METHODS OF INCREASING SEED YIELD

• US 20200131524A1
• Filed: 01/10/2018
• Published: 04/30/2020
• Est. Priority Date: 01/10/2017
• Status: Active Grant

• Current Assignee: Plant Bioscience Limited (Alnylam Pharmaceuticals Incorporated)
• Original Assignee: Plant Bioscience Limited (Alnylam Pharmaceuticals Incorporated)
• Inventors: Miller, Charlotte, Bevan, Michael
• Granted Patent: US 11,473,097 B2
• Time in Patent Office: Days
• CPC Class Codes: C12N 15/8213 Targeted insertion of genes into the plant genome by homologous recombination
• CPC Class Codes: C12N 15/8218 Antisense, co-suppression, viral induced gene silencing [VIGS], post-transcriptional induced gene silencing [PTGS]
• CPC Class Codes: C12N 15/8242 with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits
• CPC Class Codes: C12N 15/8245 involving modified carbohydrate or sugar alcohol metabolism, e.g. starch biosynthesis
• CPC Class Codes: C12N 15/8247 involving modified lipid metabolism, e.g. seed oil composition
• CPC Class Codes: C12N 15/8251 Amino acid content, e.g. synthetic storage proteins, altering amino acid biosynthesis
• CPC Class Codes: C12N 15/8261 with agronomic (input) traits, e.g. crop yield
• CPC Class Codes: C12Q 1/6895 for plants, fungi or algae
• CPC Class Codes: C12Q 2600/13 Plant traits
• CPC Class Codes: Y02A 40/146 Genetically Modified [GMO] plants, e.g. transgenic plants

Abstract

The invention relates to methods for increasing seed yield, increasing the total content of protein and/or lipid in seeds and reducing glucosinolate levels by reducing the expression or activity of UPL3. The invention also relates to genetically altered plants characterised by the above phenotypes and methods of producing such plants.

Claims

1. A method of increasing seed yield or lipid and/or protein content in a plant seed or reducing the levels of glucosinolate in a plant, the method comprising reducing or abolishing the expression of at least one nucleic acid encoding a ubiquitin protein ligase 3 (UPL3) polypeptide and/or reducing the activity of a UPL3 polypeptide in said plant.

2. (canceled)

3. (canceled)

4. (canceled)

5. The method of claim 1, wherein the method comprises introducing at least one mutation into the nucleic acid sequence encoding UPL3 or the promoter of said UPL3.

6. (canceled)

7. (canceled)

8. The method of claim 5, wherein the UPL3 nucleic acid encoding a UPL3 polypeptide comprises or consists of SEQ ID NO:

4 or 5 or a functional variant or homolog thereof, and wherein the promoter of said UPL3 nucleic acid sequence comprises or consists of SEQ ID NO;

1 or 2 or a functional variant or homolog thereof.

9. The method of claim 5, wherein the mutation is introduced using targeted genome modification, preferably ZFNs, TALENs or CRISPR/Cas9 or wherein the mutation is introduced using mutagenesis, preferably TILLING or T-DNA insertion.

10. (canceled)

11. The method of claim 1, the method comprising using RNA interference to reduce or abolish the expression of a UPL3 nucleic acid.

12. (canceled)

13. The method of claim 1, wherein the plant is selected from a dicot plant.

14. The plant of claim 13, wherein the plant is selected from any dicotyledonous oilseed crop, such as Brassica oilseed crops such as B. juncea, soybean, sunflower, linseed, cotton, hemp, oilpalm, coconut, peanut, safflower, Camelina and olive or wherein the plant is selected from B. oleracea, maize, rice, wheat or barley.

15. The method of claim 14, wherein the plant is Brassica napus.

16. A genetically altered plant, part thereof or plant cell, wherein said plant comprises at least one mutation into at least one ubiquitin protein ligase 3 (UPL3) gene and/or at least one mutation in the UPL3 promoter.

17. The plant of claim 16, wherein said plant is characterised by an increase in seed yield or an increase in seed protein and/or lipid content or reduced glucosinolate levels compared to a wild-type or control plant.

18. (canceled)

19. (canceled)

20. (canceled)

21. (canceled)

22. (canceled)

23. The plant of claim 16, wherein the UPL3 gene comprises or consists of a nucleic acid sequence as defined in any of SEQ ID NOs:

4 to 7 or a functional variant or homolog thereof, and wherein the promoter of the UPL3 gene comprises or consists of a nucleic acid sequence as defined in SEQ ID NO;

1 or 2 or a functional variant or homolog thereof.

24. The plant of claim 16, wherein said plant comprises an RNA interference construct that reduces the expression of a UPL3 nucleic acid.

25. The plant of claim 16, wherein the plant is a dicot.

26. The plant of claim 25, wherein the plant is selected from any dicotyledonous oilseed crop, such as Brassica oilseed crops such as B. juncea, soybean, sunflower, linseed, cotton, hemp, oilpalm, coconut, peanut, safflower, Camelina and olive or wherein the plant is selected from B. oleracea, maize, rice, wheat or barley.

27. The plant of claim 26, wherein the plant is Brassica napus.

28. The plant part of claim 16, wherein the plant part is a seed.

29. (canceled)

30. (canceled)

31. (canceled)

32. (canceled)

33. (canceled)

34. (canceled)

35. (canceled)

36. (canceled)

37. (canceled)

38. (canceled)

39. (canceled)

40. (canceled)

41. (canceled)

42. A nucleic acid construct comprising a nucleic acid sequence as defined in any one of SEQ ID NOs:

11 to 14 or a functional variant or homolog thereof, wherein said sequence is operably linked to a regulatory sequence, wherein preferably said regulatory sequence is a tissue-specific promoter.

43. (canceled)

44. (canceled)

45. (canceled)

46. A method of increasing seed yield and/or increasing seed protein and/or lipid content and/or reducing glucosinolate levels, the method comprising introducing and expressing in said plant the nucleic acid construct of claim 42.

47. (canceled)

48. (canceled)

49. A method for screening a population of plants and identifying and/or selecting a plant that has or will have reduced UPL3 expression, an increased seed yield phenotype, increased seed protein and/or lipid levels and/or reduced glucosinolate levels, preferably compared to a wild-type or control plant, the method comprising detecting in the plant or plant germplasm at least one polymorphism in the UPL3 gene and/or promoter and/or at least one polymorphism in SEQ ID NO:

58, and selecting said plant or progeny thereof.

50. (canceled)

51. The method of claim 49, wherein the method further comprises introgressing the chromosomal region comprising at least one polymorphism in the UPL3 gene or promoter into a second plant or plant germplasm to produce an introgressed plant or plant germplasm