# US 10,865,422 B2 · Plants with enhanced photosynthesis and methods of manufacture thereof

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- Number: US 10,865,422 B2
- Document ID: US10865422B2
- Status: Granted
- Kind: B2
- Application number: 16/421,599
- Filed: 2019-05-24
- Published: 2019-12-05
- Granted: 2020-12-15
- Earliest claimed: 2013-12-31
- Assignees: University of Massachusetts
- Applicants: The University of Massachusetts
- Inventors: Danny J. Schnell; Mine O. Canakci; Bibin Paulose; Michelle DaCosta Inguagiato
- Web page: https://patentgrep.com/patent/US10865422B2

## CPC

- C12N 15/8269: {Photosynthesis}
- C07K 14/405: from algae
- C12N 15/8245: {involving modified carbohydrate or sugar alcohol metabolism, e.g. starch biosynthesis}
- C12N 15/8247: {involving modified lipid metabolism, e.g. seed oil composition}
- C12N 15/8261: {with agronomic (input) traits, e.g. crop yield}
- C12N 15/8271: {for stress resistance, e.g. heavy metal resistance}
- C12N 15/8273: {for drought, cold, salt resistance}
- Y02A 40/146: Genetically Modified [GMO] plants, e.g. transgenic plants
- Y02E 50/10: Biofuels, e.g. bio-diesel
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- 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)
- Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02E: REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION

## Abstract

A transgenic plant having enhanced photosynthesis is disclosed. The transgenic plant is transformed with a transgenic polynucleotide encoding a heterologous bicarbonate transporter. The bicarbonate transporter can be from an algae or a cyanobacterial species. The transgenic polynucleotide comprises a nucleic acid sequence encoding the bicarbonate transporter under the control of a functional plant promoter and optionally includes a chloroplast envelope targeting peptide heterologous to the bicarbonate transporter. Methods of making the transgenic plant and transgenic polynucleotide are disclosed.

## Claims

### Claim 1

A transgenic plant transformed with a recombinant DNA construct comprising a plant-expressible transcription regulatory sequence operatively linked to a polynucleotide encoding an algal CCP1 or CCP2 polypeptide comprising an amino acid sequence that is at least 75% identical to SEQ ID NO: 6,
wherein the transgenic plant has:
(i) a CO2 assimilation rate higher than a plant of the same species not transformed with the recombinant DNA construct; and
(ii) a reduced transpiration rate lower than a plant of the same species not transformed with the recombinant DNA construct.

### Claim 2 (depends on 1)

The transgenic plant of claim 1, wherein the algae is a Chlamydomonas species.

### Claim 3 (depends on 1)

The transgenic plant of claim 1, wherein the amino acid sequence is at least 85% identical to SEQ ID NO: 6.

### Claim 4 (depends on 1)

The transgenic plant of claim 1, wherein the amino acid sequence is at least 90% identical to SEQ ID NO: 6.

### Claim 5 (depends on 1)

The transgenic plant of claim 1, wherein the amino acid sequence is at least 95% identical to SEQ ID NO: 6.

### Claim 6 (depends on 1)

The transgenic plant of claim 1, wherein the amino acid sequence comprises SEQ ID NO: 6.

### Claim 7 (depends on 1)

The transgenic plant of claim 1, wherein the transgenic plant comprises one or more of Borago officinalis, Brassica campestris, Brassica napus, Brassica rapa, Camelina species, Cannabis sativa, Carthamus tinctorius, Cocos nucifera, Crambe abyssinica, Cuphea species, Elaeis guinensis, Elaeis oleifera, Glycine max, Gossypium hirsutum, Gossypium barbadense, Gossypium herbaceum, Helianthus annuus, Linum usitatissimum, Oenothera biennis, Olea europaea, Oryza sativa, Ricinus communis, Sesamum indicum, Triticum species, Zea mays, walnut or almond.

### Claim 8 (depends on 7)

The transgenic plant of claim 7, wherein the transgenic plant is Camelina sativa.

### Claim 9 (depends on 1)

The transgenic plant of claim 1, wherein:
(i) the CO2 assimilation rate of the transgenic plant is at least 5% higher than a plant of the same species not transformed with the recombinant DNA construct; and
(ii) the reduced transpiration rate of the transgenic plant is at least 5% lower than a plant of the same species not transformed with the recombinant DNA construct.

### Claim 10

A method of producing a transgenic plant having enhanced photosynthesis, the method comprising:
transforming a plant cell with a recombinant polynucleotide comprising a nucleic acid sequence encoding an algal CCP1 or CCP2 polypeptide, comprising an amino acid sequence that is at least 75% identical to SEQ ID NO: 6, operatively linked to a plant-expressible promoter, wherein the nucleic acid sequence encoding the algal CCP1 or CCP2 polypeptide is further operatively linked to a transcription terminator;
growing a transgenic plant from the plant cell until the transgenic plant produces seed; and
selecting seeds from the transgenic plant in which the transgenic plant has:
(i) a CO2 assimilation rate higher than a plant of the same species not transformed with the recombinant DNA construct; and
(ii) a reduced transpiration rate lower than a plant of the same species not transformed with the recombinant DNA construct.

### Claim 11 (depends on 10)

The method of claim 10, wherein the algae is a Chlamydomonas species.

### Claim 12 (depends on 10)

The method of claim 10, wherein the amino acid sequence is at least 85% identical to SEQ ID NO: 6.

### Claim 13 (depends on 10)

The method of claim 10, wherein the amino acid sequence is at least 90% identical to SEQ ID NO: 6.

### Claim 14 (depends on 10)

The method of claim 10, wherein the amino acid sequence is at least 95% identical to SEQ ID NO: 6.

### Claim 15 (depends on 10)

The method of claim 10, wherein the amino acid sequence comprises SEQ ID NO: 6.

### Claim 16 (depends on 10)

The method of claim 10, wherein the transgenic plant comprises one or more of Borago officinalis, Brassica campestris, Brassica napus, Brassica rapa, Camelina species, Cannabis sativa, Carthamus tinctorius, Cocos nucifera, Crambe abyssinica, Cuphea species, Elaeis guinensis, Elaeis oleifera, Glycine max, Gossypium hirsutum, Gossypium barbadense, Gossypium herbaceum, Helianthus annuus, Linum usitatissimum, Oenothera biennis, Olea europaea, Oryza sativa, Ricinus communis, Sesamum indicum, Triticum species, Zea mays, walnut or almond.

### Claim 17 (depends on 16)

The method of claim 16, wherein the transgenic plant is Camelina sativa.

### Claim 18 (depends on 10)

The method of claim 10, wherein:
(i) the CO2 assimilation rate of the transgenic plant is at least 5% higher than a plant of the same species not transformed with the recombinant DNA construct; and
(ii) the reduced transpiration rate of the transgenic plant is at least 5% lower than a plant of the same species not transformed with the recombinant DNA construct.