Transformed plant and method for producing exudate containing sugar using transformed plant

Abstract

The production of exudate containing sugar from a plant at a high concentration is provided. A nucleic acid encoding a transporter involved in sugar transportation having a certain consensus sequence derived from the amino acid sequences of the SWEET proteins classified in the clade III is introduced and/or expression of the protein is enhanced.

1 . A method for producing an exudate, comprising the steps of cultivating a transformed plant in which a nucleic acid encoding a transporter protein having a consensus sequence comprising the following amino acid sequence:
 (L/I/V/M/F)x(G/A)xx(I/L/V/M/F)xxxx(L/I/V/F)(A/S)(P/S) [SEQ ID NO: 1] (1-3aa) (P/S/T/A)T (F/L)xx(I/V)xxxKxxxxxxxxPYxxx(L/I)xxxx(L/I)x(I/L/M/V/F)xY(A/S/G) [SEQ ID NO: 2] (7-13aa)(I/L/V/M)(1-2aa)(I/V)Nxxxxxx(E/Q)xxYxxx(Y/F)xx(Y/F)(A/G/S) [SEQ ID NO: 3] (35-36aa)(R/Q/H)xxxxGx(V/I/L)xxxxx(V/M/L/I/F)xxxx(A/S/T)P(L/M)x(I/V)(I/M/V/L) [SEQ ID NO: 4] (2-7aa) (V/I) (V/I/M)x(T/S)x(S/N)xx(F/Y) (M/L) (P/S) (F/I/V/L)xLSxx(L/I) (T/V)xx(A/G)xx W(F/L)xYGxxxxDxx(V/I)xxPNxxGxx(F/L) (G/S)xxQ(M/I)x(L/M/I/V/F) (Y/H/F) [SEQ ID NO: 5] and being involved in sugar transportation is introduced and/or expression of the protein is enhanced; and collecting an exudate from the transformed plant.   
     
     
         2 . The method for producing an exudate according to  claim 1 , wherein the transformed plant is cultivated under conditions at a relative humidity of 80% RH or more. 
     
     
         3 . The method for producing an exudate according to  claim 1 , wherein the exudate is guttation. 
     
     
         4 . The method for producing an exudate according to  claim 1 , wherein the transporter protein is a protein in the clade III among the clades I to V of taxonomic groups based on the amino acid sequences of the SWEET proteins. 
     
     
         5 . The method for producing an exudate according to  claim 1 , wherein the transporter protein is a protein of any of the following (a) and (b):
 (a) a protein comprising any of the amino acid sequences set forth in SEQ ID NOs: 15 to 137;   (b) a protein having an amino acid sequence having an identity of 90% or more to an amino acid sequence set forth in any one of SEQ ID NOs: 15 to 137 and having transporter activity involved in sugar transportation.   
     
     
         6 . The method for producing an exudate according to  claim 1  wherein the consensus sequence comprises the following amino acid sequence: 
       
         
           
                 
               
                   [SEQ ID NO: 6] 
                 
                   G(L/I/V/F/M)xGx(I/V/L)(I/V/L)(S/T)xxxxL(A/S)P(L/ 
                 
                     
                 
                   V/I/M)(P/S/T/A)TFxx(I/V)x(K/R)xK(S/T)xxx(F/Y)x(S/ 
                 
                     
                 
                   A)xPYxx(A/S/T)LxSxxLx(L/I/M/V)(Y/F)Y(A/G) 
                 
                     
                 
                   [SEQ ID NO: 7] 
                 
                   (7-9 aa)(L/I)(I/V/L)(T/S)INxx(G/A)xx(I/V/M)(E/Q) 
                 
                     
                 
                   xxYxxx(F/Y)(L/I/V/F)x(Y/F)Ax(K/R/N)xxxxx(T/A) 
                 
                     
                 
                   [SEQ ID NO: 8] 
                 
                   (7-8 aa)(V/F/L/I/M)(18-19 aa)(R/Q/H)xxxxGx(I/V) 
                 
                     
                 
                   xxxxx(V/I/L/M)x(V/M)F(A/V)(A/S/T)PLx(I/V)(I/M/V/L) 
                 
                     
                 
                   xxV(I/V)(K/R/Q)(T/S)(K/R)S(V/A)x(F/Y)MP(F/I/L)xLS 
                 
                     
                 
                   (L/F/V)xL(T/V)(L/I)xAxxW(F/L)xYG(L/F)xxxDxx(V/I) 
                 
                     
                 
                   xxPNxxGxx(L/F)(G/S)xxQMx(L/V/I)(Y/F)xx(Y/F). 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         7 . The method for producing an exudate according to  claim 6 , wherein the transporter protein is a protein of any of the following (a) and (b):
 (a) a protein comprising any of the amino acid sequences set forth in SEQ ID NOs: 15 to 35;   (b) a protein having an amino acid sequence having an identity of 90% or more to an amino acid sequence set forth in any one of SEQ ID NOs: 15 to 35 and having transporter activity involved in sugar transportation.   
     
     
         8 . The method for producing an exudate according to  claim 1 , wherein the consensus sequence comprises the following amino acid sequence: 
       
         
           
                 
               
                   [SEQ ID NO: 9] 
                 
                   (A/V)xxxG(I/L/V)xGN(I/L/V)(I/L/V)S(F/L)x(V/T)xL 
                 
                     
                 
                   (A/S)P(V/L/I)(P/A)TFxx(I/V)x(K/R)xK(S/T)xx(G/S) 
                 
                     
                 
                   (F/Y)(Q/S/E)SxPYxx(A/S/T)LxS(A/C/S)xLx(L/I/M)(Y/F) 
                 
                     
                 
                   Y(A/G)xx(K/T) 
                 
                     
                 
                   [SEQ ID NO: 10] 
                 
                   (3-5 aa)(L/M/P)(L/I)(I/L/V)(T/S)INxx(G/A)xx(I/V) 
                 
                     
                 
                   (E/Q)xxY(I/L)x(L/M/V/I)(F/Y)(L/I/V/F)x(Y/F)Ax(K/R) 
                 
                     
                 
                   xxxxx(T/A)xx(L/M/F/V/I)(L/F/V/I)xxx(N/D)(F/V/I/L) 
                 
                     
                 
                   xx(F/L)xx(I/L/V)xxxxxx(L/I/V) 
                 
                     
                 
                   [SEQ ID NO: 11] 
                 
                   (5-6 aa)(R/Q)xxxxGx(I/V)xxxx(S/A)(V/L/M)(C/S/A)VF 
                 
                     
                 
                   (A/V)(A/S)P Lx(I/V)(I/M/V)xxV(I/V)(K/R/Q)(T/S)(K/ 
                 
                     
                 
                   R)S(V/A)E(F/Y)MP(F/I)xLS(L/F/V)xL(T/V)(L/I)(S/N)A 
                 
                     
                 
                   (V/I)xW(F/L)xYGLxx(K/N)Dxx(V/I)xxPN(V/I)xGxx(F/L) 
                 
                     
                 
                   (G/S)xxQMxL(Y/F)xx(Y/F). 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         9 . The method for producing an exudate according to  claim 8 , wherein the transporter protein is a protein of any of the following (a) and (b):
 (a) a protein comprising any of the amino acid sequences set forth in SEQ ID NOs: 15 to 26;   (b) a protein having an amino acid sequence having an identity of 90% or more to an amino acid sequence set forth in any one of SEQ ID NOs: 15 to 26 and having transporter activity involved in sugar transportation.   
     
     
         10 . The method for producing an exudate according to  claim 1 , wherein the consensus sequence comprises the following amino acid sequence: 
       
         
           
                 
               
                   [SEQ ID NO: 12] 
                 
                   (M/L/V)xx(T/K/N/S)xxxxAxxFG(L/I/V)LGN(I/L/V)(I/V) 
                 
                     
                 
                   SFxVxL(S/A)P(V/I)PTFxxIxK(K/R)K(S/T)x(E/K)(G/S)(F/ 
                 
                     
                 
                   Y)(Q/E)S(I/L)PYxx(A/S)LxS(A/C)xLx(L/I/M)YY(A/G)xxK 
                 
                     
                 
                   [SEQ ID NO: 13] 
                 
                   (4-5 aa)(L/M)(L/I)(I/V)(T/S)IN(A/S/T)(F/V)(G/A)x 
                 
                     
                 
                   (F/V)(I/V)(E/Q)xxY(I/L)x(L/M/I)(F/Y)(F/V/I/L)x(Y/ 
                 
                     
                 
                   F)Ax(K/R)xx(R/K)xx(T/A)(L/V/M)K(V/L/M/F)(L/I/V/F) 
                 
                     
                 
                   xxx(N/D)(F/V/I)xx(F/L)xx(I/L)(L/I/V/F)(L/M/V)(L/V) 
                 
                     
                 
                   xx(F/L)(L/I/V) 
                 
                     
                 
                   [SEQ ID NO: 14] 
                 
                   (5-6 aa)(R/Q)x(K/S/Q)x(L/I/V)Gx(I/V)Cxxx(S/A)(V/L) 
                 
                     
                 
                   (S/C/A)VF(A/V)(A/S)PLx(I/V)(M/I/V)xxV(I/V)(K/R)T 
                 
                     
                 
                   (K/R)S(V/A)E(Y/F)MPFxLS(L/F)xLT(I/L)(S/N)A(V/I)xW 
                 
                     
                 
                   (L/F)xYGLx(L/I)(K/N)Dxx(V/I)A(L/F/I/M)PN(V/I)(L/ 
                 
                     
                 
                   I/V)Gxx(L/F)GxxQM(I/V)L(Y/F)(V/L/I/M)(V/L/I/M)(Y/ 
                 
                     
                 
                   F)(K/R/Q). 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         11 . The method for producing an exudate according to  claim 10 , wherein the transporter protein is a protein of any of the following (a) and (b):
 (a) a protein comprising any of the amino acid sequences set forth in SEQ ID NOs: 15 to 21;   (b) a protein comprising an amino acid sequence having an identity of 90% or more to an amino acid sequence set forth in any one of SEQ ID NOs: 15 to 21.   
     
     
         12 . The method for producing an exudate according to  claim 1 , wherein the transformed plant is a phanerogam. 
     
     
         13 . The method for producing an exudate according to  claim 12 , wherein the phanerogam is an angiosperm. 
     
     
         14 . The method for producing an exudate according to  claim 13 , wherein the angiosperm is a monocot. 
     
     
         15 . The method for producing an exudate according to  claim 14 , wherein the monocot is a plant of the family Poaceae. 
     
     
         16 . The method for producing an exudate according to  claim 15 , wherein the plant of the family Poaceae is a plant of the genus  Oryza.    
     
     
         17 . The method for producing an exudate according to  claim 13 , wherein the angiosperm is a dicot. 
     
     
         18 . The method for producing an exudate according to  claim 17 , wherein the dicot is a plant of the family Brassicaceae. 
     
     
         19 . The method for producing an exudate according to  claim 18 , wherein the plant of the family Brassicaceae is a plant of the genus  Arabidopsis.