Method for speeding up plant growth and improving yield by introducing phosphatases in transgenic plant

Abstract

Transgenic plants having increased growth rate, increased sugar content, and increase yield are disclosed, and methods for making the same. The transgenic plants have a gene coding for a phosphatase having a C-terminal motif under control of a heterologous promoter incorporated into the genomic DNA of the plant.

1 . A method to make a transgenic plant having increased rate of plant growth and elevate plant yields comprising:
 introducing a gene coding for a phosphatase into a plant, the phosphatase gene encodes for a polypeptide comprising a C-terminal motif having the sequence of SEQ ID NO: 66 or a homologue thereof, and one or more sequences selected from the group consisting of SEQ ID NOS: 48-53.   
     
     
         2 . The method of  claim 1 , wherein the phosphatase gene is a purple acid phosphatase gene comprising a nucleotide sequence encoding a polypeptide comprising an amino acid sequence of one or more selected from SEQ ID NOS: 2, 6, 8, 19, 21, 23, 25, 27, 29, 31, 33, 35, and 47, and homologues thereof. 
     
     
         3 . The method of  claim 1 , wherein the phosphatase gene is a purple acid phosphatase gene comprising a nucleotide sequence selected from SEQ ID NOS: 1, 5, 7, 18, 20, 22, 24, 26, 28, 30, 32, 34, and 46 and homologues thereof. 
     
     
         4 . The method of  claim 2 , wherein said nucleotide sequence comprises one or more selected from the group consisting of SEQ ID NOs 1, 5, 7, 18, 20, 22, 24, 26, 28, 30, 32, 34, and 46 and homologues thereof. 
     
     
         5 . The method of  claim 2 , wherein said nucleotide sequence comprises a nucleotide sequence encoding a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 2 and conservatively substituted variants thereof. 
     
     
         6 . The method of  claim 2 , wherein said nucleotide sequence comprises a nucleotide sequence encoding a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOS: 54 and 55 and conservatively substituted variants thereof. 
     
     
         7 . The method of  claim 6 , wherein said nucleotide sequence comprises a nucleotide sequence encoding a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOS: 50 and 51. 
     
     
         8 . The method of  claim 2 , wherein said nucleotide sequence comprises a nucleotide sequence encoding a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOS: 57-60 and conservatively substituted variants thereof. 
     
     
         9 . The method of  claim 8 , wherein said nucleotide sequence comprises a nucleotide sequence encoding a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOS: 50 and 51. 
     
     
         10 . The method of  claim 2 , wherein said nucleotide sequence comprises a nucleotide sequence encoding a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOS: 61-64. 
     
     
         11 . The method of  claim 1 , wherein introducing the phosphatase gene up-regulates the enzymatic activity of sucrose phosphate synthase in the transgenic plant relative to the wild-type plant. 
     
     
         12 . The method of  claim 1 , wherein introducing the phosphatase gene up-regulates the sucrose and/or glucose level in the transgenic plant relative to the wild-type plant. 
     
     
         13 . The method of  claim 1 , wherein introducing the phosphatase gene increases the growth rate of the transgenic plant relative to the wild-type plants. 
     
     
         14 . The method of  claim 1 , wherein introducing the phosphatase gene results in a higher crop yield of the transgenic plant relative to the wild-type plants. 
     
     
         15 . The method of  claim 1 , wherein the plant is a species selected from one of the group consisting of the following genera:  Asparagus, Bromus, Hemerocalli, Hordeum, Loliu, Panicum, Pennisetum, Saccharum, Sorghum, Trigonell, Triticum, Zea, Antirrhinum, Arabidopsis, Arachis, Atropa, Brassica, Browallia, Capsicum, Carthamus, Cichorium, Citrus, Chrysanthemum, Cucumis, Datura, Daucus, Digitalis, Fragaria, Geranium, Glycine, Helianthus, Hyscyamus, Ipomoea, Latuca, Linum, Lotus, Solanum lycopersicon, Majorana, Malva, Gossypium, Manihot, Medicago, Nemesia, Nicotiana, Onobrychis, Pelargonium, Petunia, Ranunculus, Raphanus, Salpiglossis, Senecio, Sinapis, Solanum, Trifolium, Vigna , and  Vitis.    
     
     
         16 . The method of  claim 1 , wherein the plant is a species selected from the family  Brassica.    
     
     
         17 . A transformed plant, comprising:
 a plant comprising at least one additional gene coding for a phosphatase relative to a corresponding wild-type plant, the phosphatase gene encodes for a polypeptide comprising a C-terminal motif having the sequence of SEQ ID NO: 66 or a homologue thereof, and one or more sequences selected from the group consisting of SEQ ID NOS: 48-53.   
     
     
         18 . The transformed plant of  claim 17 , wherein the phosphatase gene is a purple acid phosphatase gene comprising a nucleotide sequence encoding a polypeptide comprising an amino acid sequence of one or more selected from SEQ ID NOS: 2, 6, 8, 19, 21, 23, 25, 27, 29, 31, 33, 35, and 47, and homologues thereof. 
     
     
         19 . The transgenic plant of  claim 17 , wherein the phosphatase gene is a purple acid phosphatase gene comprising a nucleotide sequence selected from SEQ ID NOS: 1, 5, 7, 18, 20, 22, 24, 26, 28, 30, 32, 34, and 46 and homologues thereof. 
     
     
         20 . The transgenic plant of  claim 17 , wherein the phosphatase gene is a purple acid phosphatase gene comprising a nucleotide sequence selected from SEQ ID NO: 1 or a homologue thereof. 
     
     
         21 . The transformed plant of  claim 17 , wherein the phosphatase gene is a purple acid phosphatase gene comprising a nucleotide sequence encoding a polypeptide comprising an amino acid sequence of one or more selected from SEQ ID NOS: 2 or a homologue thereof. 
     
     
         22 . The transformed plant of  claim 17 , wherein said plant is of a monocotyledonous species. 
     
     
         23 . The transformed plant of  claim 17 , wherein said plant is of a dicotyledonous species. 
     
     
         24 . The transformed plant of  claim 17 , wherein the plant is a species selected from one of the group consisting of the following genera:  Asparagus, Bromus, Hemerocalli, Hordeum, Loliu, Panicum, Pennisetum, Saccharum, Sorghum, Trigonell, Triticum, Zea, Antirrhinum, Arabidopsis, Arachis, Atropa, Brassica, Browallia, Capsicum, Carthamus, Cichorium, Citrus, Chrysanthemum, Cucumis, Datura, Daucus, Digitalis, Fragaria, Geranium, Glycine, Helianthus, Hyscyamus, Ipomoea, Latuca, Linum, Lotus, Solanum lycopersicon, Majorana, Malva, Gossypium, Manihot, Medicago, Nemesia, Nicotiana, Onobrychis, Pelargonium, Petunia, Ranunculus, Raphanus, Salpiglossis, Senecio, Sinapis, Solanum, Trifolium, Vigna , and  Vitis.    
     
     
         25 . The transformed plant of  claim 17 , wherein the plant is a species selected from the family  Brassica.    
     
     
         26 . A vector comprising a plasmid comprising a phosphatase gene comprising a nucleic acid sequence selected from the group consisting of SEQ ID NOS: 1, 5, 7, 18, 20, 22, 24, 26, 28, 30, 32, 34, and 46 and homologues thereof, wherein the plasmid is capable of transforming a bacterial cell. 
     
     
         27 . The vector of  claim 26 , wherein the bacterial cell is  Agrobacterium tumefaciens.    
     
     
         28 . The vector of  claim 26 , wherein said phosphatase gene encodes an amino acid sequence comprising one or more selected from the group consisting of SEQ ID NOS: 2, 6, 8, 19, 21, 23, 25, 27, 29, 31, 33, 35, and 47 and homologues thereof, except that all or an N-terminal portion of amino acid residues 1 to 50 of these amino acid sequences are replaced by a plant signal peptide such that said polypeptide is sorted to various organelles or compartments of plant cells upon expression of the phosphatase gene in a transformed host plant cell. 
     
     
         29 . The vector of  claim 26 , wherein the phosphatase gene comprises a nucleic acid sequence selected from SEQ ID NO: 1 or a homologue thereof. 
     
     
         30 . A host cell comprising the vector of  claim 26 . 
     
     
         31 . The host cell of  claim 30 , wherein the phosphatase gene comprising the vector is operably linked to a heterologous promoter. 
     
     
         32 . The host cell of  claim 31 , wherein the heterologous promoter is a plant promoter. 
     
     
         33 . The host cell of  claim 31 , wherein the heterologous promoter is a promoter derived from cauliflower mosaic virus. 
     
     
         34 . The host cell of  claim 30 , wherein the host cell is a plant species. 
     
     
         35 . A method for preparing a cell or progeny thereof capable of expressing a purple acid phosphatase in a host cell, comprising:
 transforming a bacterial cell with the vector of  claim 26  and   transforming the host cell by transfer of DNA from the bacterial cell to the host cell.   
     
     
         36 . The method of  claim 35 , wherein the bacterial cell is  Agrobacterium tumefaciens  and the host cell is a plant cell. 
     
     
         37 . The method of  claim 36 , wherein the host cell is from the family  Brassica.    
     
     
         38 . An expression cassette comprising a phosphatase gene comprising a nucleotide sequence encoding an enzyme having phosphatase activity, wherein said nucleotide sequence hybridizes to a nucleotide sequences selected from the group consisting of SEQ ID NOS: 1, 5, 7, 18, 20, 22, 24, 26, 28, 30, 32, 34, and 46, under stringent condition and is operably linked to regulatory nucleotide sequences such that said regulatory nucleotide sequences cause expression of the nucleotide sequence in plant cells. 
     
     
         39 . The expression cassette of  claim 38 , wherein said nucleotide sequence hybridizes to the nucleotide sequence of SEQ ID NO: 1. 
     
     
         40 . The Expression cassette of  claim 38 , wherein said regulator nucleotide sequence is a cauliflower mosaic virus promoter. 
     
     
         41 . A method of feeding animals, comprising:
 providing a feed comprising matter derived from the transformed plant of  claim 17  to an animal for consumption by the animal.