Pathogen-resistant plants transformed with a DNA encoding sarcotoxin 1A linked to a signal peptide and a method for production thereof

• US 6,750,381 B2
• Filed: 02/26/1997
• Issued: 06/15/2004
• Est. Priority Date: 03/25/1996
• Status: Expired due to Fees

• Current Assignee: National Institute of Agrobiological Sciences
• Original Assignee: National Institute of Agrobiological Sciences
• Inventors: Ohashi, Yuko, Ohshima, Masahiro, Mitsuhara, Ichiro
• Primary Examiner(s): Nelson, Amy J.
• Assistant Examiner(s): KUBELIK, ANNE R
• Application Number:
  US08/805,813
• Publication Number:
  US 20020166142A1
• Time in Patent Office: 2,666 Days
• Field of Search: 800/279, 800/301, 800/205, 536/23.6, 536/23.7, 435/172.3, 435/320.1, 435/419
• US Class Current: 800/301
• CPC Class Codes: C07K 14/43577 from flies
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C12N 15/8281 for bacterial resistance
• CPC Class Codes: C12N 15/8282 for fungal resistance

Abstract

The present invention is drawn to a method of conferring resistance to pathogenic fungi on a plant by transformation with a nucleic acid encoding Sarcotoxin 1a operatively linked to a signal peptide, and optionally linked via the hinge region of tobacco chitinase to a plant peptide, and plants thereby obtained.

Claims

1. A method of conferring resistance to pathogenic fungi on a plant, wherein the method comprises the steps of:

transforming a plant cell with an expression vector, wherein said expression vector comprises;

an expression cassette comprising a first plant promoter induced by stress operably linked to a DNA sequence encoding Sarcotoxin 1a, wherein a DNA sequence encoding a signal peptide is operatively positioned between the first plant promoter and the DNA sequence encoding Sarcotoxin 1a; and

a constitutively expressed plant promoter positioned adjacent to the first plant promoter; and

regenerating the plant cell into a transgenic plant, wherein the transgenic plant has enhanced resistance to pathogenic fungi as compared to a corresponding untransformed plant.

2. The method according to claim 1, wherein said expression vector further comprises a drug resistance gene operably linked to the constitutively expressed plant promoter.

3. The method according to claim 1, wherein a DNA sequence encoding a plant protein is operatively fused to the DNA sequence encoding Sarcotoxin 1a via the hinge region of the tobacco chitinase gene.

4. The method according to claim 1, wherein the DNA sequence encoding the signal peptide is from a plant gene.

5. The method according to claim 1, wherein the promoter induced by stress is the promoter of a tobacco PR-1a gene.

6. The method according to claim 2, wherein the expression cassette further comprises the terminator of the tobacco PR-1a gene operably linked downstream of the DNA sequence encoding Sarcotoxin 1a.

7. The method according to claim 1, wherein the constitutively expressed plant promoter is a cauliflower mosaic virus 35S promoter.

8. The method according to claim 1, wherein the signal peptide is the PR-1a signal peptide.

9. A transgenic plant which is resistant to pathogenic fungi, wherein the plant comprises an expression vector, wherein the expression vector comprises:

a first expression cassette comprising a DNA sequence encoding Sarcotoxin 1a operably linked to a promoter induced by stress, wherein a DNA sequence encoding a signal peptide is operatively positioned between the promoter induced by stress and the DNA sequence encoding Sarcotoxin 1a; and

a second expression cassette comprising a drug resistance gene operably linked to a constitutively expressed promoter;

wherein the first and second expression cassettes are positioned adjacent to each other, and wherein the transgenic plant has enhanced resistance to pathogenic fungi as compared to a corresponding untransformed plant.

10. The plant according to claim 9, wherein a DNA sequence encoding a plant protein is operatively fused to the DNA sequence encoding Sarcotoxin 1a via the hinge region of the tobacco chitinase gene.

11. The plant according to claim 9, wherein the DNA sequence encoding the signal peptide is from a plant gene.

12. The plant according to claim 9, wherein the promoter induced by stress is a promoter of the tobacco PR-1a gene.

13. The plant according to claim 9, wherein the first expression cassette further comprises the terminator of the tobacco PR-1a gene operably linked downstream of the DNA sequence encoding Sarcotoxin 1a.

14. The plant according to claim 9, wherein the constitutively expressed promoter is a cauliflower mosaic virus 35 S promoter.

15. The plant according to claim 9, wherein the expression vector further comprises a T-DNA region.

16. The plant according to claim 9, wherein the signal peptide is the PR-1a signal peptide.