Isolated Polynucleotide Molecules Corresponding to Mutant and Wild-Type Alleles of the Maize D9 Gene and Methods of Use

Abstract

The invention provides isolated polynucleotide molecules encoding mutant and wild-type alleles of the maize D9 gene. The invention further provides methods for modifying the growth of plants by decreasing or increasing plant height, altering stem and/or root growth, and involving the use of these isolated polynucleotide molecules, isolated polypeptides, and transformed plants, seeds, and cells.

1 . An isolated polypeptide comprising an amino acid sequence selected from the group consisting of:
 (a) the amino acid sequence comprising SEQ ID NO: 2 or 5;   (b) the amino acid sequence comprising at least 93% sequence identity to SEQ ID NO: 2, wherein said polypeptide has Zm-D9 activity;   (c) the amino acid sequence comprising at least 95% sequence identity to SEQ ID NO: 5, wherein said polypeptide has MUT1 Zm-D9 activity;   (d) the amino acid sequence encoded by a nucleotide sequence that hybridizes under stringent conditions to the complement of SEQ ID NO: 1 or 3, wherein said stringent conditions comprise hybridization in 50% formamide, 1 M NaCl, 1% SDS at 37° C., and a wash in 0.1×SSC at 60° C. to 65° C.;   (e) the amino acid sequence encoded by a nucleotide sequence that hybridizes under stringent conditions to the complement of SEQ ID NO: 4 or 6, wherein said stringent conditions comprise hybridization in 50% formamide, 1 M NaCl, 1% SDS at 37° C., and a wash in 0.1×SSC at 60° C. to 65° C.;   (f) the amino acid sequence comprising at least 124 consecutive amino acids of SEQ ID NO: 2, wherein said polypeptide retains Zm-D9 activity; and   (g) the amino acid sequence comprising at least 124 consecutive amino acids of SEQ ID NO: 5, wherein said polypeptide retains MUT1 Zm-D9 activity.   
     
     
         2 . A method of increasing the level of a polypeptide in a plant comprising introducing into said plant a polynucleotide molecule comprising a nucleotide sequence selected from the group consisting of:
 (a) the nucleotide sequence comprising SEQ ID NO: 1, 3, 4 or 6;   (b) the nucleotide sequence encoding an amino acid sequence comprising SEQ ID NO: 2 or 5;   (c) the nucleotide sequence that hybridizes under stringent conditions to the complement of the nucleotide sequence of (a), wherein said stringent conditions comprise hybridization in 50% formamide, 1 M NaCl, 1% SDS at 37° C., and a wash in 0.1×SSC at 60° C. to 65° C.;   (d) the nucleotide sequence comprising at least 92% sequence identity to SEQ ID NO: 1 or 3, wherein said polynucleotide molecule encodes a polypeptide having Zm-D9 activity;   (e) the nucleotide sequence comprising at least 698 consecutive nucleotides of SEQ ID NO: 1 or 3 or a complement thereof;   (f) the nucleotide sequence encoding an amino acid sequence having at least 93% sequence identity to SEQ ID NO: 2, wherein said polynucleotide molecule encodes a polypeptide having Zm-D9 activity; and   (g) the nucleotide sequence comprising at least 96% sequence identity to SEQ ID NO: 4 or 6, wherein said polynucleotide molecule encodes a polypeptide having MUT1 Zm-D9 activity;   (h) the nucleotide sequence comprising at least 395 consecutive nucleotides of SEQ ID NO: 4 or 6 or a complement thereof; and   (i) the nucleotide sequence encoding an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 5, wherein said polynucleotide molecule encodes a polypeptide having MUT1 Zm-D9 activity.   
     
     
         3 . A method for modulating the level of a polypeptide in a plant comprising introducing into said plant a polynucleotide molecule comprising a nucleotide sequence selected from the group consisting of:
 (a) the nucleotide sequence comprising SEQ ID NO: 1, 3, 4 or 6;   (b) the nucleotide sequence encoding an amino acid sequence comprising SEQ ID NO: 2 or 5;   (c) the nucleotide sequence that hybridizes under stringent conditions to the complement of the nucleotide sequence of (a), wherein said stringent conditions comprise hybridization in 50% formamide, 1 M NaCl, 1% SDS at 37° C., and a wash in 0.1×SSC at 60° C. to 65° C.;   (d) the nucleotide sequence comprising at least 92% sequence identity to SEQ ID NO: 1 or 3, wherein said polynucleotide molecule encodes a polypeptide having Zm-D9 activity;   (e) the nucleotide sequence comprising at least 698 consecutive nucleotides of SEQ ID NO: 1 or 3 or a complement thereof;   (f) the nucleotide sequence encoding an amino acid sequence having at least 93% sequence identity to SEQ ID NO: 2, wherein said polynucleotide molecule encodes a polypeptide having Zm-D9 activity; and   (g) the nucleotide sequence comprising at least 96% sequence identity to SEQ ID NO: 4 or 6, wherein said polynucleotide molecule encodes a polypeptide having MUT1 Zm-D9 activity;   (h) the nucleotide sequence comprising at least 395 consecutive nucleotides of SEQ ID NO: 4 or 6 or a complement thereof; and   (i) the nucleotide sequence encoding an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 5, wherein said polynucleotide molecule encodes a polypeptide having MUT1 Zm-D9 activity.   
     
     
         4 . The method of  claim 2  or  3 , wherein said polynucleotide molecule is stably integrated into the genome of the plant. 
     
     
         5 . The method of any one of  claims 2 - 4 , wherein said plant is a plant cell. 
     
     
         6 . The method of any one of  claims 2 - 4 , wherein said plant is a dicot. 
     
     
         7 . The method of  claim 6 , wherein said dicot is selected from the group consisting of  Arabidopsis , soybean, sunflower, safflower, alfalfa,  Brassica , cotton, and peanut. 
     
     
         8 . The method of any one of  claims 2 - 4 , wherein said plant is a monocot. 
     
     
         9 . The method of  claim 8 , wherein said monocot is selected from the group consisting of maize, wheat, rice, sorghum, rye, millet and barley. 
     
     
         10 . The method of any one of  claim 2 , wherein said plant is a seed. 
     
     
         11 . A method for modifying the growth of a plant, said method comprising transforming an organism with a polynucleotide construct comprising a nucleotide sequence operably linked to a promoter capable of driving the expression of said nucleotide sequence in said organism, wherein said nucleotide sequence is selected from the group consisting of:
 (a) the nucleotide sequence comprising SEQ ID NO: 1, 3, 4 or 6;   (b) the nucleotide sequence encoding an amino acid sequence comprising SEQ ID NO: 2 or 5;   (c) the nucleotide sequence that hybridizes under stringent conditions to the complement of the nucleotide sequence of (a), wherein said stringent conditions comprise hybridization in 50% formamide, 1 M NaCl, 1% SDS at 37° C., and a wash in 0.1×SSC at 60° C. to 65° C.;   (d) the nucleotide sequence comprising at least 92% sequence identity to SEQ ID NO: 1 or 3, wherein said polynucleotide molecule encodes a polypeptide having Zm-D9 activity;   (e) the nucleotide sequence comprising at least 698 consecutive nucleotides of SEQ ID NO: 1 or 3 or a complement thereof;   (f) the nucleotide sequence encoding an amino acid sequence having at least 93% sequence identity to SEQ ID NO: 2, wherein said polynucleotide molecule encodes a polypeptide having Zm-D9 activity; and   (g) the nucleotide sequence comprising at least 96% sequence identity to SEQ ID NO: 4 or 6, wherein said polynucleotide molecule encodes a polypeptide having MUT1 Zm-D9 activity;   (h) the nucleotide sequence comprising at least 395 consecutive nucleotides of SEQ ID NO: 4 or 6 or a complement thereof; and   (i) the nucleotide sequence encoding an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 5, wherein said polynucleotide molecule encodes a polypeptide having MUT1 Zm-D9 activity.   
     
     
         12 . The method of  claim 11 , wherein said nucleotide sequence is operably linked to said promoter for the production of antisense transcripts. 
     
     
         13 . The method of  claim 11 , wherein the height of said plant is decreased when compared to an untransformed plant. 
     
     
         14 . The method of  claim 11 , wherein the height of said plant is increased when compared to an untransformed plant. 
     
     
         15 . The method of  claim 14 , wherein the root architecture of said plant is modified when compared to an untransformed plant. 
     
     
         16 . The method of  claim 11 , wherein said plant is a monocot. 
     
     
         17 . The method of  claim 16 , wherein said monocot is selected from the group consisting of maize, wheat, rice, sorghum, rye, millet and barley. 
     
     
         18 . The method of  claim 11 , wherein said plant is a dicot. 
     
     
         19 . The method of  claim 18 , wherein said dicot is selected from the group consisting of  Arabidopsis , soybean, sunflower, safflower, alfalfa,  Brassica , cotton, and peanut.