# US 7,745,696 B2 · Suppression of Tla1 gene expression for improved solar conversion efficiency and photosynthetic productivity in plants and algae

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- Number: US 7,745,696 B2
- Document ID: US7745696B2
- Status: Granted
- Kind: B2
- Application number: 11/423,620
- Filed: 2006-06-12
- Published: 2008-05-22
- Granted: 2010-06-29
- Earliest claimed: 2006-06-12
- Assignees: The Regents of the University of California
- Inventors: Anastasios Melis; Mautusi Mitra
- Web page: https://patentgrep.com/patent/US7745696B2

## CPC

- C12N 15/113: Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; {Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing (when used in plants C12N15/8218)}
- C07K 14/415: from plants
- C12N 15/825: {involving pigment biosynthesis}
- C12N 15/8269: {Photosynthesis}
- C12N 15/8271: {for stress resistance, e.g. heavy metal resistance}
- 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)

## Abstract

The invention provides method and compositions to minimize the chlorophyll antenna size of photosynthesis by decreasing TLA1 gene expression, thereby improving solar conversion efficiencies and photosynthetic productivity in plants, e.g., green microalgae, under bright sunlight conditions.

## Claims

### Claim 1

A method of decreasing chlorophyll antenna size in a strain of green algae, the method comprising:
inhibiting expression of a Tla1 nucleic acid in the green algae strain by introducing into the green algae strain an expression cassette comprising a promoter operably linked to a polynucleotide, or a complement thereof, wherein the polynucleotide is at least 90% identical to at least 200 contiguous nucleotides of a nucleic acid sequence encoding SEQ ID NO:2; or comprises at least 20 contiguous nucleotides of SEQ ID NO:1 or SEQ ID NO:3; and
selecting algae with decreased chlorophyll antenna size compared to green algae in which the expression cassette has not been introduced.

### Claim 2 (depends on 1)

The method of claim 1, wherein the promoter is constitutive.

### Claim 3 (depends on 1)

The method of claim 1, wherein the promoter is inducible.

### Claim 4 (depends on 1)

The method of claim 1, wherein the polynucleotide is operably linked to the promoter in the antisense orientation.

### Claim 5 (depends on 1)

The method of claim 1, wherein the polynucleotide is operably linked to the promoter in the sense orientation.

### Claim 6 (depends on 1)

The method of claim 1, wherein the polynucleotide is an siRNA.

### Claim 7 (depends on 1)

The method of claim 1, wherein the polynucleotide comprises at least 200 contiguous nucleotides of a nucleic acid that encodes SEQ ID NO:2.

### Claim 8 (depends on 1)

The method of claim 1, wherein the nucleic acid that encodes SEQ ID NO:2 has the sequence set forth in SEQ ID NO:3 or SEQ ID NO:1.

### Claim 9 (depends on 1)

The method of claim 1, wherein the algae are selected from Chlamydomonas reinhardtii, Scenedesmus obliquus, Chlorella vulgaris, Botryococcus braunii, Botryococcus sudeticus, Dunaliella salina, or Haematococcus pluvialis.

### Claim 10

A strain of green algae comprising an expression cassette comprising a polynucleotide, or a complement thereof, that is at least 90% percent identical to at least 200 contiguous nucleotides of a sequence encoding SEQ ID NO:2; or comprises at least 20 contiguous nucleotides to SEQ ID NO:1 or SEQ ID NO:3, or the complement thereof.

### Claim 11 (depends on 10)

The strain of green algae of claim 10, wherein the algae are selected from Chlamydomonas reinhardtii, Scenedesmus obliquus, Chlorella vulgaris, Botryococcus braunii, Botryococcus sudeticus, Dunaliella salina, or Haematococcus pluvialis.

### Claim 12 (depends on 10)

A method of enhancing yields of photosynthetic productivity under high-density growth conditions, the method comprising cultivating a plant of claim 10 under bright sunlight and high density growth conditions.

### Claim 13 (depends on 1)

The method of claim 1, wherein the polynucleotide is at least 90% identical to at least 200 contiguous nucleotides of a nucleic acid sequence encoding SEQ ID NO:2.

### Claim 14 (depends on 1)

The method of claim 1, wherein the polynucletoide comprises at least 20 contiguous nucleotides of SEQ ID NO:1or SEQ ID NO:3, or the complement thereof.

### Claim 15 (depends on 10)

The strain of green algae of claim 10, wherein the promoter is constitutive.

### Claim 16 (depends on 10)

The strain of green algae of claim 10, wherein the promoter is inducible.

### Claim 17 (depends on 10)

The strain of green algae of claim 10, wherein the polynucleotide is operably linked to the promoter in the antisense orientation.

### Claim 18 (depends on 10)

The strain of green algae of claim 10, wherein the polynucleotide is operably linked to the promoter in the sense orientation.

### Claim 19 (depends on 10)

The strain of green algae of claim 10, wherein the polynucleotide is an siRNA.

### Claim 20 (depends on 10)

The strain of green algae of claim 10, wherein the polynucleotide is at least 90% percent identical to at least 200 contiguous nucleotides of a sequence encoding SEQ ID NO:2.

### Claim 21 (depends on 10)

The strain of green algae of claim 10, wherein the polynucleotide comprises at least 200 contiguous nucleotides of a nucleic acid that encodes SEQ ID NO:2.

### Claim 22 (depends on 20)

The strain of green algae of claim 20, wherein the nucleic acid that encodes SEQ ID NO:2 has the sequence set forth in SEQ ID NO:3 or SEQ ID NO:1.

### Claim 23 (depends on 10)

The strain of green algae of claim 10, wherein the polynucleotide comprises at least 20 contiguous nucleotides of SEQ ID NO:1 or SEQ ID NO:3, or the complement thereof.