Potatoes with reduced cold-induced sweetening

Abstract

Materials and methods are provided for making plants (e.g., Solanum varieties) with decreased accumulation of reducing sugars and acrylamide in cold-stored potatoes, specifically, by making TALE-nuclease-induced mutations in genes encoding vacuolar invertase.

1 . A  Solanum  plant, plant part, or plant cell comprising a mutation in at least two VInv alleles endogenous to said plant, plant part, or plant cell, such that said plant, plant part, or plant cell has reduced expression of vacuolar invertase as compared to a control  Solanum  plant, plant part, or plant cell that lacks said mutation. 
     
     
         2 . The plant, plant part, or plant cell of  claim 1 , wherein each said mutation is a deletion of more than one nucleotide base pair. 
     
     
         3 . The plant, plant part, or plant cell of  claim 1 , wherein each said mutation is at a target sequence as set forth in SEQ ID NO:27, or at a target sequence having at least 95 percent identity to SEQ ID NO:27. 
     
     
         4 . The plant, plant part, or plant cell of  claim 1 , wherein each said mutation is at a target sequence as set forth in SEQ ID NO:1, or at a target sequence having at least 95 percent identity to SEQ ID NO:1. 
     
     
         5 . The plant, plant part, or plant cell of  claim 1 , wherein said plant, plant part, or plant cell was made using a rare-cutting endonuclease. 
     
     
         6 . The plant, plant part, or plant cell of  claim 5 , wherein said rare-cutting endonuclease is a transcription activator-like effector endonuclease (TALE-nuclease). 
     
     
         7 . The plant, plant part, or plant cell of  claim 6 , wherein said TALE-nuclease binds to a sequence as set forth in any of SEQ ID NOS:18-23. 
     
     
         8 . The plant, plant part, or plant cell of  claim 1 , wherein each of said at least two VInv alleles exhibits removal of an endogenous nucleic acid and does not include any exogenous nucleic acid. 
     
     
         9 . The plant, plant part, or plant cell of  claim 1 , wherein every endogenous VInv allele is mutated. 
     
     
         10 . The plant, plant part, or plant cell of  claim 9 , wherein each said VInv allele exhibits removal of an endogenous nucleic acid and does not include any exogenous nucleic acid. 
     
     
         11 . The plant, plant part, or plant cell of  claim 9 , wherein said plant, plant part, or plant cell has no detectable expression of vacuolar invertase. 
     
     
         12 . The plant, plant part, or plant cell of  claim 1 , wherein said  Solanum  plant, plant part, or plant cell is a  S. tuberosum  plant, plant part, or plant cell. 
     
     
         13 . The plant, plant part, or plant cell of  claim 1 , wherein said plant, plant part, or plant cell is subjected to cold storage conditions. 
     
     
         14 . The plant, plant part, or plant cell of  claim 13 , wherein said plant, plant part, or plant cell has decreased levels of acrylamide as compared to a control plant, plant part, or plant cell that lacks said mutation. 
     
     
         15 . A method for making a  Solanum  plant that has reduced cold-induced sweetening, wherein said method comprises:
 (a) contacting a population of  Solanum  plant cells comprising a functional VInv allele with a rare-cutting endonuclease targeted to an endogenous VInv sequence,   (b) selecting, from said population, a cell in which at least two VInv alleles have been inactivated, and   (c) growing said selected plant cell into a  Solanum  plant, wherein said  Solanum  plant has reduced cold-induced sweetening as compared to a control  Solanum  plant in which said VInv alleles have not been inactivated.   
     
     
         16 . The method of  claim 15 , wherein said  Solanum  plant cells are protoplasts. 
     
     
         17 . The method of  claim 16 , comprising transforming said protoplasts with a nucleic acid encoding said rare-cutting endonuclease. 
     
     
         18 . The method of  claim 17 , wherein said nucleic acid is an mRNA. 
     
     
         19 . The method of  claim 17 , wherein said nucleic acid is contained within a vector. 
     
     
         20 . The method of  claim 15 , comprising introducing into said protoplasts a rare-cutting endonuclease protein. 
     
     
         21 - 48 . (canceled)