Chimeric promoters capable of mediating gene expression in plants upon pathogen infection and uses thereof

Patent metadata

Field	Value
Publication number	US-9297018-B2
Application number	US-201313930266-A
Country	US
Kind code	B2
Filing date	Jun 28, 2013
Priority date	Nov 12, 1998
Publication date	Mar 29, 2016
Grant date	Mar 29, 2016

Abstract

Described are synthetic promoters capable of mediating gene expression in plants upon pathogen infection. Furthermore, recombinant genes and vectors comprising said chimeric promoters as well as host cells transformed with such chimeric promoters, recombinant genes, or vectors are provided. Additionally, diagnostic compositions and kits comprising such chimeric promoters, recombinant genes, vectors or cells are described. Provided are further methods for the identification of compounds being capable of activating or inhibiting genes that are specifically expressed in plants upon pathogen infection employing the above described means. Furthermore, transgenic plant cells, plant tissue, and plants containing the above-described chimeric promoters, recombinant genes, and vectors as well as the use of the aforementioned chimeric promoters, recombinant genes, vectors and/or compounds identified by the method of the invention in plant cell and tissue culture, plant breeding, and/or agriculture are described.

First claim

What is claimed is:

1. A chimeric promoter capable of local gene expression in plants of an operably linked nucleic acid sequence, wherein the expression is induced by a pathogen elicitor treatment, a pathogen infection, or both, wherein the chimeric promoter comprises:
(i) at least one cis-acting element sufficient to direct: pathogen-elicitor-specific expression of the nucleic acid sequence, pathogen-infection-specific expression of the nucleic acid sequence, or both, wherein the at least one cis-acting element comprises the nucleotide sequence of SEQ ID NO: 7 or 14, and
(ii) a minimal promoter.

2. The chimeric promoter according to claim 1 , further comprising at least one additional cis-acting element consisting of the nucleotide sequence selected from the group consisting of the nucleotide sequences of one of SEQ ID NO: 1-16.

3. The chimeric promoter of claim 2 , wherein said chimeric promoter comprises homo- and/or hetero-multimeric forms of said at least one cis-acting element and said at least one additional cis-acting element.

4. The chimeric promoter of claim 3 , wherein said homo- and/or hetero-multimeric form is a dimer or a tetramer.

5. The chimeric promoter of claim 2 wherein at least two of the elements selected from said at least one cis-acting element and said at least one additional cis-acting element are separated by a spacer, wherein the spacer is (a) from 4 to 10 or (b) about 50 to about 1000 base pairs.

6. The chimeric promoter of claim 2 wherein at least one of said at least one cis-acting element and one of said at least one additional cis-acting element are separated by a spacer, wherein the spacer is (a) from 4 to 10 or (b) about 50 to about 1000 base pairs.

7. The chimeric promoter according to claim 1 , wherein the at least one cis-acting element comprises two copies of the nucleotide sequence of SEQ ID NO: 7 or 14.

8. The chimeric promoter according to claim 7 , further comprising two copies of the nucleotide sequences of SEQ ID NO:3 or 4.

9. The chimeric promoter of claim 1 , wherein the minimal promoter is selected from the group consisting of a CaMV35S promoter, a CHS promoter, a PR1 promoter, and a hcbt2 promoter.

10. The chimeric promoter of claim 1 , wherein the distance between said at least one cis-acting element and said minimal promoter is 12 to 300 base pairs.

11. The chimeric promoter of claim 1 , wherein the distance between said at least one cis-acting element and said minimal promoter is 25 to 70 base pairs.

12. The chimeric promoter of claim 1 , wherein the distance between said at least one cis-acting element and said minimal promoter is 40 to 60 base pairs.

13. A recombinant gene comprising the chimeric promoter of claim 1 .

14. The recombinant gene of claim 13 , wherein at least one of the at least one cis-acting elements is located in the 5′- or 3′-untranslated region or in an intron of the recombinant gene.

15. The recombinant gene of claim 13 , wherein the chimeric promoter is operatively linked to a heterologous DNA sequence, and wherein said heterologous DNA sequence encodes a (poly)peptide, a cytotoxic protein, an antibody, an antisense RNA, a sense RNA, a ribozyme, a transcription factor, a protease, a nuclease, a lipase, or a polymerase.

16. A vector comprising the chimeric promoter of claim 1 or the recombinant gene of claim 13 .

17. A transgenic plant cell comprising the chimeric promoter of claim 1 , the recombinant gene of claim 13 , or a vector comprising the chimeric promoter of claim 1 or the recombinant gene of claim 13 .

18. A transgenic plant or a plant tissue comprising the transgenic plant cell of claim 17 .

19. A harvestable part of a transgenic plant comprising the transgenic plant cell of claim 17 .

20. A propagation material of a transgenic plant comprising the transgenic plant cell of claim 17 .

21. An isolated cis-acting element sufficient to direct pathogen-elicitor-specific expression, pathogen-infection-specific expression, or both, consisting of the nucleotide sequence of SEQ ID NO: 7 or 14.

22. A chimeric promoter obtained by a method of rendering a promoter of a gene responsive to pathogens comprising inserting at least one cis-acting element sufficient to direct pathogen-elicitor-specific expression, pathogen-infection-specific expression, or both, into the promoter of said gene, wherein the at least one cis-acting element comprises the nucleotide sequences of SEQ ID NO: 7 or 14 to produce the chimeric promoter.

23. An isolated chimeric promoter to render a gene responsive to pathogens, obtained by a method comprising
inserting at least one cis-acting element sufficient to direct pathogen-elicitor-induced expression, pathogen-infection induced expression, or both, of an operably linked nucleic acid, into the promoter of said gene, wherein the at least one cis-acting element comprises the nucleotide sequence of SEQ ID NO: 7 or 14.

24. A recombinant gene wherein the isolated chimeric promoter of claim 23 is linked to a coding sequence of a gene.

25. A vector wherein the isolated chimeric promoter of claim 23 is linked to vector sequences.

26. A chimeric promoter obtained by a method of rendering a promoter of a gene responsive to pathogens comprising inserting at least one cis-acting element sufficient to direct pathogen-elicitor-specific expression, pathogen-infection-specific expression, or both, into the promoter of said gene, wherein the at least one cis-acting element comprises two copies of the nucleotide sequences of SEQ ID NO: 7 or 14, or a combination of one copy of the nucleotide sequence of SEQ ID NO: 7 or 14 and one copy selected from the group consisting of one of the nucleotide sequences of SEQ ID NO: 1-16 to produce the chimeric promoter.

Assignees

• Max Planck Gesellschaft

Inventors

• Kirsch Christoph
• Logemann Elke
• Hahlbrock Klaus
• Rushton Paul
• Somssich Imre

Classifications

• C07K14/415 Primary

  from plants · CPC title

• C07K16/16

  against material from plants · CPC title

• C12N15/8239

  pathogen inducible · CPC title

• C12N15/8216 Primary

  Methods for controlling, regulating or enhancing expression of transgenes in plant cells · CPC title

• C12Q1/025

  for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics (antimicrobial activity C12Q1/18) · CPC title