# US 8,541,650 B2 · Plants having altered agronomic characteristics under nitrogen limiting conditions and related constructs and methods involving genes encoding LNT1 polypeptides and homologs thereof

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- Number: US 8,541,650 B2
- Document ID: US8541650B2
- Status: Granted
- Kind: B2
- Application number: 12/999,074
- Filed: 2009-07-08
- Published: 2011-04-28
- Granted: 2013-09-24
- Earliest claimed: 2008-07-08
- Assignees: E. I. Du Pont de Nemours and Company; Pioneer Hi-Bred International, Inc.
- Applicants: Milo Aukerman; Stephen M. Allen; Dale Loussaert; Stanley Luck; Hajime Sakai; Scott V. Tingey
- Inventors: Milo Aukerman; Stephen M. Allen; Dale Loussaert; Stanley Luck; Hajime Sakai; Scott V. Tingey
- Web page: https://patentgrep.com/patent/US8541650B2

## CPC

- C12N 15/8271: {for stress resistance, e.g. heavy metal resistance}
- C12N 15/8241: {Phenotypically and genetically modified plants via recombinant DNA technology}
- C12N 15/8261: {with agronomic (input) traits, e.g. crop yield}
- Y02A 40/146: Genetically Modified [GMO] plants, e.g. transgenic plants
- 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

Isolated polynucleotides and polypeptides and recombinant DNA constructs particularly useful for altering agronomic characteristics of plants under nitrogen limiting conditions, compositions (such as plants or seeds) comprising these recombinant DNA constructs, and methods utilizing these recombinant DNA constructs. The recombinant DNA construct comprises a polynucleotide operably linked to a promoter functional in a plant, wherein said polynucleotide encodes an LNT1 polypeptide.

## Claims

### Claim 1

A method of increasing nitrogen stress tolerance in a 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 encodes a polypeptide having an amino acid sequence of at least 90% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NO: 20;
(b) regenerating a transgenic plant from the regenerable plant cell after step (a); and
(c) selecting for a transgenic plant that exhibits increased nitrogen stress tolerance when compared to a control plant not comprising the recombinant DNA construct.

### Claim 2 (depends on 1)

The method of claim 1, further comprising:
(d) obtaining a progeny plant derived from the transgenic plant, wherein said progeny plant comprises in its genome the recombinant DNA construct and exhibits increased nitrogen stress tolerance when compared to a control plant not comprising the recombinant DNA construct.

### Claim 3

A method of evaluating nitrogen stress tolerance in a 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 encodes a polypeptide having an amino acid sequence of at least 90% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NO:;
(b) regenerating a transgenic plant from the regenerable plant cell after step (a), wherein the transgenic plant comprises in its genome the recombinant DNA construct; and
(c) evaluating the transgenic plant for nitrogen stress tolerance compared to a control plant not comprising the recombinant DNA construct.

### Claim 4 (depends on 3)

The method of claim 3, further comprising:
(d) obtaining a progeny plant derived from the transgenic plant, wherein the progeny plant comprises in its genome the recombinant DNA construct; and
(e) evaluating the progeny plant for nitrogen stress tolerance compared to a control plant not comprising the recombinant DNA construct.

### Claim 5

A method of evaluating nitrogen stress tolerance in a 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 encodes a polypeptide having an amino acid sequence of at least 90% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NO:20;
(b) regenerating a transgenic plant from the regenerable plant cell after step (a), wherein the transgenic plant comprises in its genome the recombinant DNA construct;
(c) obtaining a progeny plant derived from the transgenic plant, wherein the progeny plant comprises in its genome the recombinant DNA construct; and
(d) evaluating the progeny plant for nitrogen stress tolerance compared to a control plant not comprising the recombinant DNA construct.

### Claim 6 (depends on 1)

The method of claim 1, wherein the plant is a maize plant or a soybean plant.

### Claim 7 (depends on 2)

The method of claim 2, wherein the plant is a maize plant or a soybean plant.

### Claim 8 (depends on 3)

The method of claim 3, wherein the plant is a maize plant or a soybean plant.

### Claim 9 (depends on 4)

The method of claim 4, wherein the plant is a maize plant or a soybean plant.

### Claim 10 (depends on 5)

The method of claim 5, wherein the plant is a maize plant or a soybean plant.