Compositions isolated from forage grasses and methods for their use

Abstract

Isolated polynucleotides encoding polypeptides active in the fructan, cellulose, starch and/or tannin biosynthetic pathways are provided, together with expression vectors and host cells comprising such isolated polynucleotides. Methods for the use of such polynucleotides and polypeptides are also provided.

We claim:  
     
         1 . An isolated polynucleotide comprising a sequence selected from the group consisting of: SEQ ID NO: 1-44.  
     
     
         2 . An isolated polynucleotide comprising a sequence selected from the group consisting of: 
 (a) complements of SEQ ID NO: 1-44;    (b) reverse complements of SEQ ID NO: 1-44;    (c) reverse sequences of SEQ ID NO: 1-44;    (d) sequences that are 100-mers of a sequence of SEQ ID NO: 1-44;    (e) sequences that are 40-mers of a sequence of SEQ ID NO: 1-44; and    (f) sequences that are 20-mers of a sequence of SEQ ID NO: 1-44.    
     
     
         3 . An isolated polynucleotide comprising a sequence selected from the group consisting of: 
 (a) sequences having at least 75% identity to a sequence of SEQ ID NO: 1-44;    (b) sequences having at least 90% identity to a sequence of SEQ ID NO: 1-44;    (c) sequences having at least 95% identity to a sequence of SEQ ID NO: 1-44;    (d) sequences having at least 98% identity to a sequence of SEQ ID NO: 1-44; and    (e) sequences that hybridize to a sequence of SEQ ID NO: 1-44 under stringent hybridization conditions,    wherein the polynucleotide encodes a polypeptide having substantially the same functional activity as a polypeptide encoded by a polynucleotide of SEQ ID NO: 1-44.    
     
     
         4 . An isolated polypeptide encoded by a polynucleotide of any one of claims  1 - 3 .  
     
     
         5 . An isolated polypeptide comprising an amino acid sequence selected from the group consisting of: sequences recited in SEQ ID NO: 45-88.  
     
     
         6 . An isolated polypeptide comprising an amino acid sequence selected from the group consisting of: 
 (a) sequences having at least 75% identity to a sequence of SEQ ID NO: 45-88;    (b) sequences having at least 90% identity to a sequence of SEQ ID NO: 45-88; and    (c) sequences having at least 95% identity to a sequence of SEQ ID NO: 45-88,    wherein the polypeptide has substantially the same functional properties as a polypeptide of SEQ ID NO: 45-88.    
     
     
         7 . An isolated polynucleotide that encodes a polypeptide of any one of claims  5  and  6 .  
     
     
         8 . An isolated oligonucleotide probe or primer comprising at least 10 contiguous residues complementary to 10 contiguous residues of a nucleotide sequence recited in any one of claims  1 - 3 .  
     
     
         9 . A kit comprising a plurality of oligonucleotide probes or primers of  claim 8 .  
     
     
         10 . A genetic construct comprising a polynucleotide of any one of claims  1 - 3 .  
     
     
         11 . A transgenic cell comprising a genetic construct according to  claim 10 .  
     
     
         12 . A genetic construct comprising, in the 5′-3′ direction: 
 (a) a gene promoter sequence;  
 (b) a polynucleotide sequence comprising at least one of the following: (1) a polynucleotide coding for at least a functional portion of a polypeptide of any one of claims  5  and  6 ; and (2) a polynucleotide comprising a non-coding region of a polynucleotide of any one of claims  1 - 3 ; and  
 (c) a gene termination sequence.  
 
     
     
         13 . The genetic construct of  claim 12 , wherein the polynucleotide sequence is in a sense orientation.  
     
     
         14 . The genetic construct of  claim 12 , wherein the polynucleotide is in an anti-sense orientation.  
     
     
         15 . A transgenic plant cell comprising a genetic construct of  claim 12 .  
     
     
         16 . A plant comprising a transgenic plant cell according to  claim 12 , or fruit or seeds or progeny thereof.  
     
     
         17 . The plant of  claim 16 , wherein the plant is selected from the group consisting of  Festuca arundinacea  and  Lolium perenne  species.  
     
     
         18 . A method for modulating at least one of the fructan composition, cellulose composition, starch composition and tannin composition of a plant, comprising modulating the activity of a polypeptide of any one of claims  5  and  6  in the plant.  
     
     
         19 . A method for modulating at least one of the fructan composition, cellulose composition, starch composition and tannin composition of a plant, comprising modulating the activity of a polynucleotide of any one of claims  1 - 3  in the plant.  
     
     
         20 . The method of  claim 19 , comprising stably incorporating into the genome of the plant a polynucleotide of any one of claims  1 - 3 .  
     
     
         21 . The method of  claim 19 , comprising stably incorporating into the genome of the plant a genetic construct of  claim 12 .  
     
     
         22 . A method for producing a plant having at least one of altered fructan composition, altered cellulose composition, altered starch composition and altered tannin composition, comprising: 
 (a) transforming a plant cell with a genetic construct of  claim 12  to provide a transgenic cell; and    (b) cultivating the transgenic cell under conditions conducive to regeneration and mature plant growth.    
     
     
         23 . A method for modifying the activity of a polypeptide involved in a fructan, cellulose, starch or tannin biosynthetic pathway in a plant, comprising modulating the activity of a polynucleotide of any one of claims  1 - 3  in the plant.  
     
     
         24 . The method of  claim 23 , comprising stably incorporating into the genome of the plant a genetic construct of  claim 12 .  
     
     
         25 . A method for modifying the activity of a polypeptide involved in a fructan, cellulose, starch or tannin biosynthetic pathway in a plant, comprising introducing into cells of the plant double stranded RNA corresponding to a polynucleotide of any one of claims  1 - 3 , thereby inhibiting expression of a polypeptide encoded by the polynucleotide.