Methods and compositions for short stature plants through manipulation of gibberellin metabolism to increase harvestable yield

Patent metadata

Field	Value
Publication number	US-12319922-B2
Application number	US-202217713344-A
Country	US
Kind code	B2
Filing date	Apr 5, 2022
Priority date	Aug 17, 2016
Publication date	Jun 3, 2025
Grant date	Jun 3, 2025

Abstract

The present disclosure provides compositions and methods for altering gibberellin (GA) content in corn or other cereal plants. Methods and compositions are also provided for altering the expression of genes related to gibberellin biosynthesis through suppression, mutagenesis and/or editing of specific subtypes of GA20 or GA3 oxidase genes. Modified plant cells and plants having a suppression element or mutation reducing the expression or activity of a GA oxidase gene are further provided comprising reduced gibberellin levels and improved characteristics, such as reduced plant height and increased lodging resistance, but without off-types.

First claim

What is claimed is:

1. A recombinant DNA construct comprising a transcribable DNA sequence encoding a non-coding RNA molecule, wherein the non-coding RNA molecule comprises a targeting sequence that is:
(a) at least 80% complementary to at least 19 consecutive nucleotides of a first mRNA molecule encoding a first endogenous gibberellin (GA) oxidase protein in a corn plant, the first endogenous GA oxidase protein being at least 90% identical to SEQ ID NO: 9; and
(b) at least 80% complementary to at least 19 consecutive nucleotides of a second mRNA molecule encoding a second endogenous GA oxidase protein in a corn plant, the second endogenous GA oxidase protein being at least 90% identical to SEQ ID NO: 15;

wherein the transcribable DNA sequence is operably linked to a leaf promoter.

2. The recombinant DNA construct of claim 1 , wherein the non-coding RNA molecule reduces the expression levels of the first and second mRNA molecules in at least one tissue of a transgenic corn plant comprising the recombinant DNA construct, relative to a control plant, when the non-coding RNA molecule is expressed in the transgenic corn plant.

3. The recombinant DNA construct of claim 1 , wherein the non-coding RNA molecule reduces the expression levels of the first and second endogenous GA oxidase proteins in at least one tissue of a transgenic corn plant comprising the recombinant DNA construct, relative to a control plant, when the non-coding RNA molecule is expressed in the transgenic corn plant.

4. The recombinant DNA construct of claim 1 , wherein the targeting sequence of the non-coding RNA molecule is at least 90% complementary to at least 19 consecutive nucleotides of a sequence selected from the group consisting of SEQ ID NOs: 7, 8, 13, and 14.

5. The recombinant DNA construct of claim 1 , wherein the targeting sequence of the non-coding RNA molecule is at least 90% complementary to at least 19 consecutive nucleotides of the first mRNA molecule encoding the first endogenous GA20 oxidase protein.

6. The recombinant DNA construct of claim 3 , wherein the targeting sequence of the non-coding RNA molecule is 100% complementary to at least 19 consecutive nucleotides of the first mRNA molecule encoding the first endogenous GA20 oxidase protein.

7. The recombinant DNA construct of claim 3 , wherein the targeting sequence of the non-coding RNA molecule is at least 90% complementary to at least 19 consecutive nucleotides of SEQ ID NO: 7 or 8.

8. The recombinant DNA construct of claim 1 , wherein the targeting sequence of the non-coding RNA molecule is at least 90% complementary to at least 19 consecutive nucleotides of the second mRNA molecule encoding the second endogenous GA20 oxidase protein.

9. The recombinant DNA construct of claim 6 , wherein the targeting sequence of the non-coding RNA molecule is 100% complementary to at least 19 consecutive nucleotides of the second mRNA molecule encoding the second endogenous GA20 oxidase protein.

10. The recombinant DNA construct of claim 6 , wherein the targeting sequence of the non-coding RNA molecule is at least 90% complementary to at least 19 consecutive nucleotides of SEQ ID NO: 13 or 14.

11. The recombinant DNA construct of claim 1 , wherein the targeting sequence of the non-coding RNA molecule is at least 90% complementary to at least 21 consecutive nucleotides of the first mRNA molecule encoding the first endogenous GA20 oxidase protein, and is at least 90% complementary to at least 21 consecutive nucleotides of the second mRNA molecule encoding the second GA20 oxidase protein.

12. The recombinant DNA construct of claim 1 , wherein the targeting sequence of the non-coding RNA molecule is 100% complementary to at least 21 consecutive nucleotides of the first mRNA molecule encoding the first endogenous GA20 oxidase protein, and is 100% complementary to at least 21 consecutive nucleotides of the second mRNA molecule encoding the second GA20 oxidase protein.

13. The recombinant DNA construct of claim 1 , wherein the non-coding RNA molecule encoded by the transcribable DNA sequence is a precursor miRNA or siRNA that is processed or cleaved in a plant cell to form a mature miRNA or siRNA.

14. The recombinant DNA construct of claim 1 , wherein the leaf promoter is selected from the group consisting of: a corn pyruvate phosphate dikinase promoter, a corn fructose 1,6 bisphosphate aldolase promoter, a rice Nadh-Gogat promoter, a ribulose biphosphate carboxylase (RuBisCO) promoter, a RuBisCO small subunit promoter, a chlorophyll a/b binding protein gene promoter, a phosphoenolpyruvate carboxylase promoter, and a Myb gene promoter.

15. The recombinant DNA construct of claim 1 , wherein the leaf promoter comprises a DNA sequence that is at least 80% identical to SEQ ID NO: 72, SEQ ID NO: 73, or SEQ ID NO: 74.

16. The recombinant DNA construct of claim 1 , wherein the leaf promoter comprises a DNA sequence that is at least 90% identical to SEQ ID NO: 72, SEQ ID NO: 73, or SEQ ID NO: 74.

17. The recombinant DNA construct of claim 1 , wherein the leaf promoter comprises a DNA sequence that is 100% identical to SEQ ID NO: 72, SEQ ID NO: 73, or SEQ ID NO: 74.

18. A transgenic corn plant comprising a recombinant DNA construct comprising a transcribable DNA sequence encoding a non-coding RNA molecule, wherein the non-coding RNA molecule comprises a targeting sequence that is:
(a) at least 80% complementary to at least 19 consecutive nucleotides of a first mRNA molecule encoding a first endogenous GA oxidase protein in a corn plant, the first endogenous GA oxidase protein being at least 90% identical to SEQ ID NO: 9; and
(b) at least 80% complementary to at least 19 consecutive nucleotides of a second mRNA molecule encoding a second endogenous GA oxidase protein in a corn plant, the second endogenous GA oxidase protein being at least 90% identical to SEQ ID NO: 15;

wherein the transcribable DNA sequence is operably linked to a leaf promoter.

19. The transgenic corn plant of claim 18 , wherein the expression levels of the first and second mRNA molecules are reduced in at least one tissue of the transgenic corn plant relative to a control plant.

20. The transgenic corn plant of claim 18 , wherein the expression levels of the first and second endogenous GA oxidase proteins in at least one tissue of the transgenic corn plant relative to a control plant.

21. The transgenic corn plant of claim 18 , wherein the transgenic corn plant has a shorter plant height relative to a control plant.

22. The transgenic corn plant of claim 18 , wherein the height of the transgenic plant is at least 10% shorter than a control plant.

23. The transgenic corn plant of claim 18 , wherein the height of the transgenic plant is at least 20% shorter than a control plant.

24. The transgenic corn plant of claim 18 , wherein the height of the transgenic plant is at least 30% shorter than a control plant.

25. The transgenic corn plant of claim 18 , wherein the height of the transgenic plant is at least 40% shorter than a control plant.

26. The transgenic corn plant of claim 18 , wherein expression of the non-coding RNA molecule in the corn plant reduces the level of one or more active GAs in the corn plant as compared to a control plant.

27. The transgenic corn plant of claim 18 , wherein the level of one or more active GAs in at least one internode tissue of the stem or stalk of the transgenic

Assignees

• Monsanto Technology Llc

Inventors

• Allen Edwards M
• Boddu Jayanand
• Dietrich Charles R
• Goldshmidt Alexander
• Howell Miya
• Kosola Kevin R
• Manjunath Sivalinganna
• Neelam Anil
• Rymarquis Linda
• Slewinski Thomas L
• Venkatesh Tyamagondlu V
• Wang Huai

Classifications

• C12N15/8297 Primary

  Gibberellins; GA3 · CPC title

• C12N15/8273 Primary

  for drought, cold, salt resistance · CPC title

• C12N15/1137

  against enzymes (viral enzymes C12N15/1131; receptors C12N15/1138) · CPC title

• C12N2310/20

  involving clustered regularly interspaced short palindromic repeats [CRISPR] · CPC title

• A01H6/4684

  Zea mays [maize] · CPC title