Materials and methods for increasing the tocopherol content in seed oil

Patent metadata

Field	Value
Publication number	US-10829775-B2
Application number	US-201916405440-A
Country	US
Kind code	B2
Filing date	May 7, 2019
Priority date	Nov 14, 2014
Publication date	Nov 10, 2020
Grant date	Nov 10, 2020

Abstract

The present invention relates generally to the field of molecular biology and concerns increasing the tocopherol content of a plant relative to a control plant, comprising expressing in a plant at least one polynucleotide encoding a delta-12-desaturase, at least one polynucleotide encoding a delta-6-desaturase, at least one polynucleotide encoding a delta-6-elongase, and at least one polynucleotide encoding a delta-5-desaturase. The present invention also relates to methods for the manufacture of oil, fatty acid- or lipids-containing compositions, and to such oils and lipids as such.

First claim

The invention claimed is:

1. A transgenic seed of a transgenic Brassica plant, the transgenic seed comprising tocopherol and n-3 very long chain polyunsaturated fatty acids (VLC-PUFAs), wherein the transgenic seed has increased total tocopherol content and increased n-3 VLC-PUFAs content as compared to a seed from a wild-type control Brassica plant of the same species and variety, wherein the increased n-3 VLC-PUFAs content comprises EPA and/or DHA; wherein the increased total tocopherol content and the increased n=3 VLC-PUFAs content in the transgenic seed of the transgenic Brassica plant are positively correlated, wherein the transgenic seed comprises transgenes which comprise at least one recombinant polynucleotide encoding a delta-12-desaturase from Phythophthora species, at least one recombinant polynucleotide encoding a delta-6-desaturase from Ostreococcus species, at least one recombinant polynucleotide encoding a delta-6-elongase from Physcomitrella species or Thalassiosira species, at least one recombinant polynucleotide encoding a delta-5-desaturase from Thraustochytrium species, at least one recombinant polynucleotide encoding a delta-5-elongase from Ostreococcus species, at least one polynucleotide encoding a delta-4-desaturase from Thraustochytrium species or Pavlova species, and at least one recombinant polynucleotide encoding an omega-3-desaturase from Phythophthora species or Pythium species, and wherein expression of said transgenes in said transgenic seed results in said increase in n-3 VLC-PUFAs content and total tocopherol content in said transgenic seed.

2. The transgenic seed of claim 1 , wherein the total tocopherol content of the transgenic seed is more than 35 mg/100 g transgenic seed.

3. The transgenic seed of claim 1 , wherein the total tocopherol content of the transgenic seed is from about 36 mg/100 g transgenic seed to about 44 mg/100 g transgenic seed.

4. The transgenic seed of claim 1 , wherein the content of VLC-PUFAs in the transgenic seed is increased by at least 5%, at least 10%, at least 15%, at least 20%, or at least 30% by the transgenic seed weight as compared to a seed of a wild-type Brassica control plant of the same species and variety.

5. The transgenic seed of claim 1 , wherein the content of VLC-PUFAs in the transgenic seed is increased by 5% to 30% by the transgenic seed weight compared to a seed of a wild-type Brassica control plant of the same species and variety.

6. The transgenic seed of claim 1 , wherein the transgenic seed is from a Brassica plant of the species Brassica napus, Brassica carinata, Brassica juncea, Brassica oleracea, Brassica nigra , or Brassica rapa.

7. The transgenic seed of claim 1 , wherein the total tocopherol content is measured as the numeric average of transgenic seed total tocopherol content of at least 100 transgenic Brassica plants.

8. The transgenic seed of claim 1 , wherein the increased n-3 VLC-PUFAs content comprises DHA and wherein the increased total tocopherol content and the increased DHA content in the transgenic seed are positively correlated.

9. A transgenic seed of a transgenic Brassica plant, the transgenic seed comprising total tocopherol and n-3 very long chain polyunsaturated fatty acids (VLC-PUFAs), wherein the transgenic seed has increased total tocopherol content and increased n-3 VLC-PUFAs content as compared to a seed from a wild-type control Brassica plant of the same species and variety, wherein the increased n-3 VLC-PUFAs content comprises DHA; and wherein the increased total tocopherol content and the increased DHA content in the transgenic seed are positively correlated; wherein the transgenic seed comprises transgenes which comprise at least one recombinant polynucleotide encoding a delta-12-desaturase from Phythophthora species, at least one recombinant polynucleotide encoding a delta-6-desaturase from Ostreococcus species, at least one recombinant polynucleotide encoding a delta-6-elongase from Physcomitrella species or Thalassiosira species, at least one recombinant polynucleotide encoding a delta-5-desaturase from Thraustochytrium species, at least one recombinant polynucleotide encoding a delta-5-elongase from Ostreococcus species, at least one polynucleotide encoding a delta-4-desaturase from Thraustochytrium species or Pavlova species, and at least one recombinant polynucleotide encoding an omega-3-desaturase from Phythophthora species or Pythium species; and wherein expression of said transgenes in said transgenic seed results in said increase in n-3 VLC-PUFAs content and total tocopherol content in said transgenic seed.

10. The transgenic seed of claim 9 , wherein the total tocopherol content of the transgenic seed is more than 35 mg/100 g transgenic seed; and wherein the content of n-3 VLC-PUFAs in the transgenic seed is increased by at least 5%, at least 10%, at least 15%, at least 20%, or at least 30% by transgenic seed weight as compared to seed from a wild-type Brassica control plant of the same species and variety.

11. The transgenic seed of claim 9 , wherein the total tocopherol content of the transgenic seed is from about 36 mg/100 g transgenic seed to about 44 mg/100 g transgenic seed.

12. The transgenic seed of claim 11 , wherein the content of n-3 VLC-PUFAs in the transgenic seed is increased by 5% to 30% by transgenic seed weight as compared to a seed from a wild-type Brassica control plant of the same species and variety.

13. The transgenic seed of claim 9 , wherein the total tocopherol content is measured as the numeric average of transgenic seed total tocopherol content of at least 100 transgenic Brassica plants.

Assignees

• Basf Plant Science Co Gmbh

Inventors

• Andre Carl

Classifications

• C12N15/8247 Primary

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

• A61K36/31

  Brassicaceae or Cruciferae (Mustard family), e.g. broccoli, cabbage or kohlrabi · CPC title

• C12Y602/01003

  Long-chain-fatty-acid-CoA ligase (6.2.1.3) · CPC title

• C12Y114/19006

  DELTA12-fatty-acid desaturase (1.14.19.6), i.e. oleoyl-CoA DELTA12 desaturase · CPC title

• C12N9/0071

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