Transgenic plants expressing DNA constructs containing a plurality of genes to impart virus resistance

• US 6,337,431 B1
• Filed: 10/06/1997
• Issued: 01/08/2002
• Est. Priority Date: 12/30/1994
• Status: Expired due to Term

• Current Assignee: Seminis Vegetable Seeds Incorporated (Bayer AG)
• Original Assignee: Seminis Vegetable Seeds Incorporated (Bayer AG)
• Inventors: Russell, Paul F., Reynolds, John F., Carney, Kim J., Tricoli, David M, Deng, Rosaline Z., Quemada, Hector D., McMaster, Russell J.
• Primary Examiner(s): McElwain, Elizabeth F.
• Assistant Examiner(s): MEHTA, ASHWIN D
• Application Number:
  US08/860,379
• Time in Patent Office: 1,555 Days
• Field of Search: 800/205, 800/278, 800/279, 800/280, 800/288, 800/295, 800/298, 800/301, 800/317, 435/172.3, 435/320.1, 435/69.1, 435/410, 435/419, 435/468, 536/23.72
• US Class Current: 800/280
• CPC Class Codes: C07K 14/005 from viruses
• CPC Class Codes: C12N 15/8247 involving modified lipid metabolism, e.g. seed oil composition
• CPC Class Codes: C12N 15/8283 for virus resistance
• CPC Class Codes: C12N 2770/00022 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: C12N 2770/34022 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes

Abstract

The present invention provides a chimeric recombinant DNA molecule comprising: a plurality of DNA sequences, each of which comprises a plant-functional promoter linked to a coding region, which encodes a virus-associated coat protein, wherein said DNA sequences are preferably linked in tandem so that they are expressed in virus-susceptible plant cells transformed with said recombinant DNA molecule to impart resistance to said viruses; as well as methods for transforming plants with the chimeric recombinant DNA molecule and for selecting plants which express at least one of said DNA sequences imparting viral resistance.

Claims

1. A chimeric recombinant DNA molecule comprising:

at least three tandemly linked DNA sequences, wherein each DNA sequence comprises a promoter operably linked to a DNA sequence which encodes a viral coat protein, wherein said tandemly linked DNA sequences impart substantially the same level of resistance against viral disease to plant cells caused by each of the viruses from which the tandemly linked DNA sequences originated, and further wherein said viruses are chosen from the group consisting of potyviruses, cucomoviruses, and comoviruses.

2. The DNA molecule of claim 1 wherein said DNA sequences further comprise a selectable marker gene, a reporter gene or a combination thereof that enables identification of plant cells transformed with said DNA molecule.

3. The DNA molecule of claim 2, wherein the DNA sequences are flanked by two selectable marker genes, two reporter genes or a combination thereof.

4. The DNA molecule of claim 1 wherein said viral coat proteins comprise the coat proteins of watermelon mosaic virus II, cucumber mosaic virus, and zucchini yellow mosaic virus.

5. The DNA molecule of claim 1 wherein the plant cells are cells derived from a member of the Cucurbitaceae family.

6. A method of imparting multi-virus resistance to a plant which is susceptible to viruses, comprising:

(a) transforming cells of said susceptible plant with a recombinant DNA molecule comprising at least three tandemly linked DNA sequences, each comprising a promoter functional in cells of said plant and operably linked to a DNA sequence encoding a coat protein of a virus which is capable of infecting said plant, wherein said viruses are chosen from the group of viruses consisting of potyviruses, cucomoviruses, and comoviruses;

(b) regenerating said plant cells to provide a differentiated plant; and

(c) identifying a transformed plant which contains the tandemly linked DNA sequences which renders substantially equal levels of resistance against viral disease caused by said viruses to the plant.

7. The method of claim 6, wherein the expression of at least one of said DNA sequences imparts resistance to more than one virus.

8. The method of claim 6, wherein the plant is a dicot.

9. The method of claim 6, wherein the DNA molecule is part of a binary Ti plasmid and the plant cells are transformed by A. tumefaciens mediated transformation.

10. The method of claim 6, wherein the DNA sequences further comprise a selectable marker gene or a reporter gene that enables identification of said transformed plant.

11. The method of claim 6, wherein said DNA sequences further comprise the coat protein genes of watermelon mosaic virus II, cucumber mosaic virus and zucchini yellow mosaic virus.

12. The method of claim 6, wherein the susceptible plant is a member of the Cucurbitaceae family.

13. A transformed plant prepared by the method of claim 6.

14. A transformed plant cell prepared by the method of claim 6.

15. A transformed seed of the transformed plant of claim 13.

16. A hybrid plant, prepared by crossing the plant of claim 13, with a second plant, which is resistant to viral disease caused by said viruses.