Nucleic acid constructs and methods for producing altered seed oil compositions
The present invention is in the field of plant genetics and provides recombinant nucleic acid molecules, constructs, and other agents associated with the coordinate manipulation of multiple genes in the fatty acid synthesis pathway. In particular, the agents of the present invention are associated with the simultaneous enhanced expression of certain genes in the fatty acid synthesis pathway and suppressed expression of certain other genes in the same pathway. Also provided are plants incorporating such agents, and in particular plants incorporating such constructs where the plants exhibit altered seed oil compositions.
1. A soybean seed exhibiting a seed oil fatty acid composition comprising an oleic acid content of about 42% to 85% by weight of the total fatty acids and a saturated fatty acid content of less than 8% by weight of the total fatty acids, wherein said seed further comprises a genome comprising
i) a nucleic acid sequence at least 95% identical to a soybean FAD2-1A intron that is between about 20 and about 420 nucleotides in length and that is capable, when expressed in a host cell, of suppressing the expression of endogenous soybean FAD2-1; and
ii) a combination of two nucleic acid sequences that are each at least 95% identical to a fragment of soybean FATB-1 sequence that is at least 25 contiguous nucleotides and selected from the group consisting of
a) soybean FATB-1 3′UTR;
b) soybean FATB-1 5′UTR; and
c) soybean FATB transit peptide encoding sequence,
wherein said combination of two sequences is capable, when expressed in a host cell, of suppressing the expression of endogenous soybean FATB-1 and said combination of two sequences is between about 40 to about 450 nucleotides in length.
2. The soybean seed of claim 1 , wherein said nucleic acid sequence at least 95% identical to a soybean FAD2-1A intron and said combination of two nucleic acid sequences are each transcribed in sense and antisense-orientations, resulting in RNA which is at least in part double-stranded.
3. The soybean seed of claim 1 , wherein said nucleic acid sequence and combination of two nucleic acid sequences that suppress the expression of endogenous soybean FAD2-1 and FATB-1 are assembled as a functional transcription unit after insertion into a plant chromosome.
4. A soybean seed exhibiting a seed oil fatty acid composition comprising an oleic acid content of about 42% to 85% by weight of the total fatty acids and a saturated fatty acid content of less than 8% by weight of the total fatty acids, wherein said seed further comprises a genome with a nucleic acid sequence comprising a fragment at least 95% identical to a soybean FAD2-1A intron that is between about 20 and about 420 nucleotides in length and a soybean FATB-1 sequence that is between about 40 to about 450 contiguous nucleotides in length, wherein said FATB-1 sequence comprises a UTR fragment.
5. The soybean seed of claim 4 , wherein said fragment at least 95% identical to a soybean FAD2-1A intron and said soybean FATB-1 sequence are each transcribed in sense- and antisense-orientations, resulting in RNA which is at least in part double-stranded.
6. The soybean seed of claim 4 , wherein said nucleic acid sequence is assembled as a functional transcription unit after insertion into a plant chromosome.
7. A soybean seed exhibiting a seed oil fatty acid composition comprising an oleic acid content of about 42% to 85% by weight of the total fatty acids and a saturated fatty acid content of less than 8% by weight of the total fatty acids, wherein said seed further comprises a genome with a nucleic acid sequence that suppresses the expression of endogenous soybean FAD2-1A, comprising a fragment at least 95% identical to a soybean FAD2-1A intron that is between about 20 and about 420 nucleotides in length; and said genome further comprises a nucleic acid sequence that increases the expression of both beta-ketoacyl-ACP synthase IV and delta-9 desaturase.
8. The soybean seed of claim 1 , wherein said oleic acid content is about 50% to about 80% by weight of the total fatty acids.
9. The soybean seed of claim 1 , wherein said oleic acid content is about 46% to about 75% by weight of the total fatty acids.
10. The soybean seed of claim 7 , wherein said oleic acid content is about 50% to about 80% by weight of the total fatty acids.
11. The soybean seed of claim 7 , wherein said oleic acid content is about 46% to about 75% by weight of the total fatty acids.
12. The soybean seed of claim 4 , wherein said oleic acid content is about 50% to about 80% by weight of the total fatty acids.
13. The soybean seed of claim 4 , wherein said oleic acid content is about 46% to about 75% by weight of the total fatty acids.
14. The soybean seed of claim 1 , wherein said nucleic acid sequence at least 95% identical to a soybean FAD2-1A intron is between about 100 to about 320 nucleotides in length and said combination of two nucleic acid sequences comprises a fragment of a FATB-1 5′UTR.
15. The soybean seed of claim 1 , wherein said nucleic acid sequence at least 95% identical to a soybean FAD2-1A intron is selected from the group consisting of about 30 to about 420, between about 40 to about 320, between about 50 to about 200, between about 50 to about 400, between about 50 to about 420, between about 60 to about 320, about 70 to about 220, between about 100 to about 200, between about 150 to about 200, between about 150 to about 220, between about 150 to about 400, between about 200 to about 300, and between about 300 to about 400 nucleotides in length.
16. The soybean seed of claim 4 , wherein said nucleic acid sequence at least 95% identical to a soybean FAD2-1A intron is between about 100 to about 320 nucleotides in length.
17. The soybean seed of claim 4 , wherein said nucleic acid sequence at least 95% identical to a soybean FAD2-1A intron is selected from the group consisting of about 30 to about 420, between about 40 to about 320, between about 50 to about 200, between about 50 to about 400, between about 50 to about 420, between about 60 to about 320, about 70 to about 220, between about 100 to about 200, between about 150 to about 200, between about 150 to about 220, between about 150 to about 400, between about 200 to about 300, and between about 300 to about 400 nucleotides in length.
18. The soybean seed of claim 7 , wherein said nucleic acid sequence at least 95% identical to a soybean FAD2-1A intron is between about 100 to about 320 nucleotides in length.
19. The soybean seed of claim 7 , wherein the length of said nucleic acid sequence at least 95% identical to a soybean FAD2-1A intron is selected from the group consisting of about 30 to about 420, between about 40 to about 320, between about 50 to about 200, between about 50 to about 400, between about 50 to about 420, between about 60 to about 320, about 70 to about 220, between about 100 to about 200, between about 150 to about 200, between about 150 to about 220, between about 150 to about 400, between about 200 to about 300, and between about 300 to about 400 nucleotides.
20. The soybean seed of claim 1 , wherein said nucleic acid sequence comprises a fragment 100% identical to a soybean FAD2-1A intron that is between about 20 and about 420 nucleotides in length and said combination of two sequences is 100% identical to a soybean FATB-1 and is between about 40 to about 450 nucleotides in length; and said genome further comprises a nucleic acid sequence that increases the expression of one or both of beta-ketoacyl-ACP synthase IV and delta-9 desaturase.
21. The soybean seed of claim 4 , wherein said nucleic acid sequence comprises a fragment 100% identical to a soybean FAD2-1A intron that is between about 20 and about 420 nucleotides in length and a soybean FATB-1 sequence that is between about 40 to about 450 contiguous nucleotides in length, wherein said FATB-1 sequence comprises a UTR fragment.
22. The soybean seed of claim 7 , wherein said nucleic acid sequence that suppresses the expression of endogenous soybean FAD2-1A comprises a fragment 100% identical to a soybean FAD2-A intron that is between about 20 and about 420 nucleotides in length.
23. A soybean seed exhibiting a seed oil fatty acid composition comprising an oleic acid content of about 42% to 85% by weight of the total fatty acids and a saturated fatty acid content of less than 8% by weight of the total fatty acids, wherein said seed further comprises a genome comprising
i) a nucleic acid sequence at least 90% identical to a soybean FAD2-1 A intron that is between about 20 and about 420 nucleotides in length and that is capable, when expressed in a host cell, of suppressing the expression of endogenous soybean FAD2-1; and
ii) a combination of two nucleic acid sequences that are each at least 90% identical to a fragment of soybean FATB-1 sequence that is at least 25 contiguous nucleotides and selected from the group consisting of
a) soybean FATB-1 3′UTR;
b) soybean FATB-1 5′UTR; and
c) soybean FATB transit peptide encoding sequence,
wherein said combination of two sequences is capable, when expressed in a host cell, of suppressing the expression of endogenous soybean FATB-1 and said combination of two sequences is between about 40 to about 450 nucleotides in length.
24. The soybean seed of claim 1 , wherein said combination of two sequences is said soybean FATB transit peptide encoding sequence and about 45 contiguous nucleotides of said soybean FATB-1 5′UTR.
25. The soybean seed of claim 1 , wherein said combination of two sequences is said soybean FATB transit peptide encoding sequence and 42 contiguous nucleotides of said soybean FATB-1 5′UTR.