Materials and methods for increasing the tocopherol content in seed oil
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.
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.