Synthetic brassica-derived chloroplast transit peptides

Patent metadata

Field	Value
Publication number	US-9540654-B2
Application number	US-201313757435-A
Country	US
Kind code	B2
Filing date	Feb 1, 2013
Priority date	Feb 1, 2012
Publication date	Jan 10, 2017
Grant date	Jan 10, 2017

Abstract

This disclosure concerns compositions and methods for targeting peptides, polypeptides, and proteins to plastids of plastid-containing cells. In some embodiments, the disclosure concerns chloroplast transit peptides that may direct a polypeptide to a plastid, and nucleic acid molecules encoding the same. In some embodiments, the disclosure concerns methods for producing a transgenic plant material (e.g., a transgenic plant) comprising a chloroplast transit peptide, as well as plant materials produced by such methods, and plant commodity products produced therefrom.

First claim

What may be claimed is:

1. An isolated nucleic acid molecule comprising:
a first nucleotide sequence that encodes a peptide that is at least 98% identical to SEQ ID NO:6 or SEQ ID NO:8; and
a heterologous nucleotide sequence encoding a peptide of interest that is operably linked to the first nucleotide sequence.

2. The isolated nucleic acid molecule of claim 1 , wherein the peptide is selected from the group consisting of SEQ ID NOs:6 and 8.

3. The isolated nucleic acid molecule of claim 1 , further comprising at least one additional nucleotide sequence(s), each encoding a chloroplast transit peptide, wherein the additional nucleotide sequence(s) are operably linked to the heterologous nucleotide sequence.

4. The isolated nucleic acid molecule of claim 3 , wherein the at least one additional nucleotide sequence(s) is from an organism selected from the group consisting of prokaryotes, lower photosynthetic eukaryotes, and Chlorophytes.

5. The isolated nucleic acid molecule of claim 1 , wherein the first nucleotide sequence and the heterologous nucleotide sequence are operably linked to one or more regulatory sequences.

6. A chimeric polypeptide encoded by the nucleic acid molecule of claim 1 .

7. The chimeric polypeptide of claim 6 , wherein the peptide of interest is a biologically-active peptide.

8. The chimeric polypeptide of claim 6 , wherein the peptide of interest is a fluorescent peptide.

9. The chimeric polypeptide of claim 7 , wherein the biologically-active peptide is an enzyme.

10. The chimeric polypeptide of claim 7 , wherein the biologically-active peptide is normally expressed in a plastid of a cell wherein the peptide is natively expressed.

11. The chimeric polypeptide of claim 7 , wherein the biologically-active peptide is involved in a process selected from the group consisting of virus resistance, bacterial pathogen resistance, insect resistance, nematode resistance, fungal resistance, plant vigor, plant yield, temperature tolerance, soil condition tolerance, low light level tolerance, low water level tolerance, high water level tolerance, chemical environment tolerance, seed color, starch modification, amino acid synthesis, photosynthesis, synthesis of fatty acids, oil synthesis, synthesis of arotenoids, synthesis of terpenoids, synthesis of starch, and herbicide resistance.

12. The chimeric polypeptide of claim 7 , wherein the biologically-active peptide is selected from the group consisting of zeaxanthin epoxidase, choline monooxygenase, ferrochelatase, omega-3 fatty acid desaturase, glutamine synthetase, provitamin A, hormones, Bt toxin proteins, and markers useful in identification of plants comprising a trait of interest.

13. The chimeric polypeptide of claim 11 , wherein the biologically-active peptide is involved in herbicide resistance.

14. The chimeric polypeptide of claim 9 , wherein the biologically-active peptide is selected from the group consisting of: acetolactase synthase (ALS), mutated ALS, precursors of ALS, 3-enolpyruvylshikimate-5-phosphate synthetase (EPSPS), CP4 EPSPS, a class III EPSPS, and glutamine synthetase.

15. The nucleic acid molecule of claim 1 , wherein the molecule is a plant expression vector.

16. A plant material comprising the nucleic acid molecule of claim 1 .

17. The plant material of claim 16 , wherein the plant material is selected from the group consisting of a plant cell, a plant tissue, a plant tissue culture, a callus culture, a plant part, and a whole plant.

18. The plant material of claim 16 , further comprising a polypeptide comprising the peptide of interest.

19. The plant material of claim 18 , wherein the peptide of interest is a portion of the polypeptide that is targeted to a plastid in a cell of the plant material.

20. The plant material of claim 16 , wherein the nucleic acid molecule is stably integrated into the genome of a cell from the plant material.

21. The plant material of claim 16 , wherein the plant material is a whole plant.

22. The plant material of claim 16 , wherein the plant material is from a plant selected from the group consisting of Arabidopsis , alfalfa, Brassica , beans, broccoli, cabbage, carrot, cauliflower, celery, Chinese cabbage, cotton, cucumber, eggplant, lettuce, melon, pea, pepper, peanut, potato, pumpkin, radish, rapeseed, spinach, soybean, squash, sugarbeet, sunflower, tobacco, tomato, watermelon, corn, onion, rice, sorghum, wheat, rye, millet, sugarcane, oat, triticale, switchgrass, and turfgrass.

23. A method for producing a transgenic plant material, the method comprising
transforming a plant material with the nucleic acid molecule of claim 1 .

24. The method according to claim 23 , wherein the plant material is selected from the group consisting of a plant cell, a plant tissue, a plant tissue culture, a callus culture, a plant part, and a whole plant.

25. The method according to claim 23 , wherein the plant material is not a whole plant.

26. A transgenic plant material produced by the method according to claim 23 .

27. A transgenic plant regenerated from the transgenic plant material of claim 26 .

28. A transgenic plant commodity product produced from the plant material of claim 26 .

29. The transgenic plant material of claim 26 , wherein the peptide of interest is a biologically-active peptide.

30. The transgenic plant material of claim 29 , wherein the biologically-active peptide is involved in a process selected from the group consisting of virus resistance, bacterial pathogen resistance, insect resistance, nematode resistance, fungal resistance, plant vigor, plant yield, temperature tolerance, soil condition tolerance, low light level tolerance, low water level tolerance, high water level tolerance, chemical environment tolerance, seed color, starch modification, amino acid synthesis, photosynthesis, synthesis of fatty acids, oil synthesis, synthesis of arotenoids, synthesis of terpenoids, synthesis of starch, and herbicide resistance.

31. The transgenic plant material of claim 29 , wherein the biologically-active peptide is selected from the group consisting of zeaxanthin epoxidase, choline monooxygenase, ferrochelatase, omega-3 fatty acid desaturase, glutamine synthetase, provitamin A, hormones, Bt toxin proteins, and markers useful in identification of plants comprising a trait of interest.

32. The transgenic plant material of claim 30 , wherein the biologically-active peptide is involved in herbicide resistance.

33. The transgenic plant material of claim 29 , wherein the biologically-active peptide is selected from the group consisting of acetolactase synthase (ALS), mutated ALS, precursors of ALS, 3-enolpyruvylshikimate-5-phosphate synthetase (EPSPS), CP4 EPSPS, a class III EPSPS, and glutamine synthetase.

34. The transgenic plant material of claim 32 , wherein the plant material exhibits increased herbicide resistance or herbicide tolerance when compared to a wild-type plant material of the same species.

35. The plant material of claim 17 , wherein the plant material is a plant cell that is incapable of regeneration to produce a plant.

36. The method of claim 24 , wherein the plant material is a plant cell that is incapable of regeneration to produce a plant.

Assignees

• Dow Agrosciences Llc

Inventors

• Lira Justin M
• Cicchillo Robert M
• Yerkes Carla N
• Robinson Andrew E

Classifications

• C12N15/8275

  Glyphosate · CPC title

• A01H5/10

  Seeds · CPC title

• C12N9/0071

  acting on paired donors with incorporation of molecular oxygen (1.14) · CPC title

• C12N9/96

  Stabilising an enzyme by forming an adduct or a composition; Forming enzyme conjugates · CPC title

• C12Y114/11

  with 2-oxoglutarate as one donor, and incorporation of one atom each of oxygen into both donors (1.14.11) · CPC title