Granted Patent B2

US 8,952,217 · App. 11/516,230 · Granted Feb 10, 2015

Inventors: Piotr Puzio (Berlin, DE); Birgit Wendel (Berlin, DE); Michael Manfred Herold (Berlin, DE); Ralf Looser (Berlin, DE); Astrid Blau (Stahnsdorf, DE); Gunnar Plesch (Potsdam, DE); Beate Kamlage (Berlin, DE); Florian Schauwecker (Berlin, DE)

Assignee: Metanomics GmbH

C12N15/8214C12N15/8221C12N15/8243C12N15/8245C12N15/8247C12N15/825C12N15/8251C12N15/8253C12N15/8254C12P7/64C12P13/06C12P13/10C12P13/12

Abstract

Claims (68)

1. A process for decreasing the amount of verbascose in a plant cell, plant, or part thereof, which comprises

a) introducing into a chloroplast of a plant cell, plant, or a part thereof a nucleic acid comprising

i) the nucleic acid sequence of SEQ ID NO: 71513;

ii) a nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 71514;

iii) a nucleic acid sequence encoding an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 71514; or

iv) a nucleic acid which hybridizes with the complement of the nucleic acid of (i) under high stringent hybridization conditions at about 0.2×SSC at 50° C. to 65° C.;

b) introducing into a plant cell, plant, or a part thereof a nucleic acid operably linked to a polynucleotide encoding a transit peptide, wherein the nucleic acid comprises

i) the nucleic acid sequence of SEQ ID NO: 71513;

ii) a nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 71514;

iii) a nucleic acid sequence encoding an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 71514; or

iv) a nucleic acid which hybridizes with the complement of the nucleic acid of (i) under high stringent hybridization conditions at about 0.2×SSC at 50° C. to 65° C.; or

c) introducing into a plant cell, plant, or a part thereof a nucleic acid operably linked to a polynucleotide encoding a chloroplast localization sequence, wherein the nucleic acid comprises

i) the nucleic acid sequence of SEQ ID NO: 71513;

ii) a nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 71514;

iii) a nucleic acid sequence encoding an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 71514; or

iv) a nucleic acid which hybridizes with the complement of the nucleic acid of (i) under high stringent hybridization conditions of about 0.2×SSC at 50° C. to 65° C.;

wherein expression of the nucleic acid results in a decrease in the amount of verbascose relative to a corresponding control plant cell, plant, or part thereof.

2. The process of claim 1 , wherein the transit peptide is derived from a protein selected from the group consisting of

ribulose bisphosphate carboxylase/oxygenase, 5-enolpyruvyl-shikimate-3-phosphate synthase, acetolactate synthase, chloroplast ribosomal protein CS 17, Cs protein, ferredoxin, plastocyanin, ribulose bisphosphate carboxylase activase, tryptophan synthase, acyl carrier protein, plastid chaperonin-60, cytochrome c 552 , 22-kDA heat shock protein, 33-kDa Oxygen-evolving enhancer protein 1, ATP synthase γ subunit, ATP synthase δ subunit, chlorophyll-a/b-binding proteinII-1, Oxygen-evolving enhancer protein 2, Oxygen-evolving enhancer protein 3, photosystem I: P21, photosystem I: P28, photosystem I: P30, photosystem I: P35, photosystem I: P37, glycerol-3-phosphate acyltransferases, chlorophyll a/b binding protein, CAB2 protein, hydroxymethyl-bilane synthase, pyruvate-orthophosphate dikinase, CAB3 protein, plastid ferritin, ferritin, early light-inducible protein, glutamate-1-semialdehyde aminotransferase, protochlorophyllide reductase, starch-granule-bound amylase synthase, light-harvesting chlorophyll a/b-binding protein of photosystem II, major pollen allergen Lol p 5a, plastid ClpB ATP-dependent protease, superoxide dismutase, ferredoxin NADP oxidoreductase, 28-kDa ribonucleoprotein, 31-kDa ribonucleoprotein, 33-kDa ribonucleoprotein, acetolactate synthase, ATP synthase CF 0 subunit 1, ATP synthase CF 0 subunit 2, ATP synthase CF 0 subunit 3, ATP synthase CF 0 subunit 4, cytochrome f, ADP-glucose pyrophosphorylase, glutamine synthase, glutamine synthase 2, carbonic anhydrase, GapA protein, heat-shock-protein hsp21, phosphate translocator, plastid ClpA ATP-dependent protease, plastid ribosomal protein CL24, plastid ribosomal protein CL9, plastid ribosomal protein PsCL18, plastid ribosomal protein PsCL25, DAHP synthase, starch phosphorylase, root acyl carrier protein II, betaine-aldehyde dehydrogenase, GapB protein, glutamine synthetase 2, phosphoribulokinase, nitrite reductase, ribosomal protein L12, ribosomal protein L13, ribosomal protein L21, ribosomal protein L35, ribosomal protein L40, triose phosphate-3-phosphoglyerate-phosphate translocator, ferredoxin-dependent glutamate synthase, glyceraldehyde-3-phosphate dehydrogenase, NADP-dependent malic enzyme, and NADP-malate dehydrogenase.

3. A process for producing a plant cell, plant or part thereof with a decrease in the amount of verbascose relative to a corresponding control plant cell, plant or part thereof, which comprises

a) introducing into a chloroplast of a plant cell, plant, or a part thereof a nucleic acid comprising

i) the nucleic acid sequence of SEQ ID NO: 71513;

ii) a nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 71514;

iii) a nucleic acid sequence encoding an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 71514; or

iv) a nucleic acid which hybridizes with the complement of the nucleic acid of (i) under high stringent hybridization conditions of about 0.2×SSC at 50° C. to 65° C.;

b) introducing into a plant cell, plant, or a part thereof a nucleic acid comprising a nucleic acid operably linked to a polynucleotide encoding a transit peptide, wherein the nucleic acid comprises

i) the nucleic acid sequence of SEQ ID NO: 71513;

ii) a nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 71514;

iii) a nucleic acid sequence encoding an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 71514; or

iv) a nucleic acid which hybridizes with the complement of the nucleic acid of (i) under high stringent hybridization conditions of about 0.2×SSC at 50° C. to 65° C.; or

c) introducing into a plant cell, plant, or a part thereof a nucleic acid operably linked to a polynucleotide encoding a chloroplast localization sequence, wherein the nucleic acid comprises

i) the nucleic acid sequence of SEQ ID NO: 71513;

ii) a nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 71514;

iii) a nucleic acid sequence encoding an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 71514; or

iv) a nucleic acid which hybridizes with the complement of the nucleic acid of (i) under high stringent hybridization conditions of about 0.2×SSC at 50° C. to 65° C.;

wherein expression of the nucleic acid results in a decrease in the amount of verbascose relative to a corresponding control plant cell, plant or part thereof.

4. The process of claim 3 , further comprising regenerating a plant from the plant cell.

5. An expression cassette comprising a promoter that functions in plants operably linked to a nucleic acid encoding a fusion protein, wherein the fusion protein comprises a chloroplast transit peptide and a polypeptide comprising

i) an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 71514;

ii) the amino acid sequence of SEQ ID NO: 71514; or

iii) an amino acid sequence encoded by a nucleic acid molecule which hybridizes under high stringent hybridization conditions of about 0.2×SSC at 50° C. to 65° C. with the complement of a nucleic acid molecule comprising the sequence of SEQ ID NO: 71513;

wherein the expression cassette when expressed in a plant cell, plant, or part thereof results in a decrease in the amount of verbascose relative to a corresponding control plant cell, plant, or part thereof.

6. The expression cassette of claim 5 , wherein the polypeptide comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 71514.

7. The expression cassette of claim 5 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 71514.

8. A vector comprising the expression cassette of claim 5 .

9. A plant cell, plant or part thereof comprising the expression cassette of claim 6 .

10. A food or feed composition comprising the plant or part thereof of claim 9 .

11. A transgenic plant or part thereof obtained by the method of claim 3 .

12. A plant cell obtained by the method of claim 3 .

13. A transgenic seed obtained from the plant of claim 9 , wherein said transgenic seed comprises said expression cassette.

14. The process of claim 1 , further comprising regenerating a plant from the plant cell.

15. A plant cell, plant or part thereof comprising the expression cassette of claim 5 .

16. A food or feed composition comprising the plant or part thereof of claim 15 .

17. A transgenic plant cell, plant, or part thereof obtained by the method of claim 2 .

18. A plant cell, plant, or part thereof obtained by the method of claim 1 .

19. A transgenic seed obtained from the plant of claim 15 , wherein said transgenic seed comprises said expression cassette.

20. A process for producing a polypeptide in a plant cell, plant, or part thereof, comprising:

(a) obtaining a plant cell, plant, or part thereof transformed stably or transiently with a nucleic acid molecule; and

(b) producing a polypeptide encoded by said nucleic acid molecule in said plant cell, plant, or part thereof,

wherein said polypeptide is expressed in, or targeted to, a chloroplast of said plant cell, plant, or part thereof,

wherein said nucleic acid molecule is:

(i) a nucleic acid molecule encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 71514;

(ii) a nucleic acid molecule comprising the nucleic acid molecule of SEQ ID NO: 71513;

(iii) a nucleic acid molecule encoding a polypeptide having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 71514; or

(iv) a nucleic acid molecule which hybridizes with the complement of the nucleic acid molecule of (ii) under high stringent hybridization conditions of about 0.2×SSC at 50° C. to 65° C.,

and wherein expression of said nucleic acid molecule in said plant cell, plant, or part thereof confers a decrease in the amount of verbascose relative to a corresponding control plant cell, plant, or part thereof.

21. The process of claim 20 , wherein said plant cell, plant, or part thereof is transformed stably or transiently with a nucleic acid construct comprising said nucleic acid molecule.

22. The process of claim 20 , wherein said plant cell, plant, or part thereof is transformed stably or transiently with a vector comprising said nucleic acid molecule.

Assignments (1)

Priority Claims (25)

Continuity (1)