Expression of sucrose phosphorylase in plants

Abstract

Introducing sucrose phosphorylase activity into plants by transformation with a gene for the enzyme increases the rate of sucrose hydrolysis, leading to increased starch, oil, and protein levels. The preferred gene is from Streptococcus mutans . Surprisingly, in potatoes transformed to express this gene in tubers, reduced bruise discoloration susceptibility and increased uniformity of starch deposition throughout the tuber are achieved.

1 . A method of producing a transgenic plant, comprising the steps of: 
 (a) inserting into the genome of a plant cell a recombinant, double-stranded DNA molecule comprising: 
 (i) a promoter which functions in cells of target plant tissue,  
 (ii) a structural DNA sequence that causes the production of an RNA sequence which encodes a sucrose phosphorylase enzyme,  
 (iii) a 3′ non-translated DNA sequence which functions in plant cells to cause transcriptional termination and the addition of polyadenylated nucleotides to the 3′ end of the RNA sequence;  
   (b) obtaining transformed plant cells; and    (c) regenerating from said transformed plant cells a genetically transformed plant, the genome of which contains said recombinant, double-stranded DNA molecule of step (a).    
     
     
         2 . The method of  claim 1 , wherein said DNA sequence encoding sucrose phosphorylase enzyme is obtained from  Streptococcus mutans.    
     
     
         3 . The method of  claim 2 , wherein said DNA sequence encoding sucrose phosphorylase has the sequence shown in SEQ ID NO: 5.  
     
     
         4 . The method of  claim 1 , wherein said genetically transformed plant exhibits a property selected from the group consisting of containing a modified carbohydrate content; increased polysaccharide (e.g. starch) content; enhanced yield; improved uniformity of the distribution of solids; and reduced susceptibility to bruising discoloration.  
     
     
         5 . The method of  claim 4 , wherein said property is containing a modified carbohyhdrate content.  
     
     
         6 . The method of  claim 5 , wherein said modified carbohydrate content is an increase in solids content.  
     
     
         7 . The method of  claim 6 , wherein said genetically transformed plant is selected from the group consisting of potato and tomato.  
     
     
         8 . The method of  claim 4 , wherein said property is improved uniformity of the distribution of solids.  
     
     
         9 . The method of  claim 8 , wherein said genetically transformed plant is selected from the group consisting of potato and sweet potato.  
     
     
         10 . The method of  claim 4 , wherein said property is reduced susceptibility to bruising discoloration.  
     
     
         11 . The method of  claim 10 , wherein said genetically transformed plant is selected from the group consisting of potato, banana, apple, wheat, grape, and peach.  
     
     
         12 . The method of  claim 4 , wherein said property is increased polysaccharide (e.g. starch) content.  
     
     
         13 . The method of  claim 12 , wherein said genetically transformed plant is selected from the group consisting of maize, wheat, rice, tomato, potato, sweet potato, peanut, barley, cotton, strawberry, raspberry, and cassava.  
     
     
         14 . The method of  claim 4 , wherein said property is enhanced yield.  
     
     
         15 . The method of  claim 14 , wherein said genetically transformed plant is selected from the group consisting of maize, wheat, rice, tomato, potato, sweet potato, peanut, barley, sugarbeet, sugarcane, apple, pear, orange, peach, grape, cotton, strawberry, raspberry, and cassava.  
     
     
         16 . The method of  claim 1 , wherein said plant cell is selected from the group consisting of a potato plant cell, a maize plant cell, a rice plant cell, a wheat plant cell, a tomato plant cell, a barley plant cell, a sugarbeet plant cell, a sweet potato plant cell, a peanut plant cell, a sugarcane plant cell, a grape plant cell, a pear plant cell, an apple plant cell, an orange plant cell, a cassava plant cell, a banana plant cell, a plantain plant cell, a cotton plant cell, a strawberry plant cell, a raspberry plant cell, and a peach plant cell.  
     
     
         17 . The method of  claim 16 , wherein said plant cell is a potato plant cell.  
     
     
         18 . The method of  claim 16 , wherein said plant cell is a maize plant cell.  
     
     
         19 . The method of  claim 16 , wherein said plant cell is selected from the group consisting of a wheat plant cell, a barley plant cell, a rice plant cell, and a tomato plant cell.  
     
     
         20 . A recombinant, double-stranded DNA molecule comprising in sequence: 
 (a) a promoter which functions in cells of target plant tissue;    (b) a structural DNA sequence that causes the production of an RNA sequence which encodes a sucrose phosphorylase enzyme; and    (c) a 3′ non-translated region which functions in plant cells to cause transcriptional termination and the addition of polyadenylated nucleotides to the 3′ end of the RNA sequence.    
     
     
         21 . The DNA molecule of  claim 20 , wherein said DNA sequence encoding sucrose phosphorylase enzyme is obtained from  Streptococcus mutans.    
     
     
         22 . The DNA molecule of  claim 21 , wherein said DNA sequence encoding sucrose phosphorylase has the sequence shown in SEQ ID NO: 5.  
     
     
         23 . The DNA molecule of  claim 20 , wherein said promoter is selected from the group consisting of a zein promoter, a patatin promoter, a rice glutelin promoter, the soybean 7s promoter, a promoter of a subunit of ADPglucose pyrophosphorylase, the TFM7 promoter, and the TFM9 promoter.  
     
     
         24 . A transformed plant cell comprising a recombinant, double-stranded DNA molecule comprising in sequence: 
 (a) a promoter which functions in said plant cell;    (b) a structural DNA sequence that causes the production of an RNA sequence which encodes a sucrose phosphorylase enzyme; and    (c) a 3′ non-translated region which functions in plant cells to cause transcriptional termination and the addition of polyadenylated nucleotides to the 3′ end of the RNA sequence.    
     
     
         25 . The plant cell of  claim 24 , wherein said DNA sequence encoding sucrose phosphorylase enzyme is from  Streptococcus mutans.    
     
     
         26 . The plant cell of  claim 25 , wherein said DNA sequence encoding sucrose phosphorylase has the sequence shown in SEQ ID NO: 5.  
     
     
         27 . The plant cell of  claim 24 , wherein said promoter is selected from the group consisting of a zein promoter, a patatin promoter, a rice glutelin promoter, the soybean 7s promoter, a promoter of a subunit of ADPglucose pyrophosphorylase, the TFM7 promoter, and the TFM9 promoter.  
     
     
         28 . The plant cell of  claim 24 , wherein said plant cell is selected from the group consisting of a potato plant cell, a maize plant cell, a rice plant cell, a wheat plant cell, a tomato plant cell , a barley plant cell, a sugarbeet plant cell, a sweet potato plant cell, a peanut plant cell, a sugarcane plant cell, a grape plant cell, a pear plant cell, an apple plant cell, an orange plant cell, a cassava plant cell, a banana plant cell, a plantain plant cell, and a peach plant cell.