Transgenic plants with enhanced agronomic traits

Abstract

This disclosure describes screening a population of transgenic plants derived from plant cells transformed with recombinant DNA for expression of proteins with homeobox domains to identify plant cells of specific transgenic events that are useful for imparting enhanced traits to transgenic crop plants. Traits include enhanced nitrogen use efficiency, increased yield, enhanced water use efficiency, enhanced tolerance to cold stress and/or improved seed compositions. Also disclosed are transgenic seeds for growing a transgenic plant having the recombinant DNA in its genome and exhibiting the screened enhance trait. Also disclosed are methods for generating seed and plants based on the transgenic events.

What is claimed is: 
     
         1 . A plant cell with stably integrated, recombinant DNA comprising a promoter that is functional in plant cells and that is operably linked to DNA that encodes a protein having domains of amino acids in a sequence that exceed the Pfam gathering cutoff for amino acid sequence alignment with a Pfam Homeobox protein domain family and a Pfam HALZ protein domain family; wherein the Pfam gathering cuttoff for the Homeobox protein domain family is −4 and the and the Pfam gathering cuttoff for the HALZ protein domain family is 17; wherein said plant cell is selected from a population of plant cells with said recombinant DNA by screening plants that are regenerated from plant cells in said population and that express said protein for an enhanced trait as compared to control plants that do not have said recombinant DNA; and wherein said enhanced trait is selected from group of enhanced traits consisting of enhanced water use efficiency, enhanced cold tolerance, increased yield, enhanced nitrogen use efficiency, enhanced seed protein and enhanced seed oil. 
     
     
         2 . A plant cell of  claim 1  wherein said protein has an amino acid sequence with at least 90% identity to a consensus amino acid sequence in the group of consensus amino acid sequences consisting of the consensus amino acid sequence constructed from an alignment of SEQ ID NO:5 through SEQ ID NO:8. 
     
     
         3 . A plant cell of  claim 1  wherein said protein is selected from the group of proteins having an amino acid sequence of SEQ ID NO:5 through SEQ ID NO:8. 
     
     
         4 . A plant cell of  claim 1  further comprising DNA expressing a protein that provides tolerance from exposure to an herbicide applied at levels that are lethal to a wild type of said plant cell. 
     
     
         5 . A plant cell of  claim 4  wherein the agent of said herbicide is a glyphosate, dicamba, or glufosinate compound. 
     
     
         6 . A transgenic plant comprising a plurality of the plant cell of  claim 1   
     
     
         7 . A transgenic plant of  claim 6  which is homozygous for said recombinant DNA. 
     
     
         8 . A transgenic seed comprising a plurality of the plant cell of  claim 1 . 
     
     
         9 . A transgenic seed of  claim 8  from a corn, soybean, cotton, canola, alfalfa, wheat or rice plant. 
     
     
         10 . Non-natural, transgenic corn seed of  claim 9  wherein said seed can produce corn plants that are resistant to disease from the Mal de Rio Cuarto virus or the  Puccina sorghi  fungus or both. 
     
     
         11 . A transgenic pollen grain comprising a haploid derivative of a plant cell of  claim 1 . 
     
     
         12 . A method for manufacturing non-natural, transgenic seed that can be used to produce a crop of transgenic plants with an enhanced trait resulting from expression of stably-integrated, recombinant DNA comprising a promoter that is (a) functional in plant cells and (b) is operably linked to operably linked to DNA that encodes a protein having domains of amino acids in a sequence that exceed the Pfam gathering cutoff for amino acid sequence alignment with a Pfam Homeobox protein domain family and a Pfam HALZ protein domain family; wherein the Pfam gathering cuttoff for the Homeobox protein domain family is −4 and the and the Pfam gathering cuttoff for the HALZ protein domain family is 17; and wherein said enhanced trait is selected from the group of enhanced traits consisting of enhanced water use efficiency, enhanced cold tolerance, increased yield, enhanced nitrogen use efficiency, enhanced seed protein and enhanced seed oil, said method for manufacturing said seed comprising:
 (a) screening a population of plants for said enhanced trait and said recombinant DNA, wherein individual plants in said population can exhibit said trait at a level less than, essentially the same as or greater than the level that said trait is exhibited in control plants which do not express the recombinant DNA, 
 (b) selecting from said population one or more plants that exhibit the trait at a level greater than the level that said trait is exhibited in control plants, 
 (c) verifying that said recombinant DNA is stably integrated in said selected plants, 
 (d) analyzing tissue of a selected plant to determine the production of a protein having the function of a protein encoded by nucleotides in a sequence of one of SEQ ID NO:1-4; and 
 (e) collecting seed from a selected plant. 
 
     
     
         13 . A method of  claim 12  wherein plants in said population further comprise DNA expressing a protein that provides tolerance to exposure to an herbicide applied at levels that are lethal to wild type plant cells, and wherein said selecting is effected by treating said population with said herbicide. 
     
     
         14 . A method of  claim 13  wherein said herbicide comprises a glyphosate, dicamba, or glufosinate compound. 
     
     
         15 . A method of  claim 12  wherein said selecting is effected by identifying plants with said enhanced trait. 
     
     
         16 . A method of  claim 12  wherein said seed is corn, soybean, cotton, alfalfa, wheat or rice seed. 
     
     
         17 . A method of producing hybrid corn seed comprising:
 (a) acquiring hybrid corn seed from a herbicide tolerant corn plant which also has stably-integrated, recombinant DNA comprising a promoter that is (a) functional in plant cells and (b) is operably linked to operably linked to DNA that encodes a protein having domains of amino acids in a sequence that exceed the Pfam gathering cutoff for amino acid sequence alignment with a Pfam Homeobox protein domain family and a Pfam HALZ protein domain family; wherein the Pfam gathering cuttoff for the Homeobox protein domain family is −4 and the and the Pfam gathering cuttoff for the HALZ protein domain family is 17;   (b) producing corn plants from said hybrid corn seed, wherein a fraction of the plants produced from said hybrid corn seed is homozygous for said recombinant DNA, a fraction of the plants produced from said hybrid corn seed is hemizygous for said recombinant DNA, and a fraction of the plants produced from said hybrid corn seed has none of said recombinant DNA;   (c) selecting corn plants which are homozygous and hemizygous for said recombinant DNA by treating with an herbicide;   (d) collecting seed from herbicide-treated-surviving corn plants and planting said seed to produce further progeny corn plants;   (e) repeating steps (c) and (d) at least once to produce an inbred corn line;   (f) crossing said inbred corn line with a second corn line to produce hybrid seed.   
     
     
         18 . The method of selecting a plant comprising cells of  claim 1  wherein an immunoreactive antibody is used to detect the presence of said protein in seed or plant tissue. 
     
     
         19 . Anti-counterfeit milled seed having, as an indication of origin, a plant cell of  claim 1 . 
     
     
         20 . A method of growing a corn, cotton or soybean crop without irrigation water comprising planting seed having plant cells of  claim 1  which are selected for enhanced water use efficiency. 
     
     
         21 . A method of  claim 20  comprising providing up to 300 millimeters of ground water during the production of said crop. 
     
     
         22 . A method of growing a corn, cotton or soybean crop without added nitrogen fertilizer comprising planting seed having plant cells of  claim 1  which are selected for enhanced nitrogen use efficiency.