DNA comprising plum pox virus and tomato spotted wilt virus cDNAS for disease resistance

• US 5,569,823 A
• Filed: 05/23/1994
• Issued: 10/29/1996
• Est. Priority Date: 05/28/1993
• Status: Expired due to Fees

• Current Assignee: Bayer AG
• Original Assignee: Bayer AG
• Inventors: Stenzel, Klaus, Maiss, Edgar, Adam, G unter, Schreier, Peter H.
• Primary Examiner(s): Robinson, Douglas W.
• Assistant Examiner(s): McElwain, Elizabeth F.
• Application Number:
  US08/247,809
• Time in Patent Office: 890 Days
• Field of Search: 800/205, 536/23.1, 536/23.72, 435/172.3, 435/240.4, 435/320.1, 935/30, 935/67
• US Class Current: 800/280
• CPC Class Codes: C07K 14/005 from viruses
• CPC Class Codes: C12N 15/8283 for virus resistance
• CPC Class Codes: C12N 15/8285 for nematode resistance
• CPC Class Codes: C12N 2770/34022 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: Y02A 40/146 Genetically Modified [GMO] plants, e.g. transgenic plants

Abstract

The present invention relates to novel, recombinant deoxyribonucleic acids (DNA), to vectors and host organisms which contain them, and to transgenic plants which contain the recombinant DNA and possess an elevated resistance toward pernicious organisms and plant diseases, where the recombinant deoxyribonucleic acids (DNA) are characterized in that they consist of a combination of the following components, or components possessing a DNA which in each case has the same effect, or contain these constituents:

(a) a double-stranded cDNA fragment which is derived from the RNA of the plum pox virus (PPV) ("fragment A"), and

(b) a double-stranded cDNA fragment which is derived from the S RNA of the tomato spotted wilt virus (TSWV) ("fragment B"), it being possible for these combinations also to possess, in addition to further DNA fragments which may optionally be present, a promoter which is active in plant cells.

Claims

1. DNA comprising a combination of fragment A linked directly or indirectly to fragment B, wherein:

fragment A consists of a double-stranded cDNA fragment derived from the RNA of the plum pox virus; and

fragment B consists of a double-stranded cDNA fragment derived from the S RNA of the N structural gene of the tomato spotted wilt virus;

said combination, when expressed in a plant, conferring on said plant an increased resistance, as compared to a natural plant of the same species, to tomato spotted wilt virus and another tospovirus.

2. DNA according to claim 1, wherein fragment B is situated after fragment A in the 3'"'"' direction.

3. DNA according to claim 1, wherein fragment A comprises SEQ ID NO:

7.

4. DNA according to claim 3, wherein fragment A consists of SEQ ID NO:

7.

5. DNA according to claim 1, wherein fragment B comprises SEQ ID NO:

11.

6. DNA according to claim 5, wherein fragment B consists of SEQ ID NO:

11.

7. DNA according to claim 5, wherein fragment A comprises SEQ ID NO:

7.

8. DNA according to claim 7, wherein fragment A consists of SEQ ID NO:

7.

9. DNA according to claim 1, further comprising a promoter which is active in a plant.

10. DNA according to claim 9, wherein the promoter is the CaMV 35S promoter.

11. DNA according to claim 1, further comprising fragment C, which is a synthetic DNA sequence having 1 to 134 base pairs and which connects the open reading frame in fragment A to fragment B.

12. DNA according to claim 11, wherein fragment C comprises SEQ ID NO:

9.

13. DNA according to claim 12, wherein fragment C consists of SEQ ID NO:

9.

14. DNA according to claim 1, further comprising fragment D, which is a double-stranded DNA sequence having 1 to 30 base pairs and follows fragment B in the 3'"'"' direction.

15. DNA according to claim 14, wherein fragment D comprises SEQ ID NO:

13.

16. DNA according to claim 15, wherein fragment D consists of SEQ ID NO:

13.

17. DNA according to claim 1, further comprising a 3'"'"' untranslated DNA which follows fragment B directly in the 3'"'"' direction or is connected to fragment B through fragment D, which is a double-stranded DNA sequence having 1 to 30 base pairs.

18. DNA according to claim 17, wherein the 3'"'"' untranslated DNA comprises SEQ ID NO:

4.

19. DNA according to claim 18 wherein the 3'"'"' untranslated DNA consists of SEQ ID NO:

4.

27. A plasmid or phage vector containing DNA according to claim 1, 20 or 22.

28. A transformed plant cell, said plant cell containing in its genome DNA in addition to the DNA naturally constituting the genome of said plant cell, said plant cell expressing said additional DNA, said additional DNA comprising DNA according to claim 1, and said transformed plant cell, as a result of the expression of said additional DNA, exhibiting an increased resistance, as compared to a plant cell of the same plant species and cell type that is not so transformed, to tomato spotted wilt virus and another tospovirus, or propagation material of said transformed plant cell.

33. A transformed whole plant, said whole plant containing in its genome DNA in addition to the DNA naturally constituting the genome of said whole plant, said whole plant expressing said additional DNA, said additional DNA comprising DNA according to claim 1, and said transformed whole plant, as a result of the expression of said additional DNA, exhibiting an increased resistance, as compared to a whole plant of the same plant species that is not so transformed, to tomato spotted wilt virus and another tospovirus, or propagation material of said transformed whole plant.

38. A transformed plant part, said plant part containing in its genome DNA in addition to the DNA naturally constituting the genome of said plant part, said plant part expressing said additional DNA, said additional DNA comprising DNA according to claim 1, and said transformed plant part, as a result of the expression of said additional DNA, exhibiting an increased resistance, as compared to a plant part of the same plant species and part type that is not so transformed, to tomato spotted wilt virus and another tospovirus, or propagation material of said transformed plant part.

51. A method of protecting a plant against infection by tomato spotted wilt virus and another tospovirus, said method comprising transforming said plant with DNA according to claim 1 in a manner such that said DNA is expressed in an amount effective to protect said plant against said infection.

20. DNA comprising SEQ ID NO:

1 or a DNA that hybridizes to SEQ ID NO;

1 under stringent hybridization conditions, said DNA that hybridizes to SEQ ID NO;

1, when expressed in a plant, conferring on said plant an increased resistance, as compared to a natural plant of the same species, to tomato spotted wilt virus and another tospovirus.

21. DNA according to claim 20, consisting of SEQ ID NO:

1 or a DNA that hybridizes to SEQ ID NO;

1 under stringent hybridization conditions, said DNA that hybridizes to SEQ ID NO;

1, when expressed in a plant, conferring on said plant an increased resistance, as compared to a natural plant of the same species, to tomato spotted wilt virus and another tospovirus.

29. A transformed plant cell, said plant cell containing in its genome DNA in addition to the DNA naturally constituting the genome of said plant cell, said plant cell expressing said additional DNA, said additional DNA comprising DNA according to claim 20, and said transformed plant cell, as a result of the expression of said additional DNA, exhibiting an increased resistance, as compared to a plant cell of the same plant species and cell type that is not so transformed, to tomato spotted wilt virus and another tospovirus, or propagation material of said transformed plant cell.

34. A transformed whole plant, said whole plant containing in its genome DNA in addition to the DNA naturally constituting the genome of said whole plant, said whole plant expressing said additional DNA, said additional DNA comprising DNA according to claim 20, and said transformed whole plant, as a result of the expression of said additional DNA, exhibiting an increased resistance, as compared to a whole plant of the same plant species that is not so transformed, to tomato spotted wilt virus and another tospovirus, or propagation material of said transformed whole plant.

39. A transformed plant part, said plant part containing in its genome DNA in addition to the DNA naturally constituting the genome of said plant part, said plant part expressing said additional DNA, said additional DNA comprising DNA according to claim 20, and said transformed plant part, as a result of the expression of said additional DNA, exhibiting an increased resistance, as compared to a plant part of the same plant species and part type that is not so transformed, to tomato spotted wilt virus and another tospovirus, or propagation material of said transformed plant part.

43. A method of protecting a plant against infection by tomato spotted wilt virus and another tospovirus, said method comprising transforming said plant with DNA according to claim 20 in a manner such that said DNA is expressed in an amount effective to protect said plant against said infection.

45. A transformed plant cell according to claim 29, wherein said additional DNA consists of SEQ ID NO:

1 or a DNA that hybridizes to SEQ ID NO;

1 under stringent hybridization conditions, said DNA that hybridizes to SEQ ID NO;

1, when expressed in said plant cell, conferring on said plant cell an increased resistance, as compared to a natural plant cell of the same species and cell type, to tomato spotted wilt virus and another tospovirus.

47. A transformed whole plant according to claim 34, wherein said additional DNA consists of SEQ ID NO:

1 or a DNA that hybridizes to SEQ ID NO;

1 under stringent hybridization conditions, said DNA that hybridizes to SEQ ID NO;

1, when expressed in said whole plant, conferring on said whole plant an increased resistance, as compared to a natural whole plant of the same species, to tomato spotted wilt virus and another tospovirus.

49. A transformed plant part according to claim 39, wherein said additional DNA consists of SEQ ID NO:

1 or a DNA that hybridizes to SEQ ID NO;

1 under stringent hybridization conditions, said DNA that hybridizes to SEQ ID NO;

1, when expressed in said plant part, conferring on said plant part an increased resistance, as compared to a natural plant part of the same species and plant part type, to tomato spotted wilt virus and another tospovirus.

52. A method according to claim 43, wherein said DNA consists of SEQ ID NO:

1 or a DNA that hybridizes to SEQ ID NO;

1 under stringent hybridization conditions, said DNA that hybridizes to SEQ ID NO;

1, when expressed in said plant, conferring on said plant an increased resistance, as compared to a natural plant of the same species, to tomato spotted wilt virus and another tospovirus.

53. A method according to claim 52, wherein said DNA consists of SEQ ID NO:

1.

22. DNA comprising SEQ ID NO:

5 or a DNA that hybridizes to SEQ ID NO;

5 under stringent hybridization conditions, said DNA that hybridizes to SEQ ID NO;

5, when expressed in a plant, conferring on said plant an increased resistance, as compared to a natural plant of the same species, to tomato spotted wilt virus and another tospovirus.

23. DNA according to claim 22, consisting of SEQ ID NO:

5 or a DNA that hybridizes to SEQ ID NO;

5 under stringent hybridization conditions, said DNA that hybridizes to SEQ ID NO;

5, when expressed in a plant, conferring on said plant an increased resistance, as compared to a natural plant of the same species, to tomato spotted wilt virus and another tospovirus.

31. A transformed plant cell, said plant cell containing in its genome DNA in addition to the DNA naturally constituting the genome of said plant cell, said plant cell expressing said additional DNA, said additional DNA comprising DNA according to claim 22, and said transformed plant cell, as a result of the expression of said additional DNA, exhibiting an increased resistance, as compared to a plant cell of the same plant species and cell type that is not so transformed, to tomato spotted wilt virus and another tospovirus, or propagation material of said transformed plant cell.

36. A transformed whole plant, said whole plant containing in its genome DNA in addition to the DNA naturally constituting the genome of said whole plant, said whole plant expressing said additional DNA, said additional DNA comprising DNA according to claim 22, and said transformed whole plant, as a result of the expression of said additional DNA, exhibiting an increased resistance, as compared to a whole plant of the same plant species that is not so transformed, to tomato spotted wilt virus and another tospovirus, or propagation material of said transformed whole plant.

41. A transformed plant part, said plant part containing in its genome DNA in addition to the DNA naturally constituting the genome of said plant part, said plant part expressing said additional DNA, said additional DNA comprising DNA according to claim 22, and said transformed plant part, as a result of the expression of said additional DNA, exhibiting an increased resistance, as compared to a plant part of the same plant species and part type that is not so transformed, to tomato spotted wilt virus and another tospovirus, or propagation material of said transformed plant part.

44. A method of protecting a plant against infection by tomato spotted wilt virus and another tospovirus, said method comprising transforming said plant with DNA according to claim 22 in a manner such that said DNA is expressed in an amount effective to protect said plant against said infection.

46. A transformed plant cell according to claim 31, wherein said additional DNA consists of SEQ ID NO:

5 or a DNA that hybridizes to SEQ ID NO;

5 under stringent hybridization conditions, said DNA that hybridizes to SEQ ID NO;

5, when expressed in said plant cell, conferring on said plant cell an increased resistance, as compared to a natural plant cell of the same species and cell type, to tomato spotted wilt virus and another tospovirus.

48. A transformed whole plant according to claim 36, wherein said additional DNA consists of SEQ ID NO:

5 or a DNA that hybridizes to SEQ ID NO;

5 under stringent hybridization conditions, said DNA that hybridizes to SEQ ID NO;

5, when expressed in said whole plant, conferring on said whole plant an increased resistance, as compared to a natural whole plant of the same species, to tomato spotted wilt virus and another tospovirus.

50. A transformed plant part according to claim 41, wherein said additional DNA consists of SEQ ID NO:

5 or a DNA that hybridizes to SEQ ID NO;

5 under stringent hybridization conditions, said DNA that hybridizes to SEQ ID NO;

5, when expressed in said plant part, conferring on said plant part an increased resistance, as compared to a natural plant part of the same species and plant part type, to tomato spotted wilt virus and another tospovirus.

54. A method according to claim 44, wherein said DNA consists of SEQ ID NO:

5 or a DNA that hybridizes to SEQ ID NO;

5 under stringent hybridization conditions, said DNA that hybridizes to SEQ ID NO;

5, when expressed in said plant, conferring on said plant an increased resistance, as compared to a natural plant of the same species, to tomato spotted wilt virus and another tospovirus.

55. A method according to claim 54, wherein said DNA consists of SEQ ID NO:

5.

24. Isolated DNA selected from the group consisting of SEQ ID NOS:

1 and 5.

25. Isolated DNA according to claim 24, which is SEQ ID NO:

1.

26. Isolated DNA according to claim 24, which is SEQ ID NO:

5.

30. A transformed plant cell, said plant cell containing in its genome DNA in addition to the DNA naturally constituting the genome of said plant cell, said plant cell expressing said additional DNA, said additional DNA consisting of SEQ ID No:

1, and said transformed plant cell, as a result of the expression of said additional DNA, exhibiting an increased resistance, as compared to a plant cell of the same plant species and cell type that is not so transformed, to tomato spotted wilt virus and another tospovirus, or propagation material of said transformed plant cell.

32. A transformed plant cell, said plant cell containing in its genome DNA in addition to the DNA naturally constituting the genome of said plant cell, said plant cell expressing said additional DNA, said additional DNA consisting of SEQ ID No:

5, and said transformed plant cell, as a result of the expression of said additional DNA, exhibiting an increased resistance, as compared to a plant cell of the same plant species and cell type that is not so transformed, to tomato spotted wilt virus and another tospovirus, or propagation material of said transformed plant cell.

35. A transformed whole plant, said whole plant containing in its genome DNA in addition to the DNA naturally constituting the genome of said whole plant, said whole plant expressing said additional DNA, said additional DNA consisting of SEQ ID No:

1, and said transformed whole plant, as a result of the expression of said additional DNA, exhibiting an increased resistance, as compared to a whole plant of the same plant species that is not so transformed, to tomato spotted wilt virus and another tospovirus, or propagation material of said transformed whole plant.

37. A transformed whole plant, said whole plant containing in its genome DNA in addition to the DNA naturally constituting the genome of said whole plant, said whole plant expressing said additional DNA, said additional DNA consisting of SEQ ID No:

5, and said transformed whole plant, as a result of the expression of said additional DNA, exhibiting an increased resistance, as compared to a whole plant of the same plant species that is not so transformed, to tomato spotted wilt virus and another tospovirus, or propagation material of said transformed whole plant.

40. A transformed plant part, said plant part containing in its genome DNA in addition to the DNA naturally constituting the genome of said plant part, said plant part expressing said additional DNA, said additional DNA consisting of SEQ ID No:

1, and said transformed plant part, as a result of the expression of said additional DNA, exhibiting an increased resistance, as compared to a plant part of the same plant species and part type that is not so transformed, to tomato spotted wilt virus and another tospovirus, or propagation material of said transformed plant part.

42. A transformed plant part, said plant part containing in its genome DNA in addition to the DNA naturally constituting the genome of said plant part, said plant part expressing said additional DNA, said additional DNA consisting of SEQ ID No:

5, and said transformed plant part, as a result of the expression of said additional DNA, exhibiting an increased resistance, as compared to a plant part of the same plant species and part type that is not so transformed, to tomato spotted wilt virus and another tospovirus, or propagation material of said transformed plant part.