Phytate Polynucleotides and Methods of Use

Abstract

This invention relates to newly identified polynucleotides and polypeptides in the phytic acid biosynthetic pathway, fragments, variants and derivatives of same; methods for making the polynucleotides, polypeptides, fragments, variants, derivatives and antagonists. In particular the invention relates to polynucleotides and polypeptides of the inositol 1,3,4-trisphosphate 5/6-kinase gene family. In particular this invention relates to using the newly identified polynucleotides and polypeptides to modulate the phytic acid biosynthesis in such a way as to decrease phytate and/or increase non-phytate phosphorous, especially in corn or soy animal feedstuffs.

1 . An isolated nucleic acid comprising a member selected from the group consisting of:
 (a) a polynucleotide having at least 80% identity to SEQ ID NO:11; wherein the % sequence identity is determined over the entire length of the coding sequence by the GAP algorithm using default parameters;   (b) a polynucleotide which encodes a polypeptide of SEQ ID NO:12; and   (c) a polynucleotide which is complementary to a polynucleotide of (a) through (c) wherein the isolated nucleic acid modulates the level of ITPK.   
     
     
         2 . The isolated nucleic acid of  claim 1 , wherein the polynucleotide is from a plant. 
     
     
         3 . A vector comprising at least one nucleic acid of  claim 1 . 
     
     
         4 . An expression cassette comprising at least one nucleic acid of  claim 1  operably linked to a promoter. 
     
     
         5 . The expression cassette of  claim 4  further comprising any combination of additional polynucleotide sequences of interest. 
     
     
         6 . A host cell containing at least one nucleic acid of  claim 1 . 
     
     
         7 . The host cell of  claim 6 , wherein the host cell is a bacteria, yeast or plant cell. 
     
     
         8 . A transformed plant comprising at least one nucleic acid of  claim 1 . 
     
     
         9 . The transformed plant of  claim 8 , wherein the plant is corn, barley, soybean, sorghum, wheat, rice, alfalfa, safflower, sunflower, canola, cotton, or millet. 
     
     
         10 . A transformed seed from the transformed plant of  claim 8 . 
     
     
         11 . A method for modulating inositol 1,3,4-trisphosphate 5/6-kinase (ITPK) activity or levels in a host cell, comprising:
 (a) transforming the host cell with at least one nucleic acid of  claim 1 ; and   (b) growing the transformed host cell to modulate ITPK activity in the host cell.   
     
     
         12 . The method of  claim 11 , further comprising transforming the host cell with any combination of additional polynucleotide sequences of interest. 
     
     
         13 . The method of  claim 11 , wherein the host cell is a bacteria, yeast or plant cell. 
     
     
         14 . The method of  claim 13 , wherein the host cell is a plant cell and is from a monocot or a dicot. 
     
     
         15 . The method of  claim 13 , further comprising producing a transformed plant from the plant cell. 
     
     
         16 . A method of improving the quality of animal feed, comprising:
 (a) obtaining a transformed plant cell comprising at least one nucleic acid of  claim 1 ;   (b) generating a plant from the plant cell and selecting for a transformed genotype; and   (c) producing animal feed from the plant or plant part, wherein the animal feed has improved quality over feed from non-transformed plants.   
     
     
         17 . The method of  claim 16 , further comprising a plant cell comprising any combination of additional polynucleotide sequences of interest. 
     
     
         18 . The method of  claim 16 , wherein the plant cell is from a monocot or a dicot. 
     
     
         19 . A method of decreasing the level of phosphorous in non-ruminant animal waste comprising providing the non-ruminant animal the animal feed produced by the method of  claim 18 . 
     
     
         20 . A method of increasing the level of available phosphorous in animal feed, comprising:
 (a) obtaining a transformed plant cell comprising at least one nucleic acid of  claim 1 ;   (b) generating a plant from the plant cell and selecting for a transformed genotype; and   (c) producing animal feed from the plant, wherein the animal feed has an increased level of available phosphorous.   
     
     
         21 . The method of  claim 20  further comprising a transformed plant cell comprising any combination of additional polynucleotide sequences of interest. 
     
     
         22 . The method of  claim 20 , wherein the plant cell is from a monocot or a dicot. 
     
     
         23 . A method of creating a plant with a desired phenotype comprising:
 (a) obtaining a plant cell comprising at least one ITPK polynucleotide of  claim 1  and at least one polynucleotide of interest;   (b) generating a transformed plant with a desired phenotype.   
     
     
         24 . The method of  claim 23 , wherein the polynucleotide of interest is myo-inositol monophosphatase (IMP) or phytase. 
     
     
         25 . The method of  claim 23 , wherein the polynucleotide of interest is inositol polyphosphate kinase (IPPK) or myo-inositol 1-phosphate synthase (MI1PS). 
     
     
         26 . A transformed plant produced by the method of  claim 23 . 
     
     
         27 . The transformed plant of  claim 26 , wherein the plant is corn, barley, soybean, sorghum, wheat, rice, alfalfa, safflower, sunflower, canola, cotton, or millet. 
     
     
         28 . A transformed seed from the plant of  claim 26 . 
     
     
         29 . A method for making a genetically modified plant with increased energy availability the method comprising:
 (a) transforming a cell with an expression cassette comprising an isolated nucleic acid selected from the group consisting of:
 (i) a polynucleotide of from about 21 nucleotides to about 40 nucleotides encoding an RNA that is capable of inhibiting expression of an mRNA encoded by a polynucleotide comprising a nucleic acid of  claim 1 ; or 
 (ii) a polynucleotide encoding a transcript comprising a sense strand comprising the polynucleotide of (i), and an antisense strand comprising the complement of the polynucleotide of (i), wherein the transcript encoded by the polynucleotide is capable of forming a double stranded RNA; 
   (b) regenerating a genetically modified plant from the cell; and   (c) selecting for a genetically modified plant with increased energy availability.   
     
     
         30 . An isolated polynucleotide of from about 21 nucleotides to about 40 nucleotides encoding an RNA that is capable of inhibiting expression of an mRNA encoded by a polynucleotide comprising a nucleic acid of  claim 1 . 
     
     
         31 . An isolated polynucleotide encoding a transcript comprising a sense strand of the polynucleotide of  claim 29  and an antisense strand comprising the complement of the polynucleotide of  claim 29 , wherein the transcript encoded by the polynucleotide forms a double stranded RNA (dsRNA). 
     
     
         32 . An isolated ds RNA capable of inhibiting expression of any one of the nucleic acid molecules of  claim 1 .