Plant promoters and uses thereof

Abstract

The invention concerns tools, methods and compositions for modifying plants and/or protein expression in plants. The invention concerns in particular transcriptional promoters enabling specific expression in the trichomes, constructs containing said promoters, and their uses for genetically modifying cells, seeds or plants. The invention also concerns methods for producing transgenic plants expressing proteins or metabolites of interest. The invention is generally applicable to any plant having glandular trichomes, and to the expression of any protein of industrial interest, in particular therapeutic or phytosanitary.

We claim: 
     
         1 . A composition of matter comprising:
 a) an isolated nucleic acid comprising:
 (i) SEQ ID NO: 2; 
 (ii) a fragment of SEQ ID NO: 2 comprising at least 100 consecutive nucleotides and having functional transcriptional promoter activity in glandular trichomes; 
 (iii) a nucleic acid sequence having at least 80% identity to SEQ ID NO: 2 or a fragment of SEQ ID NO: 2 comprising at least 100 consecutive nucleotides, said nucleic acid sequence having functional transcriptional promoter activity in glandular trichomes; 
 (iv) a sequence displaying at least 90% identity with a sequence of SEQ ID NO: 2, or a fragment thereof having at least 100 consecutive nucleotides and having functional transcriptional promoter activity in glandular trichomes; 
 (v) a nucleic acid sequence selected from SEQ ID NOs: 3 or 4 or a fragment of SEQ ID NOs: 3 or 4 comprising at least 100 consecutive nucleotides and having functional transcriptional promoter activity in glandular trichomes; 
 (vi) a nucleic acid sequence selected from SEQ ID NO: 6, 7 or 8; or 
 (vii) a nucleic acid encoding a plant cell chloroplast transit peptide comprising SEQ ID NO: 10; 
   b) a recombinant expression cassette comprising a gene of interest operatively linked to a nucleic acid comprising:
 (i) SEQ ID NO: 2; 
 (ii) a fragment of SEQ ID NO: 2 comprising at least 100 consecutive nucleotides and having functional transcriptional promoter activity in glandular trichomes; 
 (iii) a nucleic acid sequence having at least 80% identity to SEQ ID NO: 2 or a fragment of SEQ ID NO: 2 comprising at least 100 consecutive nucleotides, said nucleic acid sequence having functional transcriptional promoter activity in glandular trichomes; 
 (iv) a sequence displaying at least 90% identity with a sequence of SEQ ID NO: 2, or a fragment thereof having at least 100 consecutive nucleotides and having functional transcriptional promoter activity in glandular trichomes; 
 (v) a nucleic acid sequence selected from SEQ ID NOs: 3 or 4 or a fragment of SEQ ID NOs: 3 or 4 comprising at least 100 consecutive nucleotides and having functional transcriptional promoter activity in glandular trichomes; or 
 (vi) a nucleic acid sequence selected from SEQ ID NOs: 6, 7 or 8; 
   c) an expression vector comprising a gene of interest operatively linked to a nucleic acid comprising:
 (i) SEQ ID NO: 2; 
 (ii) a fragment of SEQ ID NO: 2 comprising at least 100 consecutive nucleotides and having functional transcriptional promoter activity in glandular trichomes; 
 (iii) a nucleic acid sequence having at least 80% identity to SEQ ID NO: 2 or a fragment of SEQ ID NO: 2 comprising at least 100 consecutive nucleotides, said nucleic acid sequence having functional transcriptional promoter activity in glandular trichomes; 
 (iv) a sequence displaying at least 90% identity with a sequence of SEQ ID NO: 2, or a fragment thereof having at least 100 consecutive nucleotides and having functional transcriptional promoter activity in glandular trichomes; 
 (v) a nucleic acid sequence selected from SEQ ID NOs: 3 or 4 or a fragment of SEQ ID NOs: 3 or 4 comprising at least 100 consecutive nucleotides and having functional transcriptional promoter activity in glandular trichomes; or 
 (vi) a nucleic acid sequence selected from SEQ ID NOs: 6, 7 or 8; 
   d) a recombinant cell comprising a recombinant expression cassette, said recombinant expression cassette comprising a gene of interest operatively linked to a nucleic acid comprising:
 (i) SEQ ID No: 2; 
 (ii) a fragment of SEQ ID NO: 2 comprising at least 100 consecutive nucleotides and having functional transcriptional promoter activity in glandular trichomes; 
 (iii) a nucleic acid sequence having at least 80% identity to SEQ ID NO: 2 or a fragment of SEQ ID NO: 2 comprising at least 100 consecutive nucleotides, said nucleic acid sequence having functional transcriptional promoter activity in glandular trichomes; 
 (iv) a sequence displaying at least 90% identity with a sequence of SEQ ID NO: 2, or a fragment thereof having at least 100 consecutive nucleotides and having functional transcriptional promoter activity in glandular trichomes; 
 (v) a nucleic acid sequence selected from SEQ ID NOs: 3 or 4 or a fragment of SEQ ID NOs: 3 or 4 comprising at least 100 consecutive nucleotides and having functional transcriptional promoter activity in glandular trichomes; or 
 (vi) a nucleic acid sequence selected from SEQ ID NOs. 6, 7 or 8; 
   e) a transgenic plant or transgenic seed comprising a recombinant expression cassette, said recombinant expression cassette comprising a gene of interest operatively linked to a nucleic acid comprising:
 (i) SEQ ID NO: 2; 
 (ii) a fragment of SEQ ID NO: 2 comprising at least 100 consecutive nucleotides and having functional transcriptional promoter activity in glandular trichomes; 
 (iii) a nucleic acid sequence having at least 80% identity to SEQ ID NO: 2 or a fragment of SEQ ID NO: 2 comprising at least 100 consecutive nucleotides, said nucleic acid sequence having functional transcriptional promoter activity in glandular trichomes; 
 (iv) a sequence displaying at least 90% identity with a sequence of SEQ ID NO: 2, or a fragment thereof having at least 100 consecutive nucleotides and having functional transcriptional promoter activity in glandular trichomes; 
 (v) a nucleic acid sequence selected from SEQ ID NOs: 3 or 4 or a fragment of SEQ ID NOs: 3 or 4 comprising at least 100 consecutive nucleotides and having functional transcriptional promoter activity in glandular trichomes; or 
 (vi) a nucleic acid sequence selected from SEQ ID NOs: 6, 7 or 8; or 
   f) an isolated plant cell chloroplast transit peptide comprising SEQ ID NO: 10.   
     
     
         2 . The composition of matter according to  claim 1 , wherein said expression cassette comprises a gene of interest that encodes an enzyme leading to the modification of the composition of the exudate secreted by the trichomes. 
     
     
         3 . The composition of matter according to  claim 1 , wherein said expression cassette comprises a gene of interest that encodes a therapeutic or phytosanitary protein, an enzyme, a resistance protein, a transcriptional activator or a viral genome. 
     
     
         4 . The composition of matter according to  claim 1 , wherein said expression cassette further comprises a nucleic acid encoding SEQ ID NO: 10. 
     
     
         5 . The composition of matter according to  claim 1 , wherein said recombinant cell is a plant cell. 
     
     
         6 . The composition of matter according to  claim 5 , wherein said plant cell is from the family Solanaceae, Asteraceae, Cannabaceae or Lamiaceae. 
     
     
         7 . A method for producing a protein in a plant comprising introducing into a plant cell or seed a recombinant cassette comprising a gene of interest that encodes a protein of interest and regenerating a plant from said plant cell or seed that produces said protein of interest, said recombinant expression cassette comprising a gene of interest operatively linked to a nucleic acid comprising:
 (i) SEQ ID NO: 2;   (ii) a fragment of SEQ ID NO: 2 comprising at least 100 consecutive nucleotides and having functional transcriptional promoter activity in glandular trichomes;   (iii) a nucleic acid sequence having at least 80% identity to SEQ ID NO: 2 or a fragment of SEQ ID NO: 2 comprising at least 100 consecutive nucleotides, said nucleic acid sequence having functional transcriptional promoter activity in glandular trichomes;   (iv) a sequence displaying at least 90% identity with a sequence of SEQ ID NO: 2, or a fragment thereof having at least 100 consecutive nucleotides and having functional transcriptional promoter activity in glandular trichomes;   (v) a nucleic acid sequence selected from SEQ ID NOs: 3 or 4 or a fragment of SEQ ID NOs: 3 or 4 comprising at least 100 consecutive nucleotides and having functional transcriptional promoter activity in glandular trichomes; or   (vi) a nucleic acid sequence selected from SEQ ID NOs: 6, 7 or 8.   
     
     
         8 . The method according to  claim 7 , further comprising recovering the protein of interest from exudate at the leaf surface. 
     
     
         9 . The method according to  claim 7 , wherein the plant, plant seed or plant cell is from the family Solanaceae, Asteraceae, Cannabaceae or Lamiaceae.