Method of constructing novel higher plant and method of promoting the growth of higher plant

Abstract

The present invention provides a method of producing a higher plant having cytochrome c 6 in the thylakoid space of chloroplast, which is characterized in that it comprises introducing a gene encoding a fused protein formed by adding a signal peptide consisting of 50 to 80 amino acid residues to a cytochrome c 6 protein into the genome of a higher plant.

1 . A method of producing a higher plant having cytochrome c 6  in the thylakoid space of chloroplast, comprising
 introducing a gene encoding a fused protein comprising a signal peptide consisting of 50 to 80 amino acid residues and a cytochrome c 6  protein into the genome of a higher plant,   wherein the cytochrome c 6  protein is from  Porphyra yezoensis.      
     
     
         2 . A method of promoting the growth of a higher plant, comprising
 introducing a gene encoding a fused protein comprising a signal peptide consisting of 50 to 80 amino acid residues and a cytochrome c 6  protein into the genome of a higher plant,   expressing the gene under suitable conditions,   wherein the cytochrome c 6  is located in the thylakoid space of chloroplast,   wherein the cytochrome c 6  protein is from  Porphyra yezoensis.      
     
     
         3 . A method of promoting the synthesis of at least one selected from the group consisting of the ATP, NADPH, starch, and protein of a higher plant, comprising
 introducing a gene encoding a fused protein comprising a signal peptide consisting of 50 to 80 amino acid residues and a cytochrome c 6  protein into the genome of a higher plant,   expressing the gene under suitable conditions,   wherein the cytochrome c 6  is located in the thylakoid space of chloroplast,   wherein the cytochrome c 6  protein is from  Porphyra yezoensis.      
     
     
         4 . A method of promoting carbon fixation by a higher plant, comprising
 introducing a gene encoding a fused protein formed by adding a signal peptide consisting of 50 to 80 amino acid residues and a cytochrome c 6  protein into the genome of a higher plant,   expressing the gene under suitable conditions,   wherein the cytochrome c 6  is located in the thylakoid space of chloroplast,   wherein the cytochrome c 6  protein is from  Porphyra yezoensis.      
     
     
         5 . The method according to any one of  claims 1  to  4 , wherein said fused protein is the protein described in the following (a), (b), (c), or (d):
 (a) a protein having the amino acid sequence as shown in SEQ ID NO: 6; 
 (b) a protein, which has an amino acid sequence comprising a deletion, substitution, or addition of one or several amino acids with respect to an amino acid sequence portion corresponding to said signal peptide in the amino acid sequence as shown in SEQ ID NO: 6, and which has ability to pass through the chloroplast envelope and thylakoid membrane of a higher plant; 
 (c) a protein, which has an amino acid sequence comprising a deletion, substitution, or addition of one or several amino acids with respect to an amino acid sequence portion corresponding to said cytochrome c 6  protein in the amino acid sequence as shown in SEQ ID NO: 6, and which has ability to transfer electrons; and 
 (d) a protein, which has an amino acid sequence comprising a deletion, substitution, or addition of one or several amino acids with respect to each of an amino acid sequence portion corresponding to said signal peptide and an amino acid sequence portion corresponding to said cytochrome c 6  protein in the amino acid sequence as shown in SEQ ID NO: 6, and which has ability to pass through the chloroplast envelope and thylakoid membrane of a higher plant and ability to transfer electrons. 
 
     
     
         6 . The method according to any one of  claims 1  to  4 , wherein said gene is a gene comprising the DNA described in the following (a) or (b):
 (a) DNA having the nucleotide sequence as shown in SEQ ID NO: 5; and 
 (b) DNA, which hybridizes with DNA having a nucleotide sequence complementary to the DNA having the nucleotide sequence as shown in SEQ ID NO: 5 under stringent conditions, and which encodes a protein having ability to pass through the chloroplast envelope and thylakoid membrane of a higher plant and ability to transfer electrons. 
 
     
     
         7 . A fused protein, comprising a signal peptide consisting of 50 to 80 amino acid residues and a cytochrome c 6  protein,
 wherein the cytochrome c 6  protein is from  Porphyra yezoensis.      
     
     
         8 . The fused protein according to  claim 7 , which is the protein described in the following (a), (b), (c), or (d):
 (a) a protein having the amino acid sequence as shown in SEQ ID NO: 6;   (b) a protein, which has an amino acid sequence comprising a deletion, substitution, or addition of one or several amino acids with respect to an amino acid sequence portion corresponding to said signal peptide in the amino acid sequence as shown in SEQ ID NO: 6, and which has ability to pass through the chloroplast envelope and thylakoid membrane of a higher plant;   (c) a protein, which has an amino acid sequence comprising a deletion, substitution, or addition of one or several amino acids with respect to an amino acid sequence portion corresponding to said cytochrome c 6  protein in the amino acid sequence as shown in SEQ ID NO: 6, and which has ability to transfer electrons; and   (d) a protein, which has an amino acid sequence comprising a deletion, substitution, or addition of one or several amino acids with respect to each of an amino acid sequence portion corresponding to said signal peptide and an amino acid sequence portion corresponding to said cytochrome c 6  protein in the amino acid sequence as shown in SEQ ID NO: 6, and which has ability to pass through the chloroplast envelope and thylakoid membrane of a higher plant and ability to transfer electrons.   
     
     
         9 . A gene, which encodes the fused protein according to  claim 7  or  8 . 
     
     
         10 . A gene, which comprises the DNA described in the following (a) or (b):
 (a) DNA having the nucleotide sequence as shown in SEQ ID NO: 5; and   (b) DNA, which hybridizes with DNA having a nucleotide sequence complementary to the DNA having the nucleotide sequence as shown in SEQ ID NO: 5 under stringent conditions, and which encodes a protein having ability to pass through the chloroplast envelope and thylakoid membrane of a higher plant and ability to transfer electrons.   
     
     
         11 . A recombinant vector comprising the gene according to  claim 9 . 
     
     
         12 . A transformant comprising the recombinant vector according to  claim 11 . 
     
     
         13 . The transformant according to  claim 12 , wherein the transformant is a microorganism belonging to genus  Agrobacterium.    
     
     
         14 . A transgenic higher plant, into the plant genome of which the gene according to  claim 9  has been introduced. 
     
     
         15 . The transgenic higher plant according to  claim 14 , which has cytochrome c 6  in the thylakoid space of chloroplast in a plant cell thereof. 
     
     
         16 . The method according to any one of  claims 1  to  4 , wherein the higher plant belongs to a plant family selected from the group consisting of solanaceae, gramineae, leguminosae, chenopodiaceae, rosaceae, asteraceae, liliaceae, caryophyllaceae, cucurbitaceae, convolvulaceae, amaranthaceae, bromeliaceae, cactaceae, aloeaceae and orchidaceae. 
     
     
         17 . The method according to any one of  claims 1  to  4 , wherein the higher plant is a C4 plant, a Crassulaean acid metabolism plant, or a C3-C4 intermediate plant. 
     
     
         18 . The method according to any one of  claims 1  to  4 , wherein the higher plant is a C3 plant selected from the group consisting of solanaceae, gramineae, leguminosae, chenopodiaceae, rosaceae, asteraceae, cucurbitaceae, convolvulaceae and orchidaceae. 
     
     
         19 . The transformant according to  claim 12 , wherein the transformant is selected from the group consisting of  Escherichia coli, Bacillus subtilis , a yeast cell and a mold cell.