Soybean Seed and Oil Compositions and Methods of Making Same

Abstract

Methods for obtaining soybean plants that produce seed with low linolenic acid levels and moderately increased oleic levels are disclosed. Also disclosed are methods for producing seed with low linolenic acid levels, moderately increased oleic levels and low saturated fatty acid levels. These methods entail the combination of transgenes that provide moderate oleic acid levels with soybean germplasm that contains mutations in soybean genes that confer low linolenic acid phenotypes. These methods also entail the combination of transgenes that provide both moderate oleic acid levels and low saturated fat levels with soybean germplasm that contains mutations in soybean genes that confer low linolenic acid phenotypes. Soybean plants and seeds produced by these methods are also disclosed.

1 .- 60 . (canceled) 
     
     
         61 . A method of producing soybean seed comprising a linolenic acid content of 0.5% to 6% of total seed fatty acids by weight and an oleic acid content of 55% to 80% of total seed fatty acids by weight, comprising the steps of:
 a) growing one or more soybean plants that comprise a transgene that decreases the expression of an endogenous soybean FAD2-1 gene and at least one loss-of-function mutation in an endogenous soybean FAD3 gene, wherein said soybean plants yield seed having a seed fatty acid composition comprising a linolenic acid content of 0.5% to 6% of total seed fatty acids by weight and an oleic acid content of 55% to 80% of total seed fatty acids by weight, and,   b) harvesting seed from said plant.   
     
     
         62 . The method of  claim 61 , wherein said soybean plants of step (a) comprise at least two loss of function mutations in at least two endogenous soybean FAD3 genes. 
     
     
         63 . The method of  claim 62 , wherein said endogenous soybean FAD3 genes are FAD3-1B and FAD3-1C. 
     
     
         64 . The method of  claim 62 , wherein said soybean plants yield seed comprising a linolenic acid content of 1 to 4% of total seed fatty acids by weight and an oleic acid content of 55% to 80% of total seed fatty acids by weight. 
     
     
         65 . The method of  claim 61 , wherein at least one of said loss-of-function mutations comprises a deletion in the FAD3-1C gene of SEQ ID NO:62. 
     
     
         66 . The method of  claim 61 , wherein at least one of said loss-of-function mutations in said soybean FAD3-1B gene comprises a substitution of a thymine residue for a cytosine residue at a position in the FAD3-1B gene sequence corresponding to nucleotide 2021 of SEQ ID NO:61. 
     
     
         67 . A soybean seed, said seed having a fatty acid composition comprising a linolenic acid content of 0.5% to 6% of total seed fatty acids by weight and an oleic acid content of 55% to 80% of total seed fatty acids by weight, a transgene that decreases the expression of an endogenous soybean FAD2-1 gene, and at least one loss-of-function mutation in an endogenous soybean FAD3 gene. 
     
     
         68 . The soybean seed of  claim 67 , said seed having a fatty acid composition comprising a linolenic acid content of 1% to 3% of total seed fatty acids by weight and an oleic acid content of 55% to 80% of total seed fatty acids by weight. 
     
     
         69 . The soybean seed of  claim 67 , wherein said plant comprises at least two loss-of-function mutations in at least two endogenous soybean FAD3 genes. 
     
     
         70 . The soybean seed of  claim 69 , wherein said endogenous soybean FAD3 genes are FAD3-1B and FAD3-1C. 
     
     
         71 . The soybean seed of  claim 67 , wherein said loss-of-function mutation comprises a deletion in the FAD3-1C gene of SEQ ID NO:62. 
     
     
         72 . The soybean seed of  claim 67 , wherein said loss-of-function mutation in said soybean FAD3-1B gene comprises a substitution of a thymine residue for a cytosine residue at a position in the FAD3-1B gene sequence corresponding to nucleotide 2021 of SEQ ID NO:61. 
     
     
         73 . A method of producing soybean seed comprising a linolenic acid content of about 0.5% to 6% of total seed fatty acids by weight, a saturated fatty acid content of about 2% to 8% by weight and an oleic acid content of 55% to 80% of total seed fatty acids by weight, comprising the steps of:
 a) growing one or more soybean plants that comprise at least one transgene that decreases the expression of both an endogenous soybean FAD2-1 and an endogenous FATB gene, and at least one loss-of-function mutation in an endogenous soybean FAD3 gene, wherein said soybean plants yield seed having a seed fatty acid composition comprising a linolenic acid content of about 0.5% to 6% of total seed fatty acids by weight, a saturated fatty acid content of about 2% to 8% by weight, and an oleic acid content of 55% to 80% of total seed fatty acids by weight; and,   b) harvesting seed from said plant.   
     
     
         74 . The method of  claim 73 , wherein said soybean plants comprise at least two loss of function mutations in at least two endogenous soybean FAD3 genes. 
     
     
         75 . The method of  claim 74 , wherein said endogenous soybean FAD3 genes are FAD3-1B and FAD3-1C. 
     
     
         76 . The method of  claim 73 , wherein said soybean plants yield seed comprising a linolenic acid content of about 1% to 3% of total seed fatty acids by weight, a saturated fatty acid content of about 2% to 8% by weight and an oleic acid content of 55% to 80% of total seed fatty acids by weight. 
     
     
         77 . The method of  claim 73 , wherein at least one of said loss-of-function mutations comprises a deletion in the FAD3-1C gene of SEQ ID NO:62. 
     
     
         78 . The method of  claim 73 , wherein at least one of said loss-of-function mutations in said soybean FAD3-1B gene comprises a substitution of a thymine residue for a cytosine residue at a position in the FAD3-1B gene sequence corresponding to nucleotide 2021 of SEQ ID NO:61. 
     
     
         79 . A soybean seed, said seed having a fatty acid composition comprising a linolenic acid content of about 0.5% to 6% of total seed fatty acids by weight, a saturated fatty acid content of about 2% to 8% by weight and an oleic acid content of 55% to 80% of total seed fatty acids by weight, at least one transgene that decreases the expression of both an endogenous soybean FAD2-1 and an endogenous FATB gene, and at least one loss-of-function mutation in an endogenous soybean FAD3 gene. 
     
     
         80 . The seed of  claim 79 , said seed having a fatty acid composition comprising a linolenic acid content of about 1% to 3% of total seed fatty acids by weight, a saturated fatty acid content of about 2% to 8% by weight, and an oleic acid content of 55% to 80% of total seed fatty acids by weight. 
     
     
         81 . The seed of  claim 79 , wherein said seed comprises at least two loss-of-function mutations in at least two endogenous soybean FAD3 genes. 
     
     
         82 . The seed of  claim 79 , wherein said endogenous soybean FAD3 genes are FAD3-1B and FAD3-1C. 
     
     
         83 . The seed of  claim 79 , wherein at least one of said loss-of-function mutations comprises a deletion in the FAD3-1C gene of SEQ ID NO:62. 
     
     
         84 . The seed of  claim 79 , wherein at least one of said loss-of-function mutations in said soybean FAD3-1B gene comprises a substitution of a thymine residue for a cytosine residue at a position in the FAD3-1B gene sequence corresponding to nucleotide 2021 of SEQ ID NO:61.