Elevation of oil in monocot plants
Methods of making crop plants having higher oil levels in their seeds by increasing glycolytic flux through over-expression of nucleic acids encoding phosphofructokinase are provided. The invention may further comprise the over-expression of nucleic acids encoding a pyruvate kinase to alter oil content in plant seeds, and monocot cells and plants transformed with phosphofructokinase, or phosphofructokinase and pyruvate kinase transgenes.
1 . A method of producing a monocot plant having increased oil in its seed, comprising introducing into said plant a polynucleotide encoding a phosphofructokinase, operably linked to a seed-enhanced promoter whereby the oil content of the seed is increased as compared to a seed of an isogenic plant lacking the nucleic acid sequence.
2 . The method of claim 1 , wherein the polynucleotide encoding a phosphofructokinase comprises a sequence other than SEQ ID NO:9 or SEQ ID NO:13.
3 . The method of claim 1 , wherein the polynucleotide encoding a phosphofructokinase is operably linked to a polynucleotide encoding a plastid transit peptide except when said seed-enhanced promoter is an embryo-enhanced promoter.
4 . The method of claim 1 , wherein the polynucleotide comprises a nucleic acid sequence selected from the group consisting of:
(a) a nucleic acid sequence of SEQ ID NO:1 or SEQ ID NO:11; and
(b) a nucleic acid sequence that encodes the polypeptide sequence of SEQ ID NO:2, or SEQ ID NO:12.
5 . The method of claim 4 , wherein the polynucleotide comprises a nucleic acid sequence that hybridizes to the sequence of (a) or (b) or a complement thereof under high stringency conditions of about 0.2×SSC and 65° C.
6 . The method of claim 4 , wherein the plant further comprises a polynucleotide encoding a pyruvate kinase operably linked to a seed-enhanced promoter.
7 . The method of claim 6 , wherein the polynucleotide encoding a pyruvate kinase comprises a nucleic acid sequence selected from the group consisting of:
(a) a nucleic acid sequence comprising the sequence of SEQ ID NO:3; and
(b) a nucleic acid sequence that encodes the polypeptide sequence of SEQ ID NO:4.
8 . The method of claim 7 , wherein the polynucleotide comprises a nucleic acid sequence that hybridizes to the sequence of (a) or (b) or a complement thereof under high stringency conditions of about 0.2×SSC and 65° C.
9 . The method of claim 1 , wherein the plant is a monocot selected from the group consisting of corn ( Zea mays ), rice ( Oryza sativa ), barley ( Hordeum vulgare ), millet ( Panicum miliaceum ), rye ( Secale cereale ), wheat ( Triticum aestivum ), and sorghum ( Sorghum bicolor ).
10 . The method of claim 1 , wherein the promoter is selected from the group consisting of embryo-enhanced promoters, endosperm-enhanced promoters and embryo- and endosperm-enhanced promoters.
11 . A monocot plant comprising a polynucleotide encoding a phosphofructokinase, operably linked to a seed-enhanced promoter.
12 . The plant of claim 11 , wherein the polynucleotide encoding a phosphofructokinase comprises a sequence other than SEQ ID NO:9 or SEQ ID NO:13.
13 . The plant of claim 11 , wherein the polynucleotide encoding a phosphofructokinase is linked to a polynucleotide encoding a plastid transit peptide except when said seed-enhanced promoter is an embryo-enhanced promoter.
14 . A monocot plant cell comprising a polynucleotide encoding a phosphofructokinase, operably linked to a seed-enhanced promoter.
15 . A seed produced from the plant of claim 11 , comprising a polynucleotide encoding a phosphofructokinase according to claim 7 .
16 . A meal produced from the seed of claim 15 comprising a polynucleotide encoding a phosphofructokinase according to claim 11 .
17 . An animal feed composition produced from the seed of claim 15 comprising a polynucleotide encoding a phosphofructokinase according to claim 11 .
18 . A human food composition produced from the seed of claim 15 comprising a polynucleotide encoding a phosphofructokinase according to claim 11 .
19 . An animal feed composition comprising the meal of claim 16 comprising a polynucleotide encoding a phosphofructokinase according to claim 11 .
20 . A method of making a monocot plant oil comprising the steps of:
a) growing a transformed monocot plant comprising a polynucleotide encoding a phosphofructokinase operably linked to a seed-enhanced promoter, to produce seed; and
b) processing the seed to obtain the oil.
21 . The method of claim 20 , wherein the polynucleotide encoding a phosphofructokinase comprises a sequence other than SEQ ID NO:9 or SEQ ID NO:13.
22 . The method of claim 20 , wherein the polynucleotide encoding a phosphofructokinase is linked to a polynucleotide encoding a plastid transit peptide except when said seed-enhanced promoter is an embryo-enhanced promoter.