Novel eto1 genes and use of same for reduced ethylene and improved stress tolerance in plants

Abstract

The invention provides isolated ethylene over-producer 1 (ETO1) nucleic acid molecules which are associated with ethylene production in plants and their encoded proteins. The present invention provides methods and compositions relating to altering ethylene production and abiotic stress response in plants. The invention further provides recombinant expression cassettes, host cells, transgenic plants, and antibody compositions.

1 . An isolated nucleic acid molecule comprising a member selected from the group consisting of:
 (a) a polynucleotide comprising a nucleotide sequence selected from the group consisting of the nucleotide sequences set forth in SEQ ID NO: 1, 3, 5, 7, and 9.   (b) a polynucleotide having at least 80% sequence identity, as determined by the BLAST 2.0 algorithm under default parameters, to a nucleotide sequence selected from the group consisting of the nucleotide sequences set forth in SEQ ID NO: 1, 3, 5, 7, and 9, wherein the polynucleotide encodes a polypeptide comprising ETO1 activity;   (c) a polynucleotide encoding a polypeptide comprising an amino acid selected from the selected from the group consisting the amino acid sequences set forth in SEQ ID NO: 2, 4, 6, 8, and 10;   (d) a polynucleotide amplified from a nucleic acid library using primers which selectively hybridize, under stringent hybridization conditions, to loci within a polynucleotide selected from the group consisting the nucleotide sequences set forth in SEQ ID NO: 1, 3, 5, 7, and 9, wherein the polynucleotide encodes a polypeptide comprising ETO1 activity;   (e) a polynucleotide which selectively hybridizes, under stringent hybridization conditions and a wash in 2× SSC at 50° C., to a polynucleotide selected from the group consisting of the nucleotide sequences set forth in SEQ ID NO: 1, 3, 5, 7, and 9, wherein the polynucleotide encodes a polypeptide comprising ETO1 activity;   (f) a polynucleotide comprising a nucleotide sequence encoding an amino acid sequence having at least 80% sequence identity to an amino acid selected from the selected from the group consisting the amino acid sequences set forth in SEQ ID NO: 2, 4, 6, 8, and 10, wherein the polynucleotide encodes a polypeptide comprising ETO1 activity.   (g) a polynucleotide which is the full-length complement of a polynucleotide of (a), (b), (c), (d), (e), or (f); and   (h) a polynucleotide comprising at least 25 contiguous nucleotides from a polynucleotide of (a), (b), (c), (d), (e) ,or (f), wherein the polynucleotide encodes a polypeptide comprising ETO1 activity.   
     
     
         2 . The nucleic acid molecule of  claim 1 , wherein the nucleic acid molecule comprises a nucleotide sequence selected from the group consisting of the nucleotide sequences set forth in SEQ ID NO: 1, 3, 5, 7, and 9. 
     
     
         3 . The nucleic acid molecule of  claim 1 , wherein the nucleic acid molecule encodes a polypeptide comprising an amino acid selected from the selected from the group consisting the amino acid sequences set forth in SEQ ID NO: 2, 4, 6, 8, and 10. 
     
     
         4 . A recombinant expression cassette comprising the nucleic acid molecule of  claim 1  operably linked, in sense or anti-sense orientation, to a promoter. 
     
     
         5 . The recombinant expression cassette of  claim 4 , wherein the promoter is capable of driving the expression for an operably linked polynucleotide in a plant cell. 
     
     
         6 . A host cell comprising the recombinant expression cassette of  claim 4 . 
     
     
         7 . A transgenic plant comprising the recombinant expression cassette of  claim 4 . 
     
     
         8 . The transgenic plant of  claim 7 , wherein the promoter is capable of driving the expression for an operably linked polynucleotide in a plant cell. 
     
     
         9 . The transgenic plant of  claim 7 , wherein said plant is a monocot or a dicot. 
     
     
         10 . The transgenic plant of  claim 7 , wherein said plant is selected from the group consisting of: maize, soybean, sunflower, sorghum, canola, wheat, alfalfa, cotton, rice, barley, and millet. 
     
     
         11 . A seed of the plant of  claim 7 , wherein the seed comprises the recombinant expression cassette. 
     
     
         12 . A method of modulating the ethylene production in a plant, comprising:
 (a) introducing into a plant cell a recombinant construct comprising a polynucleotide of  claim 1  operably linked to a promoter that is capable of driving the expression of the polynucleotide in a plant cell;   (b) culturing the plant cell under plant cell growing conditions and regenerating a plant therefrom; and   (c) inducing expression of said polynucleotide for a time sufficient to modulate the level of ethylene synthesis in said plant.   
     
     
         13 . The method of  claim 12 , wherein the plant is maize. 
     
     
         14 . A method of modulating the ethylene production in a plant, comprising:
 (a) introducing into a plant cell a nucleotide construct comprising a polynucleotide which encodes an ETO1 protein operably linked to a promoter;   (b) culturing the plant cell under plant cell growing conditions; and   (c) regenerating a plant from said plant cell; wherein the ethylene production in said plant is modulated.   
     
     
         15 . The method of  claim 14 , wherein said plant cell is from a monocot or a dicot. 
     
     
         16 . The method of  claim 14 , wherein the plant is selected from the group consisting of: maize, soybean, sorghum, canola, wheat, alfalfa, cotton, rice, barley, millet, peanut, and cocoa. 
     
     
         17 . The method of  claim 14 , wherein the ethylene production is increased or decreased. 
     
     
         18 . The method of  claim 14 , wherein the ethylene production is decreased and the construct is an over-expression construct. 
     
     
         19 . The method of  claim 18 , wherein the over-expression construct increases the presence of ETO1 proteins. 
     
     
         20 . The method of  claim 14 , wherein the polynucleotide comprises a nucleotide sequence selected from the group consisting of the nucleotide sequences set forth in SEQ ID NO: 1, 3, 5, 7 and 9. 
     
     
         21 . A transgenic plant produced by the method of  claim 14 . 
     
     
         22 . The transgenic plant of  claim 21 , wherein the plant has decreased ethylene production when compared to a control plant. 
     
     
         23 . An isolated protein comprising a member selected from the group consisting of:
 (a) a polypeptide comprising an amino acid sequence selected from the group consisting of the amino acid sequence set forth in SEQ ID NO: 2, 4, 6, 8 and 10;   (b) a polypeptide having at least 80% sequence identity to a polypeptide comprising an amino acid sequence selected from the group consisting of the amino acid sequences set forth in SEQ ID NO: 2, 4, 6, 8 and 10, wherein said sequence identity is determined using BLAST 2.0 under default parameters wherein the polypeptide comprises ETO1 activity; and   (c) a polypeptide having at least 80% sequence identity to, and having at least one linear epitope in common with, a polypeptide comprising an amino acid sequence selected from the group consisting of the amino acid sequences set forth in SEQ ID NO: 2, 4, 6, 8 and 10, wherein said sequence identity is determined using BLAST 2.0 under default parameters, wherein the polypeptide comprises ETO1 activity; and   (d) a polypeptide comprising at least 20 contiguous amino acids from a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 2, 4, 6, 8 and 10, wherein the polypeptide comprises ETO1 activity; and   (e) a polypeptide encoded by a nucleotide sequence selected from the group consisting of the nucleotide sequences set forth in SEQ ID NO: 1, 3, 5, 7, and 9.   
     
     
         24 . The protein of  claim 23 , wherein the protein comprises an amino acid sequence selected from the group consisting of the amino acid sequence set forth in SEQ ID NO: 2, 4, 6, 8 and 10.