# US 10,306,850 B2 · Yield QTLs in cucumber plants

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- Number: US 10,306,850 B2
- Document ID: US10306850B2
- Status: Granted
- Kind: B2
- Application number: 15/519,446
- Filed: 2015-10-14
- Published: 2017-08-24
- Granted: 2019-06-04
- Earliest claimed: 2014-10-16
- Assignees: Nunhems B.V.
- Applicants: Nunhems B.V.
- Inventors: Gerhard T. M. Reuling; Peter Arnold Gijsbert Kraan; Frank Beenders; Marion Van De Wal; Freddy Hermans; Hans-Peter Koelewijn; Steven D. Tanksley; Alexandra M. Casa
- Web page: https://patentgrep.com/patent/US10306850B2

## CPC

- A01H 6/346: {Cucumis sativus[cucumber]}
- A01H 1/021: {Methods of breeding using interspecific crosses, i.e. interspecies crosses}
- A01H 1/045: {using molecular markers}
- A01H 5/08: Fruits
- 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
- 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 cultivated cucumber plants comprising a yield QTL on chromosome 2 and/or chromosome 6 of their genome.

## Claims

### Claim 1

A Cucumis sativus var. sativus plant, or part thereof, comprising an introgression fragment on chromosome 6 in homozygous or heterozygous form, wherein said introgression fragment is the fragment as found on chromosome 6 in seeds of which a representative sample has been deposited under accession number NCIB 42262, or a smaller fragment derived therefrom, and confers an increase in cucumber fruit yield, and wherein said introgression fragment or smaller fragment comprises at least 1 of the following markers:
a) the AG or AA genotype for the Single Nucleotide Polymorphism marker SNP 12 in SEQ ID NO: 12;
b) the AG or AA genotype for the Single Nucleotide Polymorphism marker SNP 13 in SEQ ID NO: 13;
c) the AG or GG genotype for the Single Nucleotide Polymorphism marker SNP 14 in SEQ ID NO: 14;
d) the CT or TT genotype for the Single Nucleotide Polymorphism marker SNP 15 in SEQ ID NO: 15;
e) the AG or AA genotype for the Single Nucleotide Polymorphism marker SNP 16 in SEQ ID NO: 16;
f) the CT or TT genotype for the Single Nucleotide Polymorphism marker SNP 17 in SEQ ID NO: 17;
g) the CT or CC genotype for the Single Nucleotide Polymorphism marker SNP 18 in SEQ ID NO: 18;
h) the AC or AA genotype for the Single Nucleotide Polymorphism marker SNP 19 in SEQ ID NO: 19;
i) the AC or AA genotype for the Single Nucleotide Polymorphism marker SNP 20 in SEQ ID NO: 20;
j) the AG or GG genotype for the Single Nucleotide Polymorphism marker SNP 21 in SEQ ID NO: 21;
k) the CT or CC genotype for the Single Nucleotide Polymorphism marker SNP 22 in SEQ ID NO: 22;
I) the AG or AA genotype for the Single Nucleotide Polymorphism marker SNP 23 in SEQ ID NO: 23;
m) the CT or CC genotype for the Single Nucleotide Polymorphism marker SNP 24 in SEQ ID NO: 24;
n) the AG or GG genotype for the Single Nucleotide Polymorphism marker SNP 25 in SEQ ID NO: 25;
o) the CT or CC genotype for the Single Nucleotide Polymorphism marker SNP 26 in SEQ ID NO: 26;
p) the AG or AA genotype for the Single Nucleotide Polymorphism marker SNP 27 in SEQ ID NO: 27;
q) the CT or CC genotype for the Single Nucleotide Polymorphism marker SNP 28 in SEQ ID NO: 28;
r) the CT or CC genotype for the Single Nucleotide Polymorphism marker SNP 29 in SEQ ID NO: 29; or
s) the GT or TT genotype for the Single Nucleotide Polymorphism marker SNP 30 in SEQ ID NO: 30.

### Claim 2 (depends on 1)

The plant according to claim 1, wherein said increase in cucumber fruit yield is phenotypically expressed as a significantly higher average number of fruits per plant (FrPP) of the plant line comprising the introgression fragment compared to the genetic control line lacking the introgression fragment when grown under the same environment and/or a significantly higher average fruit weight per plant (GrPP) of the plant line comprising the introgression fragment compared to a genetic control line lacking the introgression fragment when grown under the same environment.

### Claim 3 (depends on 1)

The plant according to claim 1, wherein said introgression fragment on chromosome 6 comprises at least two of the following markers:
a) the AG or AA genotype for the Single Nucleotide Polymorphism marker SNP_12 in SEQ ID NO: 12;
b) the AG or AA genotype for the Single Nucleotide Polymorphism marker SNP_13 in SEQ ID NO: 13;
c) the AG or GG genotype for the Single Nucleotide Polymorphism marker SNP_14 in SEQ ID NO: 14;
d) the CT or TT genotype for the Single Nucleotide Polymorphism marker SNP_15 in SEQ ID NO: 15;
e) the AG or AA genotype for the Single Nucleotide Polymorphism marker SNP_16 in SEQ ID NO: 16;
f) the CT or TT genotype for the Single Nucleotide Polymorphism marker SNP_17 in SEQ ID NO: 17;
g) the CT or CC genotype for the Single Nucleotide Polymorphism marker SNP_18 in SEQ ID NO: 18;
h) the AC or AA genotype for the Single Nucleotide Polymorphism marker SNP_19 in SEQ ID NO: 19;
i) the AC or AA genotype for the Single Nucleotide Polymorphism marker SNP_20 in SEQ ID NO: 20;
j) the AG or GG genotype for the Single Nucleotide Polymorphism marker SNP_21 in SEQ ID NO: 21;
k) the CT or CC genotype for the Single Nucleotide Polymorphism marker SNP_22 in SEQ ID NO: 22;
l) the AG or AA genotype for the Single Nucleotide Polymorphism marker SNP_23 in SEQ ID NO: 23;
m) the CT or CC genotype for the Single Nucleotide Polymorphism marker SNP_24 in SEQ ID NO: 24;
n) the AG or GG genotype for the Single Nucleotide Polymorphism marker SNP_25 in SEQ ID NO: 25;
o) the CT or CC genotype for the Single Nucleotide Polymorphism marker SNP_26 in SEQ ID NO: 26;
p) the AG or AA genotype for the Single Nucleotide Polymorphism marker SNP_27 in SEQ ID NO: 27;
q) the CT or CC genotype for the Single Nucleotide Polymorphism marker SNP_28 in SEQ ID NO: 28;
r) the CT or CC genotype for the Single Nucleotide Polymorphism marker SNP_29 in SEQ ID NO: 29; and/or
s) the GT or TT genotype for the Single Nucleotide Polymorphism marker SNP_30 in SEQ ID NO: 30.

### Claim 4 (depends on 1)

The plant according to claim 1, wherein said introgression fragment on chromosome 6 comprises at least 2 consecutive markers from the following markers:
a) the AG or AA genotype for the Single Nucleotide Polymorphism marker SNP_12 in SEQ ID NO: 12;
b) the AG or AA genotype for the Single Nucleotide Polymorphism marker SNP_13 in SEQ ID NO: 13;
c) the AG or GG genotype for the Single Nucleotide Polymorphism marker SNP_14 in SEQ ID NO: 14;
d) the CT or TT genotype for the Single Nucleotide Polymorphism marker SNP_15 in SEQ ID NO: 15;
e) the AG or AA genotype for the Single Nucleotide Polymorphism marker SNP_16 in SEQ ID NO: 16;
f) the CT or TT genotype for the Single Nucleotide Polymorphism marker SNP_17 in SEQ ID NO: 17;
g) the CT or CC genotype for the Single Nucleotide Polymorphism marker SNP_18 in SEQ ID NO: 18;
h) the AC or AA genotype for the Single Nucleotide Polymorphism marker SNP_19 in SEQ ID NO: 19;
i) the AC or AA genotype for the Single Nucleotide Polymorphism marker SNP_20 in SEQ ID NO: 20;
j) the AG or GG genotype for the Single Nucleotide Polymorphism marker SNP_21 in SEQ ID NO: 21;
k) the CT or CC genotype for the Single Nucleotide Polymorphism marker SNP_22 in SEQ ID NO: 22;
I) the AG or AA genotype for the Single Nucleotide Polymorphism marker SNP_23 in SEQ ID NO: 23;
m) the CT or CC genotype for the Single Nucleotide Polymorphism marker SNP_24 in SEQ ID NO: 24;
n) the AG or GG genotype for the Single Nucleotide Polymorphism marker SNP_25 in SEQ ID NO: 25;
o) the CT or CC genotype for the Single Nucleotide Polymorphism marker SNP_26 in SEQ ID NO: 26;
p) the AG or AA genotype for the Single Nucleotide Polymorphism marker SNP_27 in SEQ ID NO: 27;
q) the CT or CC genotype for the Single Nucleotide Polymorphism marker SNP_28 in SEQ ID NO: 28;
r) the CT or CC genotype for the Single Nucleotide Polymorphism marker SNP_29 in SEQ ID NO: 29; and
s) the GT or TT genotype for the Single Nucleotide Polymorphism marker SNP_30 in SEQ ID NO: 30.

### Claim 5 (depends on 1)

The plant according to claim 1, wherein the introgression fragment on chromosome 6 comprises at least 1 of the following markers:
the AG or AA genotype for the Single Nucleotide Polymorphism marker SNP_12 in SEQ ID NO: 12;
the AG or AA genotype for the Single Nucleotide Polymorphism marker SNP_13 in SEQ ID NO: 13;
the CT or CC genotype for the Single Nucleotide Polymorphism marker SNP_18 in SEQ ID NO: 18;
the AC or AA genotype for the Single Nucleotide Polymorphism marker SNP_19 in SEQ ID NO: 19;
the AC or AA genotype for the Single Nucleotide Polymorphism marker SNP_20 in SEQ ID NO: 20;
the AG or GG genotype for the Single Nucleotide Polymorphism marker SNP_21 in SEQ ID NO: 21;
the CT or CC genotype for the Single Nucleotide Polymorphism marker SNP_22 in SEQ ID NO: 22;
the AG or AA genotype for the Single Nucleotide Polymorphism marker SNP_23 in SEQ ID NO: 23;
the CT or CC genotype for the Single Nucleotide Polymorphism marker SNP_24 in SEQ ID NO: 24;
the AG or GG genotype for the Single Nucleotide Polymorphism marker SNP_25 in SEQ ID NO: 25;
the CT or CC genotype for the Single Nucleotide Polymorphism marker SNP_26 in SEQ ID NO: 26;
the CT or CC genotype for the Single Nucleotide Polymorphism marker SNP_28 in SEQ ID NO: 28; and/or
the GT or TT genotype for the Single Nucleotide Polymorphism marker SNP_30 in SEQ ID NO: 30.

### Claim 6 (depends on 1)

The plant according to claim 1, wherein the introgression fragment on chromosome 6 comprises at least 1 of the following markers:
the AG or AA genotype for the Single Nucleotide Polymorphism marker SNP_13 in SEQ ID NO: 13;
the CT or CC genotype for the Single Nucleotide Polymorphism marker SNP_18 in SEQ ID NO: 18; and/or
the CT or CC genotype for the Single Nucleotide Polymorphism marker SNP_28 in SEQ ID NO: 28.

### Claim 7 (depends on 1)

The plant according to claim 1, wherein the plant is of one of the following cucumber types: pickling cucumbers, slicing cucumbers, long cucumbers, short cucumbers, European greenhouse cucumbers, Beit-Alpha type cucumbers, oriental trellis type cucumbers, or Asian cucumbers.

### Claim 8 (depends on 1)

The plant according to claim 1, wherein the plant is a single cross F1 hybrid or an inbred line.

### Claim 9 (depends on 1)

The plant according to claim 1, wherein the plant is a suitable for once-over mechanical harvest.

### Claim 10 (depends on 1)

The plant according to claim 1, wherein the plant is not a wild cucumber plant or a landrace, or wherein the plant is a cultivated cucumber of the Eurasian cucumber group, the East Asian cucumber group or the Xishuangbanna cucumber group.

### Claim 11 (depends on 1)

The plant according to claim 1, wherein the plant produces seedless fruits without pollination.

### Claim 12 (depends on 1)

The plant according to claim 1, wherein the plant is primarily or entirely gynoecious.

### Claim 13 (depends on 1)

The plant according to claim 1, wherein the plant is uniform regarding the morphological characteristics of the fruits produced by said plant.

### Claim 14 (depends on 1)

The plant according to claim 1, wherein the plant produces fruits which at edible maturity have an average fruit length of at least 10 cm and/or a fruit length to diameter ratio of 2 or more.

### Claim 15 (depends on 1)

The plant according to claim 1, wherein said introgression fragment on chromosome 6 is the fragment as found on chromosome 6 in seeds deposited under accession number NCIMB42262.

### Claim 16 (depends on 1)

The plant according to claim 1, wherein said introgression fragment on chromosome 6 is equal to or less than 10 Mb in size, equal to or less than 8 Mb in size, or equal to or less than 3 Mb in size.

### Claim 17 (depends on 16)

The plant according to claim 16, wherein said introgression fragment is at least 0.5 Mb or at least 1 Mb in size.

### Claim 18 (depends on 1)

The plant according to claim 1, wherein the plant is an indeterminate cucumber.

### Claim 19 (depends on 1)

Seeds from which a plant according to claim 1 can be grown.

### Claim 20 (depends on 1)

A cucumber fruit harvested from a plant according to claim 1.

### Claim 21 (depends on 20)

The fruit according to claim 20, wherein the fruit is bitter free at edible harvest.

### Claim 22 (depends on 20)

The fruit according to claim 20, wherein the fruit has a thin skin at edible harvest.

### Claim 23 (depends on 1)

A plant cell, tissue or plant part of the plant according to claim 1.

### Claim 24

A method for identifying a cultivated C. sativus var. sativus plant comprising an introgression fragment on chromosome 6, wherein said introgression fragment is as found in NCIMB 42262, or a smaller fragment derived therefrom, comprising:
a) screening a population of cultivated C. sativus var. sativus plants using a molecular marker assay which detects at least one SNP marker of: SNP_12 to SNP_30 for detecting the introgression fragment on chromosome 6; and
b) identifying and/or selecting a plant comprising:
i) at least 1 of the SNP markers of SNP_12 to SNP_30 for detecting the introgression fragment on chromosome 6; or
ii) at least 2 consecutive markers from SNP_12 to SNP_30 for detecting the introgression fragment on chromosome 6; or
iii) at least 1 marker from SNP_12, SNP_13, SNP_18 to SNP_26, SNP_28 and/or SNP_30 for detecting the introgression fragment on chromosome 6; or
iv) at least 1, 2 or 3 markers selected from SNP_13, SNP_18 and SNP_28 for detecting the introgression fragment on chromosome 6.

### Claim 25

A method of producing C. sativus F1 hybrid plants comprising an introgression fragment conferring enhanced fruit yield comprising:
a) crossing a first inbred cucumber plant comprising a recombinant chromosome 6 in homozygous form having an introgression fragment comprising an allele conferring enhanced yield, wherein said introgression fragment is as in NCIMB 42262 with a second inbred cucumber plant, and
b) collecting F1 hybrid seeds from said cross.

### Claim 26

A method for generating progeny of NCIMB 42262, said method comprising:
a) selfing a plant grown from seeds deposited under accession number NCIMB 42262 one or more times or crossing said plant one or more times with another cucumber plant to generate progeny seeds;
b) screening said progeny seeds or plants grown from said progeny seeds or parts of the progeny seeds or plants using a molecular marker assay which detects at least one of: SNP_12 to SNP_30 for detecting the introgression fragment on chromosome 6, and
c) identifying and/or selecting a progeny plant comprising:
i) at least 1 of the SNP markers of SNP_12 to SNP_30 for detecting the introgression fragment on chromosome 6; or
ii) at least 2 consecutive markers selected from SNP_12 to SNP_30 for detecting the introgression fragment on chromosome 6; or
iii) at least 1 marker from SNP_12, SNP_13, SNP_18 to SNP_26, SNP_28 and SNP_30 for detecting the introgression fragment on chromosome 6; or
iv) at least 1 marker selected from SNP_13, SNP_18 and SNP_28 for detecting the introgression fragment on chromosome 6.

### Claim 27 (depends on 26)

A progeny plant generated by the method of claim 26, wherein said progeny plant comprises an introgression fragment on chromosome 6 in homozygous or heterozygous form, wherein said introgression fragment is the fragment as found on chromosome 6 in seeds of which a representative sample has been deposited under accession number NCIMB 42262, or a smaller fragment derived therefrom.
