Regulatory Elements Associated with CBF Transcription Factors of Maize

Abstract

Compositions and methods for regulating expression of a polynucleotide of interest in a plant are provided. Compositions include novel nucleotide sequences comprising an isolated stress-induced promoter natively linked to the maize ZmCBF2 coding region. A method for expressing a polynucleotide of interest in a plant or plant cell, using a regulatory sequence described herein, is provided. The method may comprise transforming a plant cell to comprise a polynucleotide sequence of interest operably linked to one or more of the regulatory sequences of the present invention and regenerating a stably transformed plant from the transformed plant cell.

1 . An isolated nucleic acid molecule comprising a polynucleotide which has promoter activity and is selected from the group consisting of:
 a) the polynucleotide shown as positions 1-2209 of SEQ ID NO: 2;   b) a polynucleotide comprising at least 2000 contiguous nucleotides of positions 1-2209 of SEQ ID NO: 2; and   c) a polynucleotide at least 95% identical to positions 1-2209 of SEQ ID NO: 2 and retaining all regulatory elements identified at positions 64-68, 318-323, 442-447, 873-877, 1160-1164, 1179-1184, 1209-1214, 1264-1269, 1483-1488, 1598-1603, 1721-1726, 1746-1751, 1957-1962, and 2081-2086 of SEQ ID NO: 2.   
     
     
         2 . An expression cassette comprising the polynucleotide of  claim 1  operably linked to a polynucleotide of interest, wherein the polynucleotide of  claim 1  is heterologous to the polynucleotide of interest. 
     
     
         3 . A vector comprising the expression cassette of  claim 2 . 
     
     
         4 . A plant cell having stably incorporated into its genome the expression cassette of  claim 2 . 
     
     
         5 . The plant cell of  claim 4 , wherein said plant cell is from a monocot. 
     
     
         6 . The plant cell of  claim 5 , wherein said monocot is maize, barley, wheat, oat, rye, sorghum or rice. 
     
     
         7 . A plant having stably incorporated into its genome the expression cassette of  claim 2 . 
     
     
         8 . The plant of  claim 7 , wherein said plant is a monocot. 
     
     
         9 . The plant of  claim 8 , wherein said monocot is maize, barley, wheat, oat, rye, sorghum or rice. 
     
     
         10 . A transgenic seed of the plant of  claim 7 . 
     
     
         11 . The plant of  claim 7 , wherein the polynucleotide of interest encodes a gene product that confers drought tolerance, cold tolerance or salt tolerance. 
     
     
         12 . The plant of  claim 7 , wherein the polynucleotide of interest encodes a polypeptide involved in nutrient uptake, nitrogen use efficiency, root strength, root lodging resistance, soil pest management, corn rootworm resistance, carbohydrate metabolism, protein metabolism, fatty acid metabolism or phytohormone biosynthesis. 
     
     
         13 . A method for expressing a polynucleotide of interest in a plant, said method comprising introducing into the plant an expression cassette comprising a promoter and the polynucleotide of interest operably linked thereto, wherein said promoter comprises a polynucleotide selected from the group consisting of:
 a) the polynucleotide shown as positions 1-2209 of SEQ ID NO: 2;   b) a polynucleotide comprising at least 2000 contiguous nucleotides of positions 1-2209 of SEQ ID NO: 2; and   c) a polynucleotide at least 95% identical to positions 1-2209 of SEQ ID NO: 2 and retaining all regulatory elements identified at positions 64-68, 318-323, 442-447, 873-877, 1160-1164, 1179-1184, 1209-1214, 1264-1269, 1483-1488, 1598-1603, 1721-1726, 1746-1751, 1957-1962, and 2081-2086 of SEQ ID NO: 2.   and whereby the polynucleotide of interest is expressed in the plant.   
     
     
         14 . The method of  claim 13 , wherein said plant is a monocot. 
     
     
         15 . The method of  claim 14 , wherein said monocot is maize, barley, wheat, oat, rye, sorghum or rice. 
     
     
         16 . The method of  claim 13 , wherein said polynucleotide of interest encodes a gene product that confers drought tolerance, cold tolerance or salt tolerance. 
     
     
         17 . The method of  claim 13 , wherein said polynucleotide of interest encodes a polypeptide involved in nutrient uptake, nitrogen use efficiency, root strength, root lodging resistance, corn rootworm resistance, carbohydrate metabolism, protein metabolism, fatty acid metabolism or phytohormone biosynthesis. 
     
     
         18 . A method for expressing a polynucleotide of interest in a plant cell, said method comprising introducing into the plant cell an expression cassette comprising a promoter and a polynucleotide of interest operably linked thereto, wherein said promoter comprises a polynucleotide selected from the group consisting of:
 a) the polynucleotide shown as positions 1-2209 of SEQ ID NO: 2;   b) a polynucleotide comprising at least 2000 contiguous nucleotides of positions 1-2209 of SEQ ID NO: 2; and   (c) a polynucleotide at least 95% identical to positions 1-2209 of SEQ ID NO: 2 and retaining all regulatory elements identified at positions 64-68, 318-323, 442-447, 873-877, 1160-1164, 1179-1184, 1209-1214, 1264-1269, 1483-1488, 1598-1603, 1721-1726, 1746-1751, 1957-1962, and 2081-2086 of SEQ ID NO: 2.   and whereby the polynucleotide of interest is expressed in the plant cell.   
     
     
         19 . The method of  claim 18 , wherein said polynucleotide of interest encodes a gene product that confers drought tolerance, cold tolerance or salt tolerance. 
     
     
         20 . The method of  claim 18 , wherein said polynucleotide of interest encodes a polypeptide involved in nutrient uptake, nitrogen use efficiency, root strength, root lodging resistance, corn rootworm resistance, carbohydrate metabolism, protein metabolism, fatty acid metabolism, or phytohormone biosynthesis.