Expression cassette encoding a 5-enolpyruvylshikimate-3-phosphate synthase (epsps) and herbicde-tolerant plants containing it

Abstract

The present invention relates to a novel expression cassette comprising a nucleic acid sequence encoding an EPSPS. In particular, the present invention relates to a novel expression cassette comprising, in the direction of transcription, functionally linked to one another, a promoter regulatory sequence which is functional in plant cells or plants, a nucleic acid sequence encoding an EPSPS and a terminator sequence which is functional in plant cells or plants, characterized in that the promoter regulatory sequence is a nucleic acid sequence chosen from the promoter regulatory sequences of the CsVMV (Cassaya Vein Mosaic Virus) plant virus.

1 . An expression cassette comprising, in the direction of transcription, functionally linked to one another, a promoter regulatory sequence which is functional in plant cells or plants, a nucleic acid sequence encoding an EPSPS and a terminator regulatory sequence which is functional in plant cells or plants, characterized in that the promoter regulatory sequence is a nucleic acid sequence chosen from the promoter regulatory sequences of the CsVMV (Cassaya Vein Mosaic Virus) plant virus.  
     
     
         2 . The expression cassette as claimed in  claim 1 , characterized in that the promoter regulatory sequence comprises, in the direction of transcription, the nucleic acid sequences X, Y and Z as defined respectively by SEQ ID NOS 1, 2 and 3.  
     
     
         3 . The expression cassette as claimed in  claim 1 , characterized in that the promoter regulatory sequence is represented by SEQ ID NO 4.  
     
     
         4 . The expression cassette as claimed in  claim 1 , characterized in that the promoter regulatory sequence is represented by SEQ ID NO 5.  
     
     
         5 . The expression cassette as claimed in  claim 1 , characterized in that the nucleic acid sequence encoding a EPSPS is a sequence encoding an EPSPS which has been mutated at the amino acids corresponding to the threonine at position 102 and to the proline at position 106, said positions being relative with respect to the maize EPSPS sequence.  
     
     
         6 . The expression cassette as claimed in  claim 5 , characterized in that the nucleic acid sequence encoding an EPSPS is the sequence encoding a mutated EPSPS comprising an isoleucine at position 102 and a serine at position 106, said positions being relative with respect to the maize EPSPS sequence.  
     
     
         7 . The expression cassette as claimed in  claim 6 , characterized in that the nucleic acid sequence encoding a mutated EPSPS is the sequence encoding a maize mutated EPSPS comprising an isoleucine at position 102 and a serine at position 106.  
     
     
         8 . The expression cassette as claimed in  claim 1 , characterized in that it also comprises a double transit peptide comprising, in the direction of transcription, a sequence encoding a transit peptide of a plant gene encoding an enzyme which is located in plastids, a portion of sequence of the mature N-terminal portion of a plant gene encoding an enzyme which is located in plastids, and then a sequence encoding a second transit peptide of a plant gene encoding an enzyme which is located in plastids.  
     
     
         9 . A cloning and/or expression vector for transforming plant cells or plants, characterized in that it comprises at least one expression cassette as claimed in  claim 1 .  
     
     
         10 . A transformed plant cell, characterized in that it comprises an expression cassette as claimed in  claim 1 .  
     
     
         11 . A transformed plant, characterized in that it comprises an expression cassette as claimed in  claim 1 .  
     
     
         12 . A seed of a transformed plant as claimed in  claim 11 , characterized in that it comprises an expression cassette comprising, in the direction of transcription, functionally linked to one another, a promoter regulatory sequence which is functional in plant cells or plants, a nucleic acid sequence encoding an EPSPS and a terminator regulatory sequence which is functional in plant cells or plants, characterized in that the promoter regulatory sequence is a nucleic acid sequence chosen from the promoter regulatory sequences of the CsVMV (Cassaya Vein Mosaic Virus) plant virus, said expression cassette also comprising a double transit peptide comprising, in the direction of transcription, a sequence encoding a transit peptide of a plant gene encoding an enzyme which is located in plastids, a portion of sequence of the mature N-terminal portion of a plant gene encoding an enzyme which is located in plastids, and then a sequence encoding a second transit peptide of a plant gene encoding an enzyme which is located in plastids.  
     
     
         13 . A method for treating plants as claimed in  claim 11 , characterized in that said plants are treated with EPSPS-inhibiting herbicide.  
     
     
         14 . The method as claimed in  claim 13 , characterized in that said EPSPS-inhibiting herbicide is glyphosate.  
     
     
         15 . A method for controlling weeds in crops, characterized in that transformed plants as claimed in  claim 11  are grown, and in that said plants are treated with an EPSPS-inhibiting herbicide.  
     
     
         16 . The method as claimed in  claim 15 , characterized in that said EPSPS-inhibiting herbicide is glyphosate.  
     
     
         17 . A method for growing transformed plants as claimed in  claim 11 , characterized in that it consists in planting seeds of said transformed plants in an area of a field suitable for growing said plants, in applying to said area of said field an agrochemical composition, without substantially affecting said transformed seeds or said transformed plants, then in harvesting the plants grown when they have reached the desired maturity and, optionally, in separating the seeds from the harvested plants.