Use of a seed specific promoter to drive ODP1 expression in cruciferous oilseed plants to increase oil content while maintaining normal germination

Patent metadata

Field	Value
Publication number	US-9284571-B2
Application number	US-201313769539-A
Country	US
Kind code	B2
Filing date	Feb 18, 2013
Priority date	Apr 1, 2009
Publication date	Mar 15, 2016
Grant date	Mar 15, 2016

Abstract

A recombinant DNA construct comprising a polynucleotide encoding an ODP1 polypeptide operably linked to a sucrose synthase 2 promoter where this construct can be used to increase oil content in the seeds of a cruciferous oilseed plant while maintaining normal germination is disclosed. A method for increasing oil content in the seeds of a cruciferous oilseed plant while maintaining normal germination using this construct is also disclosed.

First claim

What is claimed is:

1. A recombinant DNA construct comprising a polynucleotide encoding an ODP1 polypeptide operably linked to a sucrose synthase 2 promoter (SUS2) wherein the SUS2 promoter comprises a nucleotide sequence having at least 95% sequence identity to SEQ ID NO:43, wherein said nucleotide sequence has seed specific promoter activity and wherein the amino acid sequence of said ODP1 polypeptide has at least 90% sequence identity to SEQ ID NO:39 and comprises two APETALA2 (AP2) domains and wherein expression of said ODP1 polypeptide increases oil content in the seeds of a cruciferous oilseed plant while maintaining normal germination.

2. The recombinant DNA construct of claim 1 wherein the amino acid sequence of said ODP1 polypeptide has at least 95% sequence identity to SEQ ID NO:39.

3. The recombinant DNA construct of claim 1 wherein the amino acid sequence of said ODP1 polypeptide comprises SEQ ID NO:39.

4. The recombinant DNA construct of claim 1 wherein the sucrose synthase 2 promoter comprises the nucleotide sequence of SEQ ID NO:43.

5. The recombinant DNA construct of claim 2 , wherein the sucrose synthase 2 promoter comprises the nucleotide sequence of SEQ ID NO:43.

6. The recombinant DNA construct of claim 3 , wherein the sucrose synthase 2 promoter comprises the nucleotide sequence of SEQ ID NO:43.

7. The recombinant DNA construct of claim 1 wherein the oilseed plant is canola or Arabidopsis.

8. A transgenic cruciferous oilseed plant comprising in its genome the recombinant DNA construct of claim 1 .

9. The transgenic cruciferous oilseed plant of claim 8 wherein the cruciferous oilseed plant is selected from the group consisting of canola and Arabidopsis.

10. A transgenic seed obtained from the plant of claim 8 , wherein said seed comprises in its genome said recombinant DNA construct.

11. The transgenic cruciferous oilseed plant of claim 8 , wherein the amino acid sequence of said ODP1 polypeptide comprises SEQ ID NO:39.

12. The transgenic cruciferous oilseed plant of claim 8 , wherein the sucrose synthase 2 promoter comprises the nucleotide sequence of SEQ ID NO:43 and wherein the amino acid sequence of said ODP1 polypeptide has at least 95% sequence identity to SEQ ID NO:39.

13. The transgenic cruciferous oilseed plant of claim 8 , wherein the sucrose synthase 2 promoter comprises the nucleotide sequence of SEQ ID NO:43 and wherein the amino acid sequence of said ODP1 polypeptide comprises SEQ ID NO:39.

14. A transgenic seed obtained from the plant of claim 8 , wherein said seed comprises in its genome said recombinant DNA construct and wherein the amino acid sequence of said ODP1 polypeptide comprises SEQ ID NO:39.

15. A transgenic seed obtained from the plant of claim 8 , wherein said seed comprises in its genome said recombinant DNA construct and wherein the sucrose synthase 2 promoter comprises the nucleotide sequence of SEQ ID NO:43 and wherein the amino acid sequence of ODP1 polypeptide has at least 95% sequence identity to SEQ ID NO:39.

16. A transgenic seed obtained from the plant of claim 8 , wherein said seed comprises in its genome said recombinant DNA construct and wherein the sucrose synthase 2 promoter comprises the nucleotide sequence of SEQ ID NO:43 and wherein the amino acid sequence of said ODP1 polypeptide comprises SEQ ID NO:39.

17. A method for producing a transgenic cruciferous oilseed plant comprising transforming a cruciferous oilseed plant cell with the recombinant DNA construct of claim 1 and regenerating a transgenic cruciferous oilseed plant from the transformed cruciferous oilseed plant cell, wherein the transgenic cruciferous oilseed plant comprises in its genome said recombinant DNA construct.

18. The method of claim 17 wherein the cruciferous oilseed plant is selected from the group consisting of canola and Arabidopsis .

19. A method for increasing oil content in seeds of a transgenic cruciferous oilseed plant while maintaining normal germination, said method comprising:
(a) transforming a cruciferous oilseed plant cell with a recombinant DNA construct comprising a polynucleotide encoding an ODP1 polypeptide, wherein the amino acid sequence of said ODP1 polypeptide has at least 90% sequence identity to SEQ ID NO:39 and comprises two APETALA2 (AP2) domains, said polynucleotide being operably linked to a promoter having a nucleotide sequence at least 95% identical to SEQ ID NO: 43, wherein said nucleotide sequence has seed specific promoter activity;
(b) regenerating a transgenic cruciferous oilseed plant from the transformed cell of step (a), wherein said plant comprises the recombinant DNA construct;
(c) obtaining a transgenic progeny plant derived from the transgenic cruciferous oilseed plant of step (b), wherein the transgenic progeny plant comprises in its genome the recombinant DNA construct;
(d) assaying the transgenic progeny plant obtained from step (c) for oil level and germination; and
(e) selecting those transgenic progeny plants having seeds comprising said recombinant DNA construct and having an increased level of oil and normal germination when compared to seeds obtained from a control cruciferous oilseed plant, wherein said control plant does not comprise the recombinant DNA construct.

20. The method of claim 19 wherein the amino acid sequence of the ODP1 polypeptide comprises the sequence of SEQ ID NO:39.

21. The method of claim 19 wherein the promoter comprises SEQ ID NO:43.

22. The method of claim 21 wherein the ODP1 polypeptide comprises at least 95% sequence identity to SEQ ID NO: 39.

23. The method of claim 19 wherein the cruciferous oilseed plant is canola or Arabidopsis.

Assignees

• Du Pont
• Du Pont

Inventors

• Damude Howard Glenn
• Everard John D
• Meyer Knut
• Ripp Kevin G
• Stecca Kevin L

Classifications

• C12N15/8247 Primary

  involving modified lipid metabolism, e.g. seed oil composition · CPC title

• C12N15/8201

  Methods for introducing genetic material into plant cells, e.g. DNA, RNA, stable or transient incorporation, tissue culture methods adapted for transformation · CPC title