Plant genes involved in nitrate uptake and metabolism

Abstract

The present invention relates nucleic acid molecules that are modulated (e.g., upregulated) by nitrogen in corn, to proteins or polypeptides encoded by these nucleic acid molecules, and promoters of these nucleic acid molecules. The present invention relates to a nucleic acid construct having a nucleic acid molecule that is modulated by nitrogen in corn, as well as to expression systems, host cells, plants, and plant seeds having the nucleic acid construct. The present invention also relates to a method of expressing the nucleic acid molecule that is modulated by nitrogen in a plant by growing a transgenic plant or a plant grown from a transgenic seed transformed with the construct. The present invention further relates to an isolated DNA promoter that can be used to direct nitrogen-regulated expression of an isolated nucleic acid in plants.

1 . A nucleic acid construct comprising:
 a nucleic acid molecule that is modulated by nitrogen in corn;   a 5′ DNA promoter sequence; and   a 3′ terminator sequence, wherein the nucleic acid molecule, the DNA promoter sequence, and the terminator sequence are operatively coupled to permit transcription of the nucleic acid molecule.   
     
     
         2 . The nucleic acid construct according to  claim 1 , wherein the nucleic acid molecule either:
 (a) encodes a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:8, SEQ ID NO:11, SEQ ID NO:14, SEQ ID NO:17, SEQ ID NO:20, SEQ ID NO:23, SEQ ID NO:25, SEQ ID NO:28, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:35, SEQ ID NO:37, SEQ ID NO:40, SEQ ID NO:43, and SEQ ID NO:45; or   (b) comprises a nucleotide sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:10, SEQ ID NO:13, SEQ ID NO:16, SEQ ID NO:19, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:27, SEQ ID NO:30, SEQ ID NO:34, SEQ ID NO:36, SEQ ID NO:39, SEQ ID NO:42, and SEQ ID NO:44; or   (c) comprises a coding portion of a nucleotide sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:10, SEQ ID NO:13, SEQ ID NO:16, SEQ ID NO:19, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:27, SEQ ID NO:30, SEQ ID NO:34, SEQ ID NO:36, SEQ ID NO:39, SEQ ID NO:42, and SEQ ID NO:44.   
     
     
         3 . The nucleic acid construct according to  claim 1 , wherein the DNA promoter sequence is a constitutive plant promoter. 
     
     
         4 . The nucleic acid construct according to  claim 1 , wherein the DNA promoter sequence is an inducible plant promoter. 
     
     
         5 . The nucleic acid construct according to  claim 4 , wherein the inducible plant promoter is a nitrogen inducible plant promoter. 
     
     
         6 . The nucleic acid construct according to  claim 5 , wherein the nitrogen inducible plant promoter comprises a nucleotide sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:6, SEQ ID NO:9, SEQ ID NO:12, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:21, SEQ ID NO:26, SEQ ID NO:29, SEQ ID NO:33, SEQ ID NO:38, SEQ ID NO:41, and SEQ ID NO:46. 
     
     
         7 . The nucleic acid construct according to  claim 1 , wherein the DNA promoter sequence is a tissue-specific promoter. 
     
     
         8 . The nucleic acid construct according to  claim 1 , wherein the DNA promoter sequence is an organ-specific promoter. 
     
     
         9 . An expression vector comprising the nucleic acid construct according to  claim 1 . 
     
     
         10 . A host cell transformed with the nucleic acid construct according to  claim 1 . 
     
     
         11 . The host cell according to  claim 10 , wherein the host cell is a bacterial cell or a plant cell. 
     
     
         12 . The host cell according to  claim 11 , wherein the host cell is a plant cell. 
     
     
         13 . A plant transformed with the nucleic acid construct according to  claim 1 . 
     
     
         14 . The plant according to  claim 13 , wherein the plant is selected from the group consisting of nice, corn, soybean, canola, potato, wheat, mung bean, alfalfa, barley, rye, cotton, sunflower, peanut, sweet potato, bean, pea, chicory, lettuce, endive, cabbage, brussel sprout, beet, parsnip, turnip, cauliflower, broccoli, radish, spinach, onion, garlic, eggplant, pepper, celery, carrot, squash, pumpkin, zucchini, cucumber, apple, pear, melon, citrus, strawberry, grape, raspberry, pineapple, tobacco, tomato, sorghum, sugarcane, banana,  Arabidopsis thaliana, Saintpaulia,  petunia, pelargonium, poinsettia, chrysanthemum, carnation, crocus, marigold, daffodil, pine,  Medicago truncatula, Sandersonia aurantiaca,  and zinnia. 
     
     
         15 . A component part of the plant according to  claim 13 . 
     
     
         16 . A fruit of the plant according to  claim 13 . 
     
     
         17 . A plant seed produced from the plant according to  claim 13 . 
     
     
         18 . A plant seed transformed with the nucleic acid construct according to  claim 1 . 
     
     
         19 . A method of expressing a nucleic acid molecule modulated by nitrogen in a plant, said method comprising:
 providing a transgenic plant or plant seed transformed with a nucleic acid construct according to  claim 1  and growing the transgenic plant or a plant grown from the transgenic plant seed under conditions effective to express the nucleic acid molecule in said transgenic plant or said plant grown from the transgenic plant seed.   
     
     
         20 . The method according to  claim 19 , wherein said growing is effective in reducing nitrogen uptake of said transgenic plant or said plant grown from the transgenic plant seed. 
     
     
         21 . The method according to  claim 19 , wherein said growing is effective in increasing nitrogen uptake of said transgenic plant or said plant grown from the transgenic plant seed. 
     
     
         22 . The method according to  claim 19 , wherein said growing is effective in increasing efficiency of nitrogen utilization of said transgenic plant or said plant grown from the transgenic plant seed. 
     
     
         23 . The method according to  claim 19 , wherein a transgenic plant is provided. 
     
     
         24 . The method according to  claim 19 , wherein a transgenic plant seed is provided. 
     
     
         25 . The method according to  claim 19 , wherein the plant is selected from the group consisting of rice, corn, soybean, canola, potato, wheat, mung bean, alfalfa, barley, rye, cotton, sunflower, peanut, sweet potato, bean, pea, chicory, lettuce, endive, cabbage, brussel sprout, beet, parsnip, turnip, cauliflower, broccoli, radish, spinach, onion, garlic, eggplant, pepper, celery, carrot, squash, pumpkin, zucchini, cucumber, apple, pear, melon, citrus, strawberry, grape, raspberry, pineapple, tobacco, tomato, sorghum, sugarcane, banana,  Arabidopsis thaliana, Saintpaulia,  petunia, pelargonium, poinsettia, chrysanthemum, carnation, crocus, marigold, daffodil, pine,  Medicago truncatula, Sandersonia aurantiaca,  and zinnia. 
     
     
         26 . The method according to  claim 19 , wherein said providing comprises transforming a non-transgenic plant or a non-transgenic plant seed with the nucleic acid construct to yield said transgenic plant or plant seed. 
     
     
         27 . The method according to  claim 26 , wherein said transforming comprises  Agrobacterium -mediated transformation, whisker method transformation, vacuum infiltration, biolistic transformation, electroporation, micro-injection, polyethylene-mediated transformation, or laser-beam transformation. 
     
     
         28 . An isolated DNA promoter from corn suitable for inducing nitrogen-regulated expression of a protein encoded by an isolated DNA molecule operably associated with the DNA promoter, wherein said DNA promoter comprises a nucleotide sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:6, SEQ ID NO:9, SEQ ID NO:12, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:21, SEQ ID NO:26, SEQ ID NO:29, SEQ ID NO:33, SEQ ID NO:38, SEQ ID NO:41, and SEQ ID NO:46. 
     
     
         29 . A nucleic acid construct comprising:
 an isolated nucleic acid molecule encoding a protein;   an isolated DNA promoter according to  claim 28 , wherein the DNA promoter is operably linked 5′ to the isolated nucleic acid molecule to induce transcription of the nucleic acid molecule; and   a 3′ terminator sequence operably linked to the isolated nucleic acid molecule.   
     
     
         30 . An expression vector comprising the nucleic acid construct according to  claim 29 . 
     
     
         31 . A host cell transformed with the nucleic acid construct according to  claim 29 . 
     
     
         32 . A plant transformed with the nucleic acid construct according to  claim 29 . 
     
     
         33 . A plant seed produced from the plant according to  claim 32 . 
     
     
         34 . A plant seed transformed with the nucleic acid construct according to  claim 29 . 
     
     
         35 . A method of directing nitrogen-regulated expression of an isolated nucleic acid in plants, said method comprising:
 transforming a plant cell with a nucleic acid construct according to  claim 29  and   regenerating a plant from the transformed plant cell, wherein expression of the nucleic acid molecule, under control of the DNA promoter, occurs in the plant and is upregulated by nitrogen.   
     
     
         36 . An isolated nucleic acid molecule that is modulated by nitrogen in corn, wherein said nucleic acid molecule comprises:
 a nucleotide sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:10, SEQ ID NO:13, SEQ ID NO:16, SEQ ID NO:19, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:27, SEQ ID NO:30, SEQ ID NO:34, SEQ ID NO:36, SEQ ID NO:39, SEQ ID NO:42, and SEQ ID NO:44; or   a coding portion of a nucleotide sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:10, SEQ ID NO:13, SEQ ID NO:16, SEQ ID NO:19, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:27, SEQ ID NO:30, SEQ ID NO:34, SEQ ID NO:36, SEQ ID NO:39, SEQ ID NO:42, and SEQ ID NO:44.   
     
     
         37 . An isolated protein or polypeptide, wherein said isolated protein or polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:5, SEQ ID NO:8, SEQ ID NO:11, SEQ ID NO:14, SEQ ID NO:17, SEQ ID NO:20, SEQ ID NO:23, SEQ ID NO:25, SEQ ID NO:28, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:35, SEQ ID NO:37, SEQ ID NO:40, SEQ ID NO:43, and SEQ ID NO:45.