Chimeric regulatory elements for gene expression in leaf mesophyll and bundle sheath cells

Abstract

The present invention provides chimeric regulatory elements constructed from gene regulatory element polynucleotide molecules isolated from the Zea mays genes fructose 1-6 bisphosphate aldolase (FDA), pyruvate orthophosphate dikinase (PPDK), or ribulose bisphosphate carboxylase activase (RUA), useful for expressing transgenes in plants. The invention further discloses compositions, polynucleotide constructs, transformed host cells, transgenic plants and seeds comprising the chimeric gene regulatory molecules, and methods for preparing and using the same.

1 . A chimeric regulatory molecule, wherein the molecule comprises
 (a) a nucleic acid sequence comprising at least 95% identity to SEQ ID NO:6; or   (b) a fragment of at least 95 contiguous nucleotides of SEQ ID NO:6 having gene regulatory activity.   
     
     
         2 - 4 . (canceled) 
     
     
         5 . The chimeric regulatory molecule of  claim 1 , comprising SEQ ID NO:6. 
     
     
         6 - 8 . (canceled) 
     
     
         9 . A polynucleotide construct comprising a gene regulatory molecule of  claim 1 . 
     
     
         10 . The polynucleotide construct of  claim 9 , wherein said chimeric regulatory molecule comprises SEQ ID NO:6. 
     
     
         11 . (canceled) 
     
     
         12 . The polynucleotide construct of  claim 9 , wherein said regulatory molecule is operably linked to a transcribable polynucleotide molecule. 
     
     
         13 . The polynucleotide construct of  claim 12 , wherein said transcribable polynucleotide molecule is a gene of agronomic interest. 
     
     
         14 . The polynucleotide construct of  claim 13 , wherein said gene of agronomic interest is selected from the group consisting of: herbicide tolerance, insect control, modified yield, fungal disease resistance, virus resistance, nematode resistance, bacterial disease resistance, plant growth and development, starch production, modified oils production, high oil production, modified fatty acid content, high protein production, fruit ripening, enhanced animal and human nutrition, biopolymers, environmental stress resistance, pharmaceutical peptides and secretable peptides, improved processing traits, improved digestibility, enzyme production, flavor, nitrogen fixation, hybrid seed production, fiber production, and biofuel production. 
     
     
         15 . A transgenic plant cell stably transformed with the polynucleotide construct of  claim 9 . 
     
     
         16 . A transgenic plant stably transformed with the polynucleotide construct of  claim 9 . 
     
     
         17 . A seed of said transgenic plant of  claim 16 , wherein the seed comprises the polynucleotide construct. 
     
     
         18 . The transgenic plant cell of  claim 15 , wherein said plant cell is from a monocotyledonous plant. 
     
     
         19 . The transgenic plant of  claim 16 , wherein said plant is a monocotyledonous plant. 
     
     
         20 . The transgenic plant cell of  claim 15 , wherein said plant cell is from a dicotyledonous plant. 
     
     
         21 . The transgenic plant of  claim 16 , wherein said plant is a dicotyledonous plant. 
     
     
         22 . A progeny of the transgenic plant of  claim 16 , wherein said progeny has inherited said construct. 
     
     
         23 . A method of inhibiting weed growth in a field of transgenic herbicide tolerant crop plants comprising planting the transgenic plants transformed with an expression cassette comprising
 a. a regulatory element polynucleotide molecule of  claim 1 , active in a plant cell and operably linked to a polynucleotide molecule encoding an herbicide tolerance gene; and   b. applying herbicide to the field at an application rate that inhibits the growth of weeds, wherein the growth and yield of the transgenic crop plant is not substantially affected by the herbicide application,   wherein said herbicide is selected from the group consisting of: glyphosate, glufosinate, sulfonylureas, imidazolinones, bromoxynil, delapon, dicamba, cyclohezanedione, protoporphyrionogen oxidase inhibitors, and isoxasflutole herbicides.   
     
     
         24 . A method of providing pest control in a field of transgenic crop plants comprising planting the transgenic plants transformed with an expression cassette comprising a regulatory element polynucleotide molecule of  claim 1 , active in a plant cell and operably linked to a gene conferring pest resistance. 
     
     
         25 . A method of providing disease control in a field of transgenic crop plants comprising planting the transgenic plants transformed with an expression cassette comprising a regulatory element polynucleotide molecule of  claim 1 , active in a plant cell and operably linked to a gene conferring disease resistance. 
     
     
         26 . A method of providing stress tolerance to plants in a field of transgenic crop plants comprising planting the transgenic plants transformed with an expression cassette comprising a regulatory element polynucleotide molecule of  claim 1 , active in a plant cell and operably linked to a gene conferring stress tolerance. 
     
     
         27 . The method of  claim 26 , wherein said stress is further described as water deficit. 
     
     
         28 . A method of providing yield enhancement to plants in a field of transgenic crop plants comprising planting the transgenic plants transformed with an expression cassette comprising a regulatory element polynucleotide molecule of  claim 1 , active in a plant cell and operably linked to a gene conferring yield enhancement. 
     
     
         29 . (canceled) 
     
     
         30 . A method of transforming a host cell comprising:
 a. providing a chimeric regulatory molecule comprising a polynucleotide selected from the group consisting (a) a polynucleotide having at least 95% identity to SEQ ID NO:6 and (b) a fragment of at least 95 contiguous nucleotides of SEQ ID NO:6 having gene regulatory activity, wherein said molecule is operably linked to a transcribable polynucleotide molecule sequence, and   b. transforming said cell with the nucleic acid molecule.   
     
     
         31 . The method of  claim 30 , wherein the polynucleotide comprises SEQ ID NO:6. 
     
     
         32 . The method of  claim 30 , wherein the polynucleotide comprises a fragment of at least 95 contiguous nucleotides of SEQ ID NO:6 having gene regulatory activity. 
     
     
         33 . The seed of  claim 17 , wherein the chimeric regulatory molecule comprises SEQ ID NO:6. 
     
     
         34 . The seed of  claim 17 , wherein the chimeric regulatory molecule comprises a fragment of at least 95 contiguous nucleotides of SEQ ID NO:6 having gene regulatory activity. 
     
     
         35 . The chimeric regulatory molecule of  claim 1 , wherein the chimeric regulatory molecule comprises a fragment of at least 95 contiguous nucleotides of SEQ ID NO:6 having gene regulatory activity.