# US 9,434,773 B2 · Plants having stress related traits and methods for making the same

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- Number: US 9,434,773 B2
- Document ID: US9434773B2
- Status: Granted
- Kind: B2
- Application number: 13/617,207
- Filed: 2012-09-14
- Published: 2013-03-14
- Granted: 2016-09-06
- Earliest claimed: 2011-09-14
- Assignees: Dow AgroSciences LLC
- Applicants: Avutu S. Reddy; Sun Yuejin; Donald J. Merlo; Weiting W. Ni; Dayakar Pareddy; Zhifang Gao; Gregory A. Bradfisch
- Inventors: Avutu S. Reddy; Sun Yuejin; Donald J. Merlo; Weiting W. Ni; Dayakar Pareddy; Zhifang Gao; Gregory A. Bradfisch
- Web page: https://patentgrep.com/patent/US9434773B2

## CPC

- C07K 14/415: from plants
- C12N 9/93: {Ligases (6)}
- C12N 15/8273: {for drought, cold, salt resistance}
- C12Y 603/02019: Ubiquitin-protein ligase (6.3.2.19), i.e. ubiquitin-conjugating enzyme
- 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)
- C12Y: ENZYMES

## Abstract

This invention relates generally to a plant cell with increased yield, preferably under condition of transient and repetitive abiotic stress as compared to a corresponding non-transformed wild type plant cell by increasing or generating one or more activities of RING proteins in plants.

## Claims

### Claim 1

A transgenic monocotyledous plant comprising in its genome a stably-integrated, recombinant DNA construct comprising a polynucleotide operably linked to at least one regulatory element, wherein said polynucleotide has a nucleic acid sequence encoding SEQ ID NO: 2, and wherein said monocotyledous plant exhibits increased drought tolerance when compared to a control plant not comprising said recombinant DNA construct.

### Claim 2 (depends on 1)

The transgenic monocotyledous plant of claim 1, wherein the polynucleotide comprises SEQ ID NO: 1 or SEQ ID NO: 3.

### Claim 3 (depends on 1)

The transgenic monocotyledous plant of claim 1, wherein the plant is a maize plant, wheat plant or rice plant.

### Claim 4 (depends on 2)

The transgenic monocotyledous plant of claim 2, wherein the plant is a maize plant, wheat ant or rice plant.

### Claim 5 (depends on 1)

The transgenic monocotyledons plant of claim 1, wherein said plant has an increased yield when grown under drought stress as compared to a non-transformed control plant of the seine species.

### Claim 6 (depends on 1)

The transgenic monocotyledous plant of claim 1, wherein the plant further comprises at least one additional agronomic trait, herbicide trait or insecticide trait.

### Claim 7

A method of increasing drought and/or heat tolerance in a monocotyledous plant, comprising: (a) introducing into a regenerable plant cell a recombinant DNA construct comprising a polynucleotide operably linked to at least one regulatory sequence, wherein the polynucleotide has a nucleic acid sequence encoding SEQ ID NO: 2; and (b) regenerating a transgenic plant from the regenerable plant cell after step (a), wherein the transgenic plant comprises stably-integrated in its genome the recombinant DNA construct and the transgenic plant exhibits increased drought tolerance when compared to a control plant not comprising the recombinant DNA construct.

### Claim 8 (depends on 7)

The method according to claim 7, further comprising: (c) obtaining a progeny plant derived from the transgenic plant, wherein said progeny plant comprises stably-integrated in its genome the recombinant DNA construct and exhibits increased drought tolerance when compared to a control plant not comprising the recombinant DNA construct.

### Claim 9 (depends on 7)

The method according to claim 7, wherein the polynucleotide comprises SEQ ID NO: 1 or SEQ ID NO: 3.

### Claim 10 (depends on 7)

The method according to claim 7, wherein the plant further comprises at least one additional agronomic trait, herbicide trait or insecticide trait.

### Claim 11

A method of improving drought tolerance in a corn line comprising crossing a corn plant of said corn line with a drought tolerant transgenic corn plant having a recombinant DNA expressing a protein having the amino acid sequence of SEQ ID NO:2 to produce a corn seed having the recombinant DNA.

### Claim 12

A method for increasing yield in a corn crop subject to water deficit during its growth, said method comprising planting transgenic seeds having a stably-integrated, recombinant DNA, wherein the stably-integrated, recombinant DNA comprises the nucleic amino acid sequence of SEQ ID NO:1 or SEQ ID NO: 3 or wherein the stably-integrated, recombinant DNA encodes a protein having the amino acid sequence of SEQ ID NO:2 and allowing said seeds to grow to mature corn plants while being subjected to drought during at least one ten day period during vegetative phase of growth.

### Claim 13

A drought tolerant transgenic corn plant having a recombinant DNA expressing a protein having the amino acid sequence of SEQ ID NO:2.

### Claim 14 (depends on 13)

The transgenic corn plant of claim 13, wherein said corn plant further comprises at least one additional agronomic trait, herbicide trait or insecticide trait.

### Claim 15

Transgenic seed for growing a drought tolerant corn plant, said seed comprising in its genome a stably-integrated, recombinant DNA encoding a protein having the amino acid sequence of SEQ ID NO: 2.

### Claim 16

A DNA construct comprising a nucleic acid encoding the polypeptide of SEQ ID NO: 2, wherein the nucleic acid is operably linked to a heterologous regulatory element and wherein SEQ ID NO: 2 increases the maintenance of membrane integrity under abiotic stress in a plant cell.

### Claim 17 (depends on 16)

The DNA construct of claim 16, wherein the nucleic acid has been optimized for expression in a plant.

### Claim 18 (depends on 16)

The DNA construct of claim 16, wherein the nucleic acid comprises the nucleotide sequence of SEQ ID NO: 3.

### Claim 19 (depends on 16)

A plant expression vector comprising the DNA construct of claim 16.

### Claim 20 (depends on 16)

A transgenic plant cell comprising, stably-integrated in its genome, the DNA construct of claim 16.

### Claim 21 (depends on 20)

A transgenic plant cell culture, plant tissue, or whole plant comprising the transgenic plant cell of claim 20.

### Claim 22

A method for making a transgenic monocotyledous plant having an drought related trait relative to a control plant, comprising introducing a recombinant construct into a plant cell, plant, or part thereof, wherein the drought related trait is one or more of: (i) increased early vigor; (ii) increased biomass; (iii) increased total seed yield per plant; (iv) increased seed filling rate; (v) increased harvest index; (vi) increased thousand kernel weight, (vii) increased abiotic stress resistance, or (viii) increased nutrient uptake efficiency, and wherein the construct comprises (a) a nucleic acid sequence encoding the polypeptide of SEQ ID NO: 2; (b) one or more control sequences capable of driving expression of the nucleic acid sequence of (a) and optionally (c) a transcription termination sequence.