Plastid transformation of solanaceous plants

Abstract

A method is provided for transforming solanaceous plants to express DNA sequences interest from the plant cell plastid. The improved method allows the transformation of solanaceous plant tissue which is not obtained from tobacco with DNA constructs. Such DNA constructs comprise, in the 5′ to 3′ direction of transcription, a promoter region functional in a plant plastid and a DNA sequence of interest. The method can be utilized in the transformation of solanaceous plants, such as potato and petunia. The invention further provides constructs and methods for the expression of green fluorescent protein from the plant cell plastid.

What is claimed is:  
     
         1 . In a method for transforming a plant cell plastid comprising the steps of introducing into cells of a plant a construct comprising a promoter functional in a plant cell plastid operably associated with a DNA sequence of interest and transforming said plant cell plastid with said construct, 
 the improvement comprising introducing said construct into a solanaceous plant cell which is other than a tobacco cell.    
     
     
         2 . The method according to  claim 1 , wherein said construct comprises regions of homology to the genome of said plastid whereby said construct is integrated into the plant cell plastid genome.  
     
     
         3 . The method according to  claim 2 , wherein said plastid homology regions are derived from a heterologous plastid genome.  
     
     
         4 . The method according to  claim 2 , wherein said plastid homology regions are derived from a homologous plastid genome.  
     
     
         5 . The method according to  claim 1  further comprising regenerating a mature plant containing transformed plastids from said plant cells.  
     
     
         6 . The method according to  claim 1  wherein said plant cell is a potato cell.  
     
     
         7 . The method according to  claim 6  wherein said plant cell comprises a leaf cell.  
     
     
         8 . The method according to  claim 7  comprising the step of wounding the surface of a potato leaf prior to introducing said construct in to said plant cell.  
     
     
         9 . The method according to  claim 1  wherein said plant cell is a petunia cell.  
     
     
         10 . The method according to  claim 9  wherein said plant cell comprises a leaf cell.  
     
     
         11 . The method according to  claim 1 , 
 wherein nuclei of said plant cells comprise a construct having the following as operably joined components in the 5′ to 3′ direction of transcription, a promoter functional in said nuclei, an encoding sequence for a plastid transit peptide, an encoding sequence for a viral single subunit RNA polymerase from a member of the T7 class of bacteriophage and a transcriptional termination region, and    wherein said promoter functional in said plant cell plastid is a promoter specific for said viral single subunit RNA polymerase.    
     
     
         12 . The method according to  claim 21 , wherein said promoter functional in said nuclei provides for tissue preferential or developmentally regulated expression.  
     
     
         13 . The method according to  claim 22  wherein said promoter functional in said nuclei is preferentially expressed in the tuber.  
     
     
         14 . The method according to  claim 23  wherein said promoter is selected from the group consisting of patatin, zein and plant starch synthase.  
     
     
         15 . A method for the expression of a green fluorescent protein marker in a plant cell plastid comprising transforming a plant cell plastid with a DNA construct with the following as operably linked components, in the 5′ to 3′ direction of transcription, (i) a promoter functional in a plant cell plastid, (ii) a DNA sequence encoding for a green fluorescent protein, and (iii) a transcriptional termination region functional in a plant cell plastid.  
     
     
         16 . The method according to  claim 1 , wherein said DNA sequence of interest is a DNA sequence which is capable of conferring tolerance in a plant cell to at least one herbicide compound when said DNA sequence is transcribed in plastids of said plant cell.  
     
     
         17 . The method according to  claim 1 , wherein said DNA sequence of interest encode genes providing for resistance to insects from  Bacillus thuringensis.    
     
     
         18 . The method according to  claim 1 , wherein said DNA sequence of interest encodes genes capable of directing the production of human biological proteins.  
     
     
         19 . A plant cell plastid obtained by the method of  claim 1 .  
     
     
         20 . A plant, plant seed, plant cell or progeny thereof containing a plant cell according to  claim 11 .  
     
     
         21 . A plant cell having plastids containing a heterologous construct, wherein said construct comprises, as operably linked components in the 5′ to 3′ direction of transcription, a promoter functional in a plant cell plastid, a DNA sequence of interest and a transcriptional terminator functional in a plant cell plastid, 
 wherein said plant cell is a solanaceous plant cell other than a tobacco cell.  
 
     
     
         22 . A plant cell according to  claim 11 , wherein said construct comprises regions of homology to the genome of said plastid whereby said construct is integrated into the plant cell plastid genome.  
     
     
         23 . The plant cell according to  claim 12 , wherein said plastid homology regions are derived from a heterologous plastid genome.  
     
     
         24 . The plant cell according to  claim 12 , wherein said plastid homology regions are derived from a homologous plastid genome.  
     
     
         25 . The plant cell according to  claim 11  wherein said plant cell is a potato cell.  
     
     
         26 . The plant cell according to  claim 16  wherein said plant cell comprises a leaf cell.  
     
     
         27 . The plant cell according to  claim 16  wherein said plant cell comprises a tuber cell.  
     
     
         28 . The plant cell according to  claim 11  wherein said plant cell is a petunia cell.  
     
     
         29 . The plant cell according to  claim 19  wherein said plant cell comprises a leaf cell.  
     
     
         30 . A plant, plant seed, plant cell or progeny thereof containing a plant cell according to  claim 15.