Modification of Lignin Biosynthesis Via Sense Suppression

Abstract

The present invention relates to the modification of lignin biosynthesis in plants, to enzymes involved in the lignin biosynthetic pathway and nucleic acids encoding such enzymes and, more particularly, to methods of modifying lignin biosynthesis via sense suppression and to related nucleic acids and constructs.

1 - 26 . (canceled) 
     
     
         27 . A substantially purified or isolated nucleic acid comprising a fragment or variant of a gene encoding a lignin biosynthetic enzyme, said nucleic acid being capable of modifying lignin biosynthesis in a plant via sense suppression;
 wherein said fragment or variant comprises a frame shift mutation relative to the gene upon which the fragment or variant is based, resulting in a loss or substantial reduction in enzymatic activity in the encoded polypeptide; and   wherein said frame shift mutation is a mutation that inserts or deletes a number of nucleotides that is not evenly divisible by three within a short distance of the ATG start codon of the gene upon which the fragment or variant is based.   
     
     
         28 . The nucleic acid according to  claim 27 , wherein said lignin biosynthetic enzyme is selected from the group consisting of 4CL, CCR, CAD, C3H, C4H, CCoAOMT, COMT, F5H and PAL. 
     
     
         29 . The nucleic acid according to  claim 27 , from a forage, turf or bioenergy grass species. 
     
     
         30 . The nucleic acid according to  claim 29 , from a  Lolium, Festuca, Cynodon, Bracharia, Paspalum, Panicum, Miscanthus, Pennisetum , or  Phalaris species.    
     
     
         31 . The nucleic acid according to  claim 27 , wherein said gene encodes 4CL and comprises a nucleotide sequence selected from the group consisting of the Seq, ID Nos: 1, 3, 5, 21, 25, 29, 33, 37, and 235-243; and wherein said nucleic acid is capable of modifying lignin biosynthesis in a plant via sense suppression of a gene encoding 4CL in said plant. 
     
     
         32 . The nucleic acid according to  claim 27  wherein said gene encodes CCR and comprises a nucleotide sequence selected from the group consisting of sequences shown in Sequence ID No: 7, 117, 121 and 244 to 251 and wherein said nucleic acid is capable of modifying lignin biosynthesis in a plant via sense suppression of a gene encoding CCR in said plant. 
     
     
         33 . The nucleic acid according to  claim 27 , wherein said gene encodes CAD and comprises a nucleotide sequence selected from the group consisting of the sequences shown in Sequence ID Nos: 9, 11, 14, 16 53, 57, 252 to 269 and 361; and wherein said nucleic acid is capable of modifying lignin biosynthesis in a plant via sense suppression of a gene encoding CAD in said plant. 
     
     
         34 . The nucleic acid according to  claim 27 , wherein said gene encodes C4H and comprises a nucleotide sequence selected from the group consisting of Sequence ID Nos: 41, 45, 49, and 270-273; and wherein said nucleic acid is capable of modifying lignin biosynthesis in a plant via sense suppression of a gene encoding C4H in said plant. 
     
     
         35 . The nucleic acid according to  claim 27 , wherein said gene encodes CCoAOMT and comprises a nucleotide sequence selected from the group consisting of Sequence ID Nos: 89, 93, 274 to 294 and 362; and wherein said nucleic acid is capable of modifying lignin biosynthesis in a plant via sense suppression of a gene encoding CCoAOMT in said plant. 
     
     
         36 . The nucleic acid according to  claim 27 , wherein said gene encodes COMT and comprises a nucleotide sequence selected from the group consisting of Sequence ID Nos: 133, 137, 141, 145, 295 to 342, 360, 362 and 362; and wherein said nucleic acid is capable of modifying lignin biosynthesis in a plant via sense suppression of a gene encoding COMT in said plant. 
     
     
         37 . The nucleic acid according to  claim 27 , wherein said gene encodes F5H and comprises a nucleotide sequence selected from the group consisting of Sequence ID Nos: 173, 177, and 343- to 346; and wherein said nucleic acid is capable of modifying lignin biosynthesis in a plant via sense suppression of a gene encoding F5H in said plant. 
     
     
         38 . The nucleic acid according to  claim 27 , wherein said gene encodes PAL and comprises a nucleotide sequence selected from the group consisting of Sequence ID Nos: 181, 185, 189, 193, 197, 201, 205, 209, 213, 217, 220, 224 and 347 to 358; and wherein said nucleic acid is capable of modifying lignin biosynthesis in a plant via sense suppression of a gene encoding PAL in said plant. 
     
     
         39 . The nucleic acid according to  claim 27 , wherein said gene encodes C3H and comprises a nucleotide sequence shown in SEQ ID No: 359; and wherein said nucleic acid is capable of modifying lignin biosynthesis in a plant via sense suppression of a gene encoding C3H in said plant. 
     
     
         40 . The nucleic acid according to  claim 27 , comprising a nucleotide sequence selected from the group consisting of SEQ ID Nos: 23, 27, 31, 35, 39, 43, 47, 51, 55, 59, 63, 67, 71, 75, 79, 83, 87, 91, 95, 99, 103, 107, 111, 115, 119, 123, 127, 131, 135, 139, 143, 147, 151, 155, 159, 163, 167, 171, 175, 179, 183, 187, 191, 195, 199, 203, 207, 211, 215, 218, 222, 225, 229 and 233. 
     
     
         41 . The nucleic acid according to  claim 27  encoding a polypeptide comprising a sequence selected from the group consisting of SEQ ID Nos: 24, 28, 32, 36, 40, 44, 48, 56, 60, 64, 68. 72, 76, 80, 84, 88, 92, 96, 100, 104, 108, 112, 116, 120, 124, 128, 132, 136, 140, 144, 148, 152, 156, 160, 164, 168, 172, 176, 180, 184, 188, 192, 196, 200, 204, 208, 212, 216, 219, 223, 226, 230 and 234. 
     
     
         42 . A genetic construct or vector comprising the nucleic acid according to  claim 27 . 
     
     
         43 . A transformed plant, plant cell, plant seed or other plant part or transformed plant biomass comprising the nucleic acid according to  claim 27 . 
     
     
         44 . A method of modifying lignin biosynthesis in a plant, said method including introducing into said plant in a sense orientation an effective amount of a nucleic acid according to  claim 27 , such that expression of the corresponding gene is suppressed. 
     
     
         45 . The method according to  claim 44 , wherein said plant is selected from the group consisting of  Lolium, Festuca, Cynodon, Bracharia, Paspalum, Panicum, Miscanthus, Pennisetum, Phalaris , and other forage and turf grasses, corn, grains, oat, sugarcane, wheat, barley,  Arabidopsis , tobacco, legumes, Alfalfa, oak,  Eucalyptus , maple,  Populus , canola, soybean, chickpea and  Pinus.    
     
     
         46 . The method according to  claim 44 , wherein said nucleic acid is part of a genetic construct or vector.