Plant seeds with altered storage compound levels, related constructs and methods involving genes encoding oxidoreductase motif polypeptides

Abstract

This invention is in the field of plant molecular biology. More specifically, this invention pertains to isolated nucleic acid fragments encoding ORM proteins in plants and seeds and the use of such fragments to modulate expression of a gene encoding ORM protein activity in a transformed host cell.

1 . A transgenic plant comprising a recombinant DNA construct comprising a polynucleotide operably linked to at least one regulatory element, wherein said polynucleotide encodes a polypeptide having an amino acid sequence of at least 70% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NO: 26, 28, 30, 32, 40, 42, 44, 46, 48, 64, 65, 66, 67, 69, 70, 102, 104, 105, 106, 108, 110, 113, 116, or 117 and wherein seed obtained from said transgenic plant has an altered i.e. increased or decreased oil, protein, starch and/or soluble carbohydrate content when compared to a control plant not comprising said recombinant DNA construct. 
     
     
         2 . A transgenic seed obtained from the transgenic plant of  claim 1  comprising a recombinant DNA construct comprising a polynucleotide operably linked to at least one regulatory element, wherein said polynucleotide encodes a polypeptide having an amino acid sequence of at least 70% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NO: 26, 28, 30, 32, 40, 42, 44, 46, 48, 64, 65, 66, 67, 69, 70, 102, 104, 105, 106, 108, 110, 113, 116, or 117 and wherein said transgenic seed has an altered oil, protein, starch and/or soluble carbohydrate content when compared to a seed from a control plant not comprising said recombinant DNA construct. 
     
     
         3 . A transgenic seed comprising:
 a recombinant DNA construct comprising:
 (a) a polynucleotide operably linked to at least one regulatory element, wherein said polynucleotide encodes a polypeptide having an amino acid sequence of at least 70% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NO: 26, 28, 30, 32, 40, 42, 44, 46, 48, 64, 65, 66, 67, 69, 70, 102, 104, 105, 106, 108, 110, 113, 116, or 117 or 
 (b) a suppression DNA construct comprising at least one regulatory element operably linked to: 
 (i) all or part of: (A) a nucleic acid sequence encoding a polypeptide having an amino acid sequence of at least 70% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NO: 26, 28, 30, 32, 40, 42, 44, 46, 48, 64, 65, 66, 67, 69, 70, 102, 104, 105, 106, 108, 110, 113, 116, or 117, or (B) a full complement of the nucleic acid sequence of (b)(i)(A); or 
 (ii) a region derived from all or part of a sense strand or antisense strand of a target gene of interest, said region having a nucleic acid sequence of at least 70% sequence identity, based on the Clustal V method of alignment, when compared to said all or part of a sense strand or antisense strand from which said region is derived, and wherein said target gene of interest encodes a ORM protein, and wherein said plant has an altered, increased or decreased oil, protein, starch and/or soluble carbohydrate content when compared to a control plant not comprising said recombinant DNA construct. 
   
     
     
         4 . The transgenic seed of  claim 1 , wherein the oil content in increased by at least 2% when compared to the oil content of a non-transgenic seed. 
     
     
         5 . A transgenic seed comprising a recombinant DNA construct comprising:
 (a) all or part of the nucleotide sequence set forth in SEQ ID NO: 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 48, 68, 101, 103, 107, 109, 111, or 114;   or (b) the full-length complement of (a):
 wherein (a) or (b) is of sufficient length to inhibit expression of endogenous ORM protein activity in a transgenic plant and further wherein said seed has an increase in oil content of at least 2%, on a dry-weight basis, as compared to seed obtained from a non-transgenic plant. 
   
     
     
         6 . A method for producing a transgenic plant, the method comprising:
 (a) transforming a plant cell with a recombinant DNA construct comprising a polynucleotide operably linked to at least one regulatory sequence, wherein the polynucleotide encodes a polypeptide having an amino acid sequence of at least 70% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NO: 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 64, 65, 66, 67, 69, 70, 102, 104, 105, 106, 108, 110, 113, 116, or 117; and regenerating a plant from the transformed plant cell.   
     
     
         7 . A method for producing transgenic seeds, the method comprising:
 (a) transforming a plant cell with a recombinant DNA construct comprising a polynucleotide operably linked to at least one regulatory sequence, wherein the polynucleotide encodes a polypeptide having an amino acid sequence of at least 70% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NO: 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 64, 65, 66, 67, 69, 70, 102, 104, 105, 106, 108, 110, 113, 116, or 117; and   (b) regenerating a transgenic plant from the transformed plant cell of (a); and   (c) selecting a transgenic plant that produces a transgenic seed having an altered oil, protein, starch and/or soluble carbohydrate content, as compared to a transgenic seed obtained from a non-transgenic plant.   
     
     
         8 . A method for producing transgenic seeds, the method comprising:
 (a) transforming a plant cell with a recombinant DNA construct comprising a polynucleotide operably linked to at least one regulatory sequence, wherein the polynucleotide encodes a polypeptide having an amino acid sequence of at least 70% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NO: 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 64, 65, 66, 67, 69, 70, 102, 104, 105, 106, 108, 110, 113, 116, or 117; and   (b) regenerating a transgenic plant from the transformed plant cell of (a); and   (c) selecting a transgenic plant that produces a transgenic seed having an increased starch content of at least 0.5% as compared to a transgenic seed obtained from a non-transgenic plant.   
     
     
         9 . A method for producing transgenic seed, the method comprising:
 (a) transforming a plant cell with a recombinant DNA construct comprising:
 (i) all or part of the nucleotide sequence set forth in SEQ ID NO: 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 48, 68, 101, 103, 107, 109, 111, or 114; or 
 (ii) the full-length complement of (i); 
   
       wherein (i) or (ii) is of sufficient length to inhibit expression of endogenous ORM protein activity in a transgenic plant;
 (b) regenerating a transgenic plant from the transformed plant cell of (a); and 
 (c) selecting a transgenic plant that produces a transgenic seed having an altered oil, protein, starch and/or soluble carbohydrate content, as compared to a transgenic seed obtained from a non-transgenic plant. 
 
     
     
         10 . A method for producing transgenic seed, the method comprising:
 (a) transforming a plant cell with a recombinant DNA construct comprising:
 (i) all or part of the nucleotide sequence set forth in SEQ ID NO: 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 48, 68, 101, 103, 107, 109, 111, or 114; or 
 (ii) the full-length complement of (i); 
   
       wherein (i) or (ii) is of sufficient length to inhibit expression of endogenous ORM protein activity in a transgenic plant;
 (b) regenerating a transgenic plant from the transformed plant cell of (a); and 
 (c) selecting a transgenic plant that produces a transgenic seed having an increase in oil content of at least 2%, on a dry-weight basis, as compared to a transgenic seed obtained from a non-transgenic plant. 
 
     
     
         11 . The transgenic seed of any one of  claim 1 ,  2 ,  3 ,  4 , or  5 , wherein the transgenic seed is obtained from a monocot or dicot plant. 
     
     
         12 . The transgenic seed of any one of  claim 1 ,  2 ,  3 ,  4 , or  5 , wherein the transgenic seed is obtained from a maize or soybean plant. 
     
     
         13 . The transgenic seed of any one of  claim 1 ,  2 ,  3 ,  4 , or  5 , wherein the at least one regulatory element is a seed-specific or seed-preferred promoter. 
     
     
         14 . The transgenic seed of any one of  claim 1 ,  2 ,  3 ,  4 , or  5 , wherein at least one regulatory element is an endosperm or embryo-specific promoter. 
     
     
         15 . The method of any one of  claim 6 ,  7 ,  8 ,  9 , or  10 , wherein the transgenic seed is obtained from a transgenic dicot plant comprising in its genome the recombinant construct. 
     
     
         16 . The method of any one of  claim 6 ,  7 ,  8 ,  9 , or  10 , wherein the dicot plant is soybean. 
     
     
         17 . Transgenic seed obtained by the method of any one of  claims 6 ,  7 ,  8 ,  9 , or  10 . 
     
     
         18 . A product and/or by-product obtained from the transgenic seed of claim of any one of  claim 6 ,  7 ,  8 ,  9 , or  10 . 
     
     
         19 . The transgenic seed obtained by the method of any one of  claim 6 ,  7 ,  8 ,  9 ,  10  or  11 , wherein the transgenic seed is obtained from a monocot or dicot plant. 
     
     
         20 . A product and/or by-product from transgenic seed of  claim 2  wherein the plant is maize or soybean. 
     
     
         21 . A product and/or by-product from the transgenic seed of  claim 3 , wherein the plant is maize or soybean. 
     
     
         22 . A product and/or by-product from the transgenic seed of  claim 4 , wherein the plant is maize or soybean. 
     
     
         23 . A product and/or by-product from the transgenic seed of  claim 5 , wherein the plant is maize or soybean. 
     
     
         24 . An isolated polynucleotide comprising:
 (a) a nucleotide sequence encoding a polypeptide required for altering i.e. increasing or decreasing oil, protein, starch and/or soluble carbohydrate content in a plant, wherein, based on the Clustal V method of alignment with pairwise alignment default parameters of KTUPLE=1, GAP PENALTY=3, WINDOW=5 and DIAGONALS SAVED=5, the polypeptide has an amino acid sequence of at least 70% sequence identity when compared to SEQ ID NO:32; 102, 104; 113, or 116; or   (b) the full complement of the nucleotide sequence of (a).   
     
     
         25 . The polynucleotide of  claim 24 , wherein the amino acid sequence of the polypeptide comprises SEQ ID NO: 32; 102, 104; 113, or 116. 
     
     
         26 . The polynucleotide of  claim 24 , wherein the nucleotide sequence comprises SEQ ID NO:31, 101, 103, 112, or 115. 
     
     
         27 . A plant or seed comprising a recombinant DNA construct, wherein the recombinant DNA construct comprises the polynucleotide of any one of  claims 24  to  26  operably linked to at least one regulatory sequence.