DGAT genes for increased seed storage lipid production and altered fatty acid profiles in oilseed plants
Transgenic oilseeds having increased total fatty acid content of at least 10% and altered fatty acid profiles when compared to the total fatty acid content of null segregant oilseeds are described. Novel DGAT genes are used to achieve the increase in seed storage lipids.
1. A transgenic oilseed having an increased total fatty acid content of at least 10% when compared to the total fatty acid content of a non-transgenic null segregant oilseed, wherein said transgenic oilseed comprises a recombinant construct having at least one type 1 plant DGAT sequence having 80% sequence identity to SEQ ID NO: 153 and further comprising the following amino acid substitutions:
a non-alanine at a position corresponding to position 146 of SEQ ID NO: 153 to alanine;
a non-lysine at a position corresponding to position 206 of SEQ ID NO: 153 to lysine;
a non-leucine at a position corresponding to position 273 of SEQ ID NO: 153 to leucine;
a non-valine at a position corresponding to position 364 of SEQ ID NO: 153 to valine; and
a non-valine at a position corresponding to position 483 of SEQ ID NO: 153 to valine.
2. The transgenic oilseed of claim 1 , wherein said transgenic oilseed is selected from the group consisting of soybean, corn, canola, sunflower, flax, cotton, and safflower.
3. The transgenic oilseed of claim 1 , wherein the type 1 plant DGAT sequence further comprises each of the following amino acid substitutions:
a non-methionine at a position corresponding to position 170 of SEQ ID NO: 153 to methionine;
a non-glutamic acid at a position corresponding to position 258 of SEQ ID NO: 153 to glutamic acid; and
a non-serine at a position corresponding to position 422 of SEQ ID NO: 153 to serine.
4. The transgenic oilseed of claim 1 , wherein the type 1 plant DGAT sequence further comprises each of the following amino acid substitutions:
a non-asparagine at a position corresponding to position 58 of SEQ ID NO: 153 to asparagine;
a non-phenylalanine at a position corresponding to position 231 of SEQ ID NO: 153 to phenylalanine;
a non-threonine at a position corresponding to position 264 of SEQ ID NO: 153 to threonine;
a non-valine at a position corresponding to position 303 of SEQ ID NO: 153 to valine;
a non-methionine at a position corresponding to position 440 of SEQ ID NO: 153 to methionine; and
a non-lysine at a position corresponding to position 467 of SEQ ID NO: 153 to lysine.
5. The transgenic oilseed of claim 4 , wherein the type 1 plant DGAT sequence further comprises each of the following amino acid substitutions:
a non-methionine at a position corresponding to position 170 of SEQ ID NO: 153 to methionine;
a non-glutamic acid at a position corresponding to position 258 of SEQ ID NO: 153 to glutamic acid; and
a non-serine at a position corresponding to position 422 of SEQ ID NO: 153 to serine.
6. The transgenic oilseed of claim 1 , wherein said type 1 plant DGAT comprises at least 90% sequence identity to SEQ ID NO: 153.
7. The transgenic oilseed of claim 1 , wherein said type 1 plant DGAT comprises at least 95% sequence identity to SEQ ID NO: 153.
8. A method of making a transgenic oilseed having an increased total fatty acid content of at least 10% when compared to the total fatty acid content of a non-transgenic null segregant oilseed, said method comprising:
(a) transforming at least one oilseed cell with a recombinant construct having at least one type 1 plant DGAT sequence having 80% sequence identity to SEQ ID NO: 153 and further comprising the following amino acid substitutions:
a non-alanine at a position corresponding to position 146 of SEQ ID NO: 153 to alanine;
a non-lysine at a position corresponding to position 206 of SEQ ID NO: 153 to lysine;
a non-leucine at a position corresponding to position 273 of SEQ ID NO: 153 to leucine;
a non-valine at a position corresponding to position 364 of SEQ ID NO: 153 to valine; and
a non-valine at a position corresponding to position 483 of SEQ ID NO: 153 to valine;
(b) selecting the transformed oilseed cell(s) of step (a) having an increased total fatty acid content of at least 10% when compared to the fatty acid content of a non-transgenic null segregant oilseed cell; and,
(c) regenerating a transgenic oilseed plant from said transformed oilseed cell(s).
9. The method of claim 8 , wherein said transgenic oilseed is selected from the group consisting of soybean, corn, canola, sunflower, flax, cotton, and safflower.
10. The method of claim 8 , wherein the recombinant construct having at least one type 1 plant DGAT sequence further comprises each of the following amino acid substitutions:
a non-methionine at a position corresponding to position 170 of SEQ ID NO: 153 to methionine;
a non-glutamic acid at a position corresponding to position 258 of SEQ ID NO: 153 to glutamic acid; and
a non-serine at a position corresponding to position 422 of SEQ ID NO: 153 to serine.
11. The method of claim 8 , wherein the recombinant construct having at least one type 1 plant DGAT sequence further comprises each of the following amino acid substitutions:
a non-asparagine at a position corresponding to position 58 of SEQ ID NO: 153 to asparagine;
a non-phenylalanine at a position corresponding to position 231 of SEQ ID NO: 153 to phenylalanine;
a non-threonine at a position corresponding to position 264 of SEQ ID NO: 153 to threonine;
a non-valine at a position corresponding to position 303 of SEQ ID NO: 153 to valine;
a non-methionine at a position corresponding to position 440 of SEQ ID NO: 153 to methionine; and
a non-lysine at a position corresponding to position 467 of SEQ ID NO: 153 to lysine.
12. The method of claim 11 , wherein the recombinant construct having at least one type 1 plant DGAT sequence further comprises each of the following amino acid substitutions:
a non-methionine at a position corresponding to position 170 of SEQ ID NO: 153 to methionine;
a non-glutamic acid at a position corresponding to position 258 of SEQ ID NO: 153 to glutamic acid; and
a non-serine at a position corresponding to position 422 of SEQ ID NO: 153 to serine.
13. A method of producing a food or feed product, said method comprising:
(a) obtaining a transgenic seed from a transgenic oilseed, wherein said transgenic oilseed comprises a recombinant construct having at least one type 1 plant DGAT sequence having 80% sequence identity to SEQ ID NO: 153 and further comprising the following amino acid substitutions:
a non-alanine at a position corresponding to position 146 of SEQ ID NO: 153 to alanine;
a non-lysine at a position corresponding to position 206 of SEQ ID NO: 153 to lysine;
a non-leucine at a position corresponding to position 273 of SEQ ID NO: 153 to leucine;
a non-valine at a position corresponding to position 364 of SEQ ID NO: 153 to valine; and
a non-valine at a position corresponding to position 483 of SEQ ID NO: 153 to valine,
and wherein said transgenic seed has an increased total fatty acid content of at least 10% when compared to the total fatty acid content of a non-transgenic null segregant seed; and
(b) processing said seed to produce a food or feed product.
14. The method of claim 13 , wherein said oilseed is selected from the group consisting of soybean, corn, canola, sunflower, flax, cotton, and safflower.
15. The method of claim 13 , wherein type 1 plant DGAT sequence further comprises each of the following amino acid substitutions:
a non-methionine at a position corresponding to position 170 of SEQ ID NO: 153 to methionine;
a non-glutamic acid at a position corresponding to position 258 of SEQ ID NO: 153 to glutamic acid; and
a non-serine at a position corresponding to position 422 of SEQ ID NO: 153 to serine.
16. The method of claim 13 , wherein type 1 plant DGAT sequence further comprises each of the following amino acid substitutions:
a non-asparagine at a position corresponding to position 58 of SEQ ID NO: 153 to asparagine;
a non-phenylalanine at a position corresponding to position 231 of SEQ ID NO: 153 to phenylalanine;
a non-threonine at a position corresponding to position 264 of SEQ ID NO: 153 to threonine;
a non-valine at a position corresponding to position 303 of SEQ ID NO: 153 to valine;
a non-methionine at a position corresponding to position 440 of SEQ ID NO: 153 to methionine; and
a non-lysine at a position corresponding to position 467 of SEQ ID NO: 153 to lysine.
17. The method of claim 16 , wherein type 1 plant DGAT sequence further comprises each of the following amino acid substitutions:
a non-methionine at a position corresponding to position 170 of SEQ ID NO: 153 to methionine;
a non-glutamic acid at a position corresponding to position 258 of SEQ ID NO: 153 to glutamic acid; and
a non-serine at a position corresponding to position 422 of SEQ ID NO: 153 to serine.