Plants with Improved Nitrogen Utilization and Stress Tolerance

Abstract

The present invention relates to transgenic plants that have increased nitrogen use efficiency, stress tolerance, or both and that have been transformed using a novel vector construct including nucleic acid sequences that modulate nitrogen use in plants. In various embodiments, the vector construct comprises a nucleic acid sequence selected from the group consisting of SEQ ID NO: 2, 4, 7, 9, 11, 13, 15, 17, 22, 24, 26, 28, 30, 32, 34, 36, or 38. The invention also relates to isolated vectors for transforming plants and to antibodies used for detecting transformed plants. The invention also relates to methods of expressing in plants the nucleic acid molecules corresponding to the nucleic acid sequences that modulate nitrogen use in plants or are modulated by nitrogen conditions.

1 . A method for increasing nitrogen utilization efficiency in a plant comprising:
 (a) transforming a plant cell with an expression vector comprising an isolated nucleic acid molecule that comprises a nucleotide sequence that encodes an amino acid sequence that is at least 95% identical to the sequence set forth in SEQ ID NO: 5,   (b) expressing said nucleic acid molecule in said plant cell;   (c) generating from said plant cell a transformed plant in which said nucleotide sequence is overexpressed; and   (d) selecting from a plurality of said transformed plants a plant having increased nitrogen use efficiency as compared to a control plant grown under the same conditions.   
     
     
         2 . The method according to  claim 1 , said expression vector further comprising a 5′ DNA promoter sequence and a 3′ terminator sequence, wherein the nucleotide sequence, the DNA promoter sequence, and the terminator sequence are operatively coupled to permit transcription of the nucleotide sequence. 
     
     
         3 . The method according to  claim 2 , wherein the promoter sequence is selected from the group consisting of constitutive plant promoters and tissue specific promoters. 
     
     
         4 . A plant, comprising a plant produced by the method of  claim 1 . 
     
     
         5 . A plant according to  claim 4 , wherein the plant is selected from the group consisting of corn (maize); sorghum; wheat; sunflower; crucifers; peppers; potato; cotton; rice; soybean; sugarbeet; sugarcane; tobacco; barley; oilseed rape;  Brassica  sp.; alfalfa; rye; millet; safflower; peanuts; sweet potato; cassaya; coffee; coconut; pineapple; citrus trees; cocoa; tea; banana; avocado; fig; guava; mango; olive; papaya; cashew;  macadamia ; almond; oats; vegetables; grasses; azalea, hydrangea, hibiscus, roses, tulips, daffodils, petunias, carnation, poinsettia, and chrysanthemum; and conifers. 
     
     
         6 . Transgenic plant tissue of a plant of  claim 5 . 
     
     
         7 . Transgenic plant seed produced from a plant of  claim 5 . 
     
     
         8 . A host cell, comprising a host cell transformed with at least a first nucleotide sequence using the method of  claim 1 . 
     
     
         9 . The host cell according to  claim 8 , wherein the host cell is selected from the group consisting of bacterial cells and plant cells. 
     
     
         10 . A method of expressing a nucleic acid molecule modulated by nitrogen in a plant, said method comprising the steps of providing a transgenic plant or plant seed transformed with a vector 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. 
     
     
         11 . A method according to  claim 10 , wherein expression of the nucleic acid molecule is effective in increasing nitrogen uptake of said transgenic plant or said plant grown from the transgenic plant seed. 
     
     
         12 . A method according to  claim 10 , wherein expression of the nucleic acid molecule is effective in increasing efficiency of nitrogen utilization of said transgenic plant or said plant grown from the transgenic plant seed. 
     
     
         13 . A method according to  claim 10 , wherein the plant is selected from the group consisting of rice, corn, soybean, canola, wheat, alfalfa, barley, rye, cotton, sunflower, peanut, sweet potato, bean, pea, potato, oilseed rape, sorghum, forage grass, pasture grass, turf grass, sugarcane. 
     
     
         14 . A method of according to  claim 10 , wherein expression of the nucleic acid molecule is effective in improving the stress tolerance of said transgenic plant or said plant grown from the transgenic plant seed. 
     
     
         15 . A method according to  claim 10 , wherein expression of the nucleic acid molecule is effective in altering the morphology of said transgenic plant or said plant grown from the transgenic plant seed. 
     
     
         16 . A method for increasing nitrogen utilization efficiency in a plant comprising:
 (a) transforming a plant cell with an expression vector comprising an isolated nucleic acid molecule that comprises a nucleotide sequence that encodes an amino acid sequence set forth in SEQ ID NO: 5,   (b) expressing said nucleic acid molecule in said plant cell;   (c) generating from said plant cell a transformed plant in which said nucleotide sequence is overexpressed; and   (d) selecting from a plurality of said transformed plants a plant having increased nitrogen use efficiency as compared to a control plant grown under the same conditions.