High expression cereal phytase gene

Abstract

The present invention provides mutant cereal plants and mature grain thereof, characterised by enhanced levels of the enzyme phytase in the grain, and methods for inducing, detecting and selecting the mutant cereal plants. The invention further relates to animal feed comprising said grain having enhanced amounts of phytase.

1 - 15 . (canceled) 
     
     
         16 : A  Tricticum  variety capable of producing an average phytase endosperm content of greater than 4300 FTU/kg, produced by the following:
 a) obtaining a  Tricticum  plant which has been identified as having a nucleotide at the 5′ end of a polynucleotide having the nucleotide sequence ACA VGA GTC ATG CAT [SEQ ID NO:1] wherein V is a cytosine.   b) breeding said  Tricticum  plant comprising said nucleotide sequence with a second  Tricticum  variety plant which produces an average phytase content of about 1200 FTU/kg, to produce grains,   c) growing at least one  Tricticum  plant from said grains;   d) assaying said grains to confirm the presence of said nucleotide sequence in that said variety produces an average phytase endosperm content of greater than 4300 FTU/kg.   
     
     
         17 : The variety of  claim 16  wherein said  Tricticum  plant has been identified by
 a) obtaining a sample of nucleic acids from a  Tricticum  plant or portion thereof; 
 b) detecting in said sample the presence of the nucleotide t the 5′ end of a polynucleotide having the nucleotide sequence ACA VGA GTC ATG CAT [SEQ ID NO:1] wherein V is a cytosine. 
 
     
     
         18 : The plant or variety according to  claim 17 , wherein said sample of nucleic acids comprises a first polynucleotide located 5′ upstream of and operably linked to a second polynucleotide, wherein said first polynucleotide comprises the nucleotide sequence ACA VGA GTC ATG CAT (SEQ ID NO:1) [SEQ ID NO:1] wherein V is a cytosine, and wherein said second polynucleotide encodes a phytase polypeptide having myo-inositol hexakisphosphate phosphohydrolase activity. 
     
     
         19 : A population of  Tricticum  plants grown from variety of  claim 16  and having an average phytase grain content of greater than 4300 FTU/kg. 
     
     
         20 : Grain having an average phytase content of greater than 4300FTU/kg, said grain harvested from the population of  claim 19 . 
     
     
         21 : A method of producing a milled cereal composition having an average phytase content of greater than 4300FTU/kg, comprising:
 Obtaining the grain of  claim 20  and   milling said grain to produce a milled cereal composition.   
     
     
         22 : The method of  claim 21  further comprising the step of
 cleaning or conditioning the grain. 
 
     
     
         23 : The method of  claim 21  further comprising the step of
 gristing the grain. 
 
     
     
         24 : The method of  claim 21  further comprising the step of
 breaking the grain into its cereal parts cereal grain germ, bran and endosperm. 
 
     
     
         25 : The method of  claim 21  wherein said milled composition is flour. 
     
     
         26 : A milled grain composition produced by the method of  claim 21 . 
     
     
         27 : The method of  claim 21  further comprising the step a
 combining said cereal grain composition with fodder ingredients to form an animal feed. 
 
     
     
         28 : The method of  claim 21  further comprising the step of reducing the microbial population in said animal feed. 
     
     
         29 : The method of  claim 28  where said microbial population is reduced by introducing said animal feed to a using a steam pelleting machine for a sufficient time to reduce the microbial population to safe levels for animal consumption. 
     
     
         30 : The method of  claim 28  further comprising the step of pelleting said animal feed. 
     
     
         31 : An animal feed comprising the milled cereal gain with an average phytase activity of about 4300 FTU/kg produced by the method of  claim 21 . 
     
     
         32 : Flour having an average phytase activity of about 4300 FTU/kg produced by the method of  claim 21 . 
     
     
         33 : Bread dough with enhanced inorganic phosphate levels and mineral content and improved dough mixing made from the flour of  claim 32 . 
     
     
         34 : A  Tricticum  variety produced by the following:
 a) Obtaining a plant with capable of producing an average phytase endosperm content of greater than 4300 FTU/kg;   b) Cultivating said plant to produce sufficient grain for breeding;   c) Growing said grain   d) Determining the presence of a nucleotide sequence ACA VGA GTC ATG CAT (SEQ ID NO: 1), and wherein said V is a cytosine said plant producing an average phytase endosperm content of greater than 4300 FTU/kg;   e) Repeating steps c and d until a variety with an average average phytase endosperm content of greater than 4300 FTU/kg is created.   
     
     
         35 : A  Tricticum  plant wherein an ancestor of said plant is the plant of  claim 16 . 
     
     
         36 : A cultivated  Tricticum  variety having a nucleotide sequence ACA VGA GTC ATG CAT [SEQ ID NO:1] wherein V is a cytosine, capable of producing grain with an average phytase endosperm content of greater than 4300 FTU/kg.