# US 10,766,935 B2 · Plant traits conferred by isolated polynucleotides and polypeptides

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- Number: US 10,766,935 B2
- Document ID: US10766935B2
- Status: Granted
- Kind: B2
- Application number: 16/066,325
- Filed: 2016-12-22
- Published: 2019-03-21
- Granted: 2020-09-08
- Earliest claimed: 2015-12-28
- Assignees: Evogene Ltd.
- Applicants: Evogene Ltd.
- Inventors: Yaacov Micha Brog; Yael Galon Wolfenson; Shlomo Zev Goren; Dror Hilman; Hagai Karchi; Michal Lieberman-Lazarovich; Ronit Rimon Knopf; Ruth Van-Oss Pinhasi
- Web page: https://patentgrep.com/patent/US10766935B2

## CPC

- C07K 14/415: from plants
- C12N 15/8261: {with agronomic (input) traits, e.g. crop yield}
- C12N 15/827: {Flower development or morphology, e.g. flowering promoting factor [FPF]}
- C12N 15/8273: {for drought, cold, salt resistance}
- Y02A 40/146: Genetically Modified [GMO] plants, e.g. transgenic plants
- 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)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE

## Abstract

Provided are isolated polypeptides, isolated polynucleotides encoding same, nucleic acid constructs comprising same, transgenic cells expressing same, transgenic plants expressing same and method of using same for increasing yield, abiotic stress tolerance, growth rate, biomass, vigor, oil content, photosynthetic capacity, seed yield, fiber yield, fiber quality, fiber length, early flowering, grain filling period, harvest index, plant height, and/or nitrogen use efficiency of a plant.

## Claims

### Claim 1

A method of increasing at least one trait selected from the group consisting of biomass, plot coverage, rosette area, rosette diameter photosynthetic capacity, nitrogen use efficiency, and tolerance to nitrogen deficiency stress of a plant as compared to a control plant, comprising over-expressing within the plant a polypeptide at least 95% identical to an amino acid sequence set forth by SEQ ID NO: 15889 as compared to a control plant of the same species which is grown under the same growth conditions, wherein said amino acid sequence comprises the domains depicted by InterPro numbers IPR032675 and IPR001810, thereby increasing the at least one trait selected from biomass, plot coverage, rosette area, rosette diameter, photosynthetic capacity, nitrogen use efficiency, and tolerance to nitrogen deficiency of the plant as compared to the control plant.

### Claim 2

A method of increasing at least one trait selected from the group consisting of biomass, plot coverage, rosette area, rosette diameter photosynthetic capacity, nitrogen use efficiency, and tolerance to nitrogen deficiency stress of a plant as compared to a control plant, comprising over-expressing within the plant a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 15889, 17881 and 17882 as compared to a control plant of the same species which is grown under the same growth conditions, thereby increasing the at least one trait selected from biomass, plot coverage, rosette area, rosette diameter, photosynthetic capacity, nitrogen use efficiency, and tolerance to nitrogen deficiency of the plant as compared to the control plant.

### Claim 3

A method of producing a crop, comprising growing a crop plant over-expressing a polypeptide comprising an amino acid sequence at least 95% identical to the amino acid sequence set forth by SEQ ID NO: 15889 as compared to a control plant of the same species which is grown under the same growth conditions, wherein said amino acid sequence comprises the domains depicted by InterPro numbers IPR032675 and IPR001810, wherein the crop plant is derived from plants which have been subjected to genome editing for over-expressing said polypeptide and/or which have been transformed with an exogenous polynucleotide encoding said polypeptide and which have been selected for at least one trait selected from the group consisting of increased biomass, increased plot coverage, increased rosette area, increased rosette diameter, increased photosynthetic capacity, increased nitrogen use efficiency, and/or increased tolerance to nitrogen deficiency stress as compared to a control plant of the same species which is grown under the same growth conditions, and the crop plant has the at least one trait selected from increased biomass, increased plot coverage, increased rosette area, increased rosette diameter, increased photosynthetic capacity, increased nitrogen use efficiency, and/or increased tolerance to nitrogen deficiency-stress, thereby producing the crop.

### Claim 4 (depends on 2)

The method of claim 2, wherein said polypeptide is expressed from an exogenous polynucleotide comprising the nucleic acid sequence selected from the group consisting of SEQ ID NOs: 407, 105, 2975 and 2976.

### Claim 5

A nucleic acid construct comprising an isolated polynucleotide comprising a nucleic acid sequence encoding a polypeptide which comprises an amino acid sequence at least 95% identical to the amino acid sequence set forth by SEQ ID NO: 15889, and a heterologous promoter for directing transcription of said nucleic acid sequence in a host cell, wherein said amino acid sequence comprises the domains depicted by InterPro numbers IPR032675 and IPR001810, and wherein said amino acid sequence is capable of increasing at least one trait selected from the group consisting of biomass, plot coverage, rosette area, rosette diameter, photosynthetic capacity, nitrogen use efficiency, and/or tolerance to nitrogen deficiency stress of a plant.

### Claim 6

A nucleic acid construct comprising an isolated polynucleotide comprising a nucleic acid sequence encoding a polypeptide which comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 15889, 17881 and 17882, and a heterologous promoter for directing transcription of said nucleic acid sequence in a host cell.

### Claim 7 (depends on 6)

The nucleic acid construct of claim 6, wherein said nucleic acid sequence is selected from the group consisting of SEQ ID NOs: 407, 105, and 2975-2976.

### Claim 8 (depends on 5)

A plant cell transformed with the nucleic acid construct of claim 5.

### Claim 9 (depends on 8)

The plant cell of claim 8, wherein said plant cell forms part of a plant.

### Claim 10 (depends on 5)

A transgenic plant comprising the nucleic acid construct of claim 5.

### Claim 11 (depends on 1)

The method of claim 1, further comprising growing the plant over-expressing said polypeptide under nitrogen-limiting conditions.

### Claim 12 (depends on 5)

A method of growing a crop, the method comprising seeding seeds and/or planting plantlets of a plant transformed with the nucleic acid construct of claim 5, wherein the plant is derived from plants which over-express said polypeptide and which have been selected for at least one trait selected from the group consisting of: increased nitrogen use efficiency, increased tolerance to nitrogen deficiency stress, increased biomass, increased plot coverage, increased rosette area, increased rosette diameter, and increased photosynthetic capacity, as compared to a control plant, thereby growing the crop.

### Claim 13 (depends on 1)

The method of claim 1, further comprising selecting a plant having at least one trait selected from the group consisting of increased biomass, plot coverage, rosette area, rosette diameter, photosynthetic capacity, nitrogen use efficiency, and/or tolerance to nitrogen deficiency stress as compared to a control wild type plant of the same species which is grown under the same growth conditions.

### Claim 14

A method of selecting a plant having at least one trait selected from the group consisting of increased biomass, plot coverage, rosette area, rosette diameter, photosynthetic capacity, nitrogen use efficiency, and/or tolerance to nitrogen deficiency stress as compared to a wild type plant of the same species which is grown under the same growth conditions, the method comprising:
(a) providing plants over-expressing a polypeptide comprising an amino acid sequence at least 95% identical to the amino acid sequence set forth by -SEQ ID NO: 15889, wherein said amino acid sequence comprises the domains depicted by InterPro numbers IPR032675 and IPR001810,
(b) selecting from said plants of step (a) a plant having at least one trait selected from increased biomass, plot coverage, rosette area, rosette diameter, photosynthetic capacity, nitrogen use efficiency, and/or tolerance to nitrogen deficiency stress as compared to a wild type plant of the same species which is grown under the same growth conditions,
thereby selecting the plant having the at least one trait selected from increased biomass, plot coverage, rosette area, rosette diameter, photosynthetic capacity, nitrogen use efficiency, and/or tolerance to nitrogen deficiency stress as compared to the wild type plant of the same species which is grown under the same growth conditions.

### Claim 15 (depends on 13)

The method of claim 13, wherein said selecting is performed under non-stress conditions.

### Claim 16 (depends on 13)

The method of claim 13, wherein said selecting is performed under nitrogen deficiency stress conditions.

### Claim 17 (depends on 1)

The method of claim 1, wherein said polypeptide is set forth by SEQ ID NO: 15889.

### Claim 18 (depends on 3)

The method of claim 3, wherein said polypeptide is set forth by SEQ ID NO: 15889.

### Claim 19 (depends on 5)

The nucleic acid construct of claim 5, wherein said polypeptide is set forth by SEQ ID NO: 15889.

### Claim 20 (depends on 6)

A plant cell transformed with the nucleic acid construct of claim 6.

### Claim 21 (depends on 8)

The plant cell of claim 8, wherein said polypeptide is set forth by SEQ ID NO: 15889.

### Claim 22 (depends on 10)

The transgenic plant of claim 10, wherein said polypeptide is set forth by SEQ ID NO: 15889.

### Claim 23 (depends on 12)

The method of claim 12, wherein said polypeptide is set forth by SEQ ID NO: 15889.

### Claim 24 (depends on 14)

The method of claim 14, wherein said polypeptide is set forth by SEQ ID NO: 15889.