Compositions and methods for modification of fatty acids in soybean
AssigneePioneer Hi-Bred International, Inc.
InventorsJOHN D EVERARD, ANTHONY J KINNEY, ZHAN-BIN LIU
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Abstract
Methods and compositions to modify the fatty acid profiles of seeds and the oils produced therefrom include the modification of four or more alleles of fatty acid desaturases in the genome of a plant. Compositions, polynucleotide constructs, transformed and modified host cells, and plants and seeds exhibiting altered fatty characteristics are provided.
Claims28
A method of altering the fatty acid profile in the seed of a soybean plant, the method comprising introducing four or more nucleotide modifications through four or more targeted DNA breaks at four or more genomic loci of a plant, wherein
- (a) the genomic loci comprise a first polynucleotide encoding a FAD2 polypeptide and second polynucleotide encoding a FAD3 polypeptide, and
- (b) wherein the oleic acid content is increased to at least 75% by weight of the total fatty acids and the linolenic acid content is decreased to less than 3% by weight of the total fatty acids in the seed compared to the seed of a control plant not comprising the four or more introduced nucleotide modifications.
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A soybean plant comprising a first, second, third and fourth modified genomic locus, each locus comprising one or more mutations compared to a control plant and wherein the first, second, third and fourth modified genomic loci encode a polypeptide that is at least 95% identical to the amino acid sequence of SEQ ID NOS:
- 70, 72, 74 and 76, respectively, and wherein the soybean plant has an altered fatty acid profile, produces seeds comprising at least 75% oleic acid by weight of the total fatty acids and less than 3% linolenic acid by weight of the total fatty acids, and contains no heterologous or foreign DNA at the first, second, third and fourth modified genomic loci.
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A method of detecting the presence of a polynucleotide comprising SEQ ID NOs:
- 36-43, 45-52, 54, 55, 57, 58, 60, 61, 63, 64, 77, 84-91, 93-100, 102-109, 111-118 or 136-145, the polynucleotide indicating the presence of a deletion modification in a FAD2-1 or FAD3 allele, the method comprising the steps of:
- (a) contacting a DNA sample obtained from a soybean plant or part thereof with a first and a second DNA primer molecule, the first DNA primer molecule comprising at least 12 contiguous nucleotides of SEQ ID NOs:
- 36-43, 45-52, 54, 55, 57, 58, 60, 61, 63, 64, 77, 84-91, 93-100, 102-109, 111-118 or 136-145, the contiguous nucleotides comprising the junction sequence nucleotides at positions 18-19 for SEQ ID NOs:
- 43, 123, 130 and 134;
- positions 19-20 for SEQ ID NO:
- 143;
- positions 20-21 for SEQ ID NOs:
- 60-61, 63-64, 116, 120-122, 127, 131, 133 and 144;
- positions 20-22 for SEQ ID NO:
- 145;
- positions 21-22 for SEQ ID NOs:
- 78-79, 124, 125, 128, 129, 132 and 135;
- positions 22-23 for SEQ ID NOs:
- 36 and 117;
- positions 23-24 for SEQ ID NOs:
- 55, 94 and 100;
- positions 24-25 for SEQ ID NOs:
- 106, 137 and 142;
- positions 25-26 for SEQ ID NOs:
- 47 and 88;
- positions 26-27 for SEQ ID NOs:
- 46, 48, 87, 91, 98, 99, 108, 112, 114 and 141;
- positions 27-28 for SEQ ID NOs:
- 37-40, 45, 50-52, 54, 57, 58, 84, 90, 93, 97, 103, 105, 107, 111, 115, 136 and 138;
- positions 27-29 for SEQ ID NOs:
- 49 and 139;
- positions 28-29 for SEQ ID NOs:
- 41, 42, 85, 86, 89, 95, 96, 102, 104, 109, 113 and 118;
- positions 29-30 for SEQ ID NO:
- 140;
- or the reverse complement thereof, and the second DNA molecule complementary to at least 12 contiguous nucleotides of soybean genomic DNA in proximity to and upstream or downstream of the binding site of the first DNA primer molecule;
- (b) providing a nucleic acid amplification reaction condition;
- (c) performing the nucleic acid amplification reaction, thereby producing a DNA amplicon molecule indicating the presence of a deletion modification in a FAD2-1 or FAD3 allele;
- and
- (d) detecting the DNA amplicon molecule.
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Dependencies are inferred from the claim text.