subsp. agrestis which confers resistance to FOM 1,2, wherein said introgressed sequence is comprised in melon line 19MFR011167, representative seed of which is deposited under NCIMB Accession No. 43448, or a progeny thereof, is located on chromosome 4 and comprises at least one of the following SNP markers:
a) a G genotype in the heterozygous or homozygous state for SNP marker 1 at a position corresponding to position 95 in SEQ ID NO: 1;
b) an A genotype in the heterozygous or homozygous state for SNP marker 2 at a position corresponding to position 113 in SEQ ID NO: 6;
c) an A genotype in the heterozygous or homozygous state for SNP marker 3 at a position corresponding to position 73 in SEQ ID NO: 11;
d) a G genotype in the heterozygous or homozygous state for SNP marker 4 at a position corresponding to position 239 in SEQ ID NO: 16;
e) an A genotype in the heterozygous or homozygous state for SNP marker 5 at a position corresponding to position 163 in SEQ ID NO: 21;
f) an A genotype in the heterozygous or homozygous state for SNP marker 6 at a position corresponding to position 59 in SEQ ID NO: 26;
g) an A genotype in the heterozygous or homozygous state for SNP marker 7 at a position corresponding to position 119 in SEQ ID NO: 31;
h) an A genotype in the heterozygous or homozygous state for SNP marker 8 at a position corresponding to position 69 in SEQ ID NO: 36;
i) an A genotype in the heterozygous or homozygous state for SNP marker 9 at a position corresponding to position 136 in SEQ ID NO: 41; and/or
j) a T genotype in the heterozygous or homozygous state for SNP marker 10 at a position corresponding to position 108 in SEQ ID NO: 46.

### Claim 2 (depends on 1)

The plant according to claim 1, wherein:
a) the G genotype for SNP marker 1 can be identified in a PCR by amplification of a nucleic acid fragment with a pair of oligonucleotide primers: forward primer of SEQ ID NO: 2 and reverse primer of SEQ ID NO: 5, and probe of SEQ ID NO: 3;
b) the A genotype for SNP marker 2 can be identified in a PCR by amplification of a nucleic acid fragment with a pair of oligonucleotide primers: forward primer of SEQ ID NO: 7 and reverse primer of SEQ ID NO: 10, and probe of SEQ ID NO: 8;
c) the A genotype for SNP marker 3 can be identified in a PCR by amplification of a nucleic acid fragment with a pair of oligonucleotide primers: forward primer of SEQ ID NO: 12 and reverse primer of SEQ ID NO: 15, and probe of SEQ ID NO: 13;
d) the G genotype for SNP marker 4 can be identified in a PCR by amplification of a nucleic acid fragment with a pair of oligonucleotide primers: forward primer of SEQ ID NO: 17 and reverse primer of SEQ ID NO: 20, and probe of SEQ ID NO: 18;
e) the A genotype for SNP marker 5 can be identified in a PCR by amplification of a nucleic acid fragment with a pair of oligonucleotide primers: forward primer of SEQ ID NO: 22 and reverse primer of SEQ ID NO: 25, and probe of SEQ ID NO: 23;
f) the A genotype for SNP marker 6 can be identified in a PCR by amplification of a nucleic acid fragment with a pair of oligonucleotide primers: forward primer of SEQ ID NO: 27 and reverse primer of SEQ ID NO: 30, and probe of SEQ ID NO: 28;
g) the A genotype for SNP marker 7 can be identified in a PCR by amplification of a nucleic acid fragment with a pair of oligonucleotide primers: forward primer of SEQ ID NO: 32 and reverse primer of SEQ ID NO: 35, and probe of SEQ ID NO: 33;
h) the A genotype for SNP marker 8 can be identified in a PCR by amplification of a nucleic acid fragment with a pair of oligonucleotide primers: forward primer of SEQ ID NO: 37 and reverse primer of SEQ ID NO: 40, and probe of SEQ ID NO: 38;
i) the A genotype for SNP marker 9 can be identified in a PCR by amplification of a nucleic acid fragment with a pair of oligonucleotide primers: forward primer of SEQ ID NO: 42 and reverse primer of SEQ ID NO: 45, and probe of SEQ ID NO: 43; and/or
j) the T genotype for SNP marker 10 can be identified in a PCR by amplification of a nucleic acid fragment with a pair of oligonucleotide primers: forward primer of SEQ ID NO: 47 and reverse primer of SEQ ID NO: 50, and probe of SEQ ID NO: 48.

### Claim 3 (depends on 1)

The plant according to claim 1, wherein said introgressed sequence comprises at least one of SEQ ID NO: 1, SEQ ID NO: 6, SEQ ID NO: 11, SEQ ID NO: 16, SEQ ID NO: 21, SEQ ID NO: 26, SEQ ID NO: 31, SEQ ID NO: 36, SEQ ID NO: 41 and/or SEQ ID NO: 46, or a sequence that is at least 80% identical to one or more of said sequences.

### Claim 4 (depends on 1)

The plant according to claim 1, wherein said plant is heterozygous for said at least one SNP marker.

### Claim 5 (depends on 1)

The plant according to claim 1, wherein said plant is homozygous for said at least one SNP marker.

### Claim 6 (depends on 1)

The plant of claim 1, wherein said FOM 1,2 resistance-conferring introgressed sequence confer resistance upon FOM 1,2 wilting strains.

### Claim 7 (depends on 1)

The plant of claim 1, wherein said plant is obtained by crossing melon line 19MFR011167, representative seed of which is deposited under NCIMB Accession No. 43448, or a progeny or an ancestor thereof, with a melon plant that does not contain said FOM 1,2 resistance-conferring introgressed sequence.

### Claim 8 (depends on 1)

The plant of claim 1, wherein said plant is an inbred, a dihaploid or a hybrid plant.

### Claim 9

A plant of melon line 19MFR011167, representative seed of which is deposited under NCIMB Accession No. 43448.

### Claim 10 (depends on 1)

A plant part of a plant according to claim 1.

### Claim 11 (depends on 1)

A seed that produces a plant or a plant part according to claim 1.

### Claim 12 (depends on 1)

A method for producing a cultivated melon plant exhibiting resistance to FOM 1,2 comprising the steps of
a) crossing a plant according to claim 1 with a cultivated melon plant lacking said FOM 1,2 resistance-conferring introgressed sequence;
b) selecting a progeny plant comprising said introgressed sequence located on chromosome 4 conferring resistance to FOM 1,2, said selecting step comprising detecting at least one of the following SNP markers:
i) a G genotype in the heterozygous or homozygous state for SNP marker 1 at a position corresponding to position 95 in SEQ ID NO: 1;
ii) an A genotype in the heterozygous or homozygous state for SNP marker 2 at a position corresponding to position 113 in SEQ ID NO: 6;
iii) an A genotype in the heterozygous or homozygous state for SNP marker 3 at a position corresponding to position 73 in SEQ ID NO: 11;
iv) a G genotype in the heterozygous or homozygous state for SNP marker 4 at a position corresponding to position 239 in SEQ ID NO: 16;
v) an A genotype in the heterozygous or homozygous state for SNP marker 5 at a position corresponding to position 163 in SEQ ID NO: 21;
vi) an A genotype in the heterozygous or homozygous state for SNP marker 6 at a position corresponding to position 59 in SEQ ID NO: 26;
vii) an A genotype in the heterozygous or homozygous state for SNP marker 7 at a position corresponding to position 119 in SEQ ID NO: 31;
viii) an A genotype in the heterozygous or homozygous state for SNP marker 8 at a position corresponding to position 69 in SEQ ID NO: 36;
ix) an A genotype in the heterozygous or homozygous state for SNP marker 9 at a position corresponding to position 136 in SEQ ID NO: 41; and/or
x) a T genotype in the heterozygous or homozygous state for SNP marker 10 at a position corresponding to position 108 in SEQ ID NO: 46;
thereby producing a plant with enhanced resistance to FOM 1,2.

### Claim 13 (depends on 12)

The method according to claim 12, wherein the method further comprises:
a) selfing the selected progeny or crossing the selected progeny with another melon plant to produce further progeny.

### Claim 14 (depends on 13)

The method according to claim 13, wherein further progeny are selected and selfed/crossed for 2 to 10 more generations.

### Claim 15 (depends on 12)

The method according to claim 12, wherein the plant of step a) is melon line 19MFR011167, representative seed of which is deposited under NCIMB Accession No. 43448, or a progeny or an ancestor thereof.

### Claim 16 (depends on 1)

A method for producing a F1 melon plant exhibiting resistance to FOM 1,2, the method comprising crossing an inbred melon plant, which is a plant according to claim 1, with a different inbred melon plant to produce F1 hybrid progeny.

### Claim 17 (depends on 11)

A method of producing melon seed, the method comprising growing a melon plant from the seed of claim 11, and allowing the plant to produce further melon seed.

### Claim 18 (depends on 1)

The cultivated melon plant of claim 1, wherein the cultivated Cucumis melo subsp. melo plant.

### Claim 19 (depends on 1)

The method of claim 1, wherein the cultivated melon plant is a cultivated Cucumis melo subsp. melo plant.