# US 2013/0160158 A1 · Manipulation of glutamine synthetases (gs) to improve nitrogen use efficiency and grain yield in higher plants

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- Number: US 2013/0160158 A1
- Document ID: US20130160158A1
- Status: Published application
- Kind: A1
- Application number: 13/772,857
- Filed: 2013-02-21
- Published: 2013-06-20
- Earliest claimed: 2008-10-30
- Assignees: Pioneer Hi-Bred International, Inc.
- Inventors: RAJEEV GUPTA; KANWARPAL SINGH DHUGGA
- Web page: https://patentgrep.com/patent/US20130160158A1

## CPC

- A01H 1/12: {Processes for modifying agronomic input traits, e.g. crop yield}
- C12N 9/93: {Ligases (6)}
- C12N 15/63: Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N 15/8261: {with agronomic (input) traits, e.g. crop yield}
- C12N 15/8271: {for stress resistance, e.g. heavy metal resistance}
- Y02A 40/146: Genetically Modified [GMO] plants, e.g. transgenic plants
- 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)
- Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE

## Abstract

The present invention provides polynucleotides and related polypeptides of the protein GS. The invention provides genomic sequence for the GS gene. GS is responsible for controlling nitrogen utilization efficiency in plants. Glutamine synthase sequences are provided for improving grain yield and plant growth. The invention further provides recombinant expression cassettes, host cells and transgenic plants.

## Claims

### Claim 1

An isolated polynucleotide selected from the group consisting of:
a. a polynucleotide having at least 80% sequence identity, as determined by the GAP algorithm under default parameters, to the full length sequence of a polynucleotide selected from the group consisting of SEQ ID NOS: 43, 45, 47, 49, 51 and 53; wherein the polynucleotide encodes a polypeptide that functions as a modifier of GS;
b. a polynucleotide encoding a polypeptide selected from the group consisting of SEQ ID NO: 44, 46, 48, 50, 52 and 54;
c. a polynucleotide selected from the group consisting of SEQ ID NOS: 43, 45, 47, 49, 51 and 53; and
d. A polynucleotide which is complementary to the polynucleotide of (a), (b) or (c).

### Claim 2 (depends on 1)

A recombinant expression cassette, comprising the polynucleotide of claim 1, wherein the polynucleotide is operably linked, in sense orientation, to a promoter.

### Claim 3 (depends on 2)

A host cell comprising the expression cassette of claim 2.

### Claim 4 (depends on 2)

A transgenic plant comprising the recombinant expression cassette of claim 2.

### Claim 5 (depends on 4)

The transgenic plant of claim 4, wherein said plant is a monocot.

### Claim 6 (depends on 4)

The transgenic plant of claim 4, wherein said plant is a dicot.

### Claim 7 (depends on 4)

The transgenic plant of claim 4, wherein said plant is selected from the group consisting of: maize, soybean, sunflower, sorghum, canola, wheat, alfalfa, cotton, rice, barley, millets, peanut, switchgrass, myscanthus, triticale and cocoa.

### Claim 8 (depends on 4)

A transgenic seed from the transgenic plant of claim 4.

### Claim 9 (depends on 1)

A method of modulating the GS in a plant, comprising:
a. introducing into a plant cell a recombinant expression cassette comprising the polynucleotide of claim 1 operably linked to a promoter;
b. culturing the plant cell under plant cell growing conditions; and
c. regenerating a plant form said plant cell; wherein the GS in said plant is modulated.

### Claim 10 (depends on 9)

The method of claim 9, wherein the plant is selected from the group consisting of: maize, soybean, alfalfa, barley, canola, cocoa, cotton, millets, myscanthus, peanut, rice, rye, sorghum, sugar cane, switchgrass, triticale and wheat.

### Claim 11

A method of decreasing the GS enzyme polypeptide activity in a plant cell, comprising:
a. providing a nucleotide sequence comprising at least 15 consecutive nucleotides of the complement of SEQ ID NO: 43, 45, 47, 49, 51 and 53;
b. providing a plant cell comprising an mRNA having the sequence set forth in SEQ ID NO: 43, 45, 47, 49, 51 and 53; and
c. introducing the nucleotide sequence of step (a) into the plant cell, wherein the nucleotide sequence inhibits expression of the mRNA in the plant cell.

### Claim 12 (depends on 9)

The method of claim 9, wherein said plant cell is from a monocot.

### Claim 13 (depends on 12)

The method of claim 12, wherein said monocot is maize, soybean, alfalfa, barley, canola, cocoa, cotton, millets, myscanthus, peanut, rice, rye, sorghum, sugar cane, switchgrass, triticale or wheat

### Claim 14 (depends on 9)

The method of claim 9, wherein said plant cell is from a dicot.

### Claim 15 (depends on 4)

The transgenic plant of claim 4, wherein the GS enzyme activity in said plant is increased.

### Claim 16 (depends on 15)

The transgenic plant of claim 15, wherein the plant has increased seedling vigor.

### Claim 17 (depends on 15)

The transgenic plant of claim 15, wherein the plant has enhanced silk emergence.

### Claim 18 (depends on 15)

The transgenic plant of claim 15, wherein the plant has enhanced nitrogen assimilation in roots.

### Claim 19 (depends on 15)

The transgenic plant of claim 15, wherein the plant has increased seed number per plant.

### Claim 20 (depends on 15)

The transgenic plant of claim 15, wherein the plant has increased seed size and mass.

### Claim 21 (depends on 15)

The transgenic plant of claim 15, wherein the plant has seed with increased embryo size.

### Claim 22 (depends on 15)

The transgenic plant of claim 15, wherein the plant has increased nitrogen assimilation in the leaf.

### Claim 23 (depends on 15)

The transgenic plant of claim 15, wherein the plant has increased ear size.

### Claim 24 (depends on 15)

The transgenic plant of claim 15, wherein the plant has enhanced nitrogen utilization efficiency.

### Claim 25 (depends on 15)

The transgenic plant of claim 15, wherein the plant has increased nitrogen remobilization during senescence.

### Claim 26 (depends on 15)

The transgenic plant of claim 15, wherein the plant has increased nitrogen remobilization during grain development.