Detection of Rupestris stem pitting associated virus

• US 6,395,490 B1
• Filed: 05/18/2000
• Issued: 05/28/2002
• Est. Priority Date: 05/20/1997
• Status: Expired due to Fees

• Current Assignee: Cornell Research Foundation Incorporated (Cornell University)
• Original Assignee: Cornell Research Foundation Incorporated (Cornell University)
• Inventors: Gonsalves, Dennis, Meng, Baozhong
• Primary Examiner(s): Jones, W. Gary
• Assistant Examiner(s): Tung, Joyce
• Application Number:
  US09/574,141
• Time in Patent Office: 740 Days
• Field of Search: 435/6,91.2 536/24.3
• US Class Current: 435/6.12
• CPC Class Codes: C07K 14/005 from viruses
• CPC Class Codes: C12N 15/8283 for virus resistance
• CPC Class Codes: C12N 2770/26022 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: C12Q 1/701 Specific hybridization probes
• CPC Class Codes: Y10S 424/801 involving antibody or fragment thereof produced by recombinant dna technology

Abstract

The present invention relates to an isolated protein or polypeptide corresponding to a protein or polypeptide of a Rupestris stem pitting associated virus. The encoding DNA molecule, either alone in isolated form, in an expression system, a host cell, or a transgenic grape plant, is also disclosed. Other aspects of the present invention relate to a method of imparting Rupestris stem pitting associated virus resistance to grape plants by transforming them with the DNA molecule of the present invention, and a method of detecting the presence of a Rupestris stem pitting associated virus, such as RSPaV-1, in a sample.

Claims

1. A method for determining whether a sample includes a Rupestris stem pitting associated virus, said method comprising:

contacting the sample with a Rupestris stem pitting associated virus-specific nucleic acid probe under conditions that allow for hybridization of the probe to a Rupestris stem pitting associated virus nucleic acid molecule, wherein a detectable hybridization signal indicates that Rupestris stem pitting associated virus is present in the sample.

2. The method of claim 1, wherein hybridization is detected using a dot blot hybridization assay, tissue printing assay, southern hybridization assay, northern hybridization assay, or gene amplification detection procedure.

3. The method of claim 1, wherein said nucleic acid probe comprises a nucleic acid molecule that hybridizes with a nucleic acid molecule encoding an open reading frame of a Rupestris stem pitting associated virus protein or polypeptide.

4. The method of claim 1, wherein said sample comprises plant material.

5. The method of claim 4, wherein said plant material comprises a plant cell or tissue.

6. The method of claim 4, wherein said plant material comprises grape plant material.

7. The method of claim 6, wherein said grape plant material comprises a rootstock or scion.

8. The method of claim 4, wherein said plant material comprises propagated plant material.

9. The method of claim 1, wherein said method detects RSPaV-1, RSP47-4, or RSP158.

10. The method of claim 1, wherein said nucleic acid probe comprises SEQ. ID. No. 51, SEQ. ID. No. 52, SEQ. ID. No. 53, or SEQ. ID. No. 54.

11. A method for determining whether a Rupestris stem pitting associated virus is present in a sample, said method comprising:

contacting the sample with a Rupestris stem pitting associated virus-specific nucleic acid probe under conditions that allow for hybridization of the probe to a Rupestris stem pitting associated virus nucleic acid molecule, wherein an undetectable hybridization signal indicates that Rupestris stem pitting associated virus is not present in the sample.

12. The method of claim 11, wherein hybridization is detected using a dot blot hybridization assay, tissue printing assay, southern hybridization assay, northern hybridization assay, or gene amplification detection procedure.

13. The method of claim 11, wherein said nucleic acid probe comprises a nucleic acid molecule that hybridizes with a nucleic acid molecule encoding an open reading frame of a Rupestris stem pitting associated virus protein or polypeptide.

14. The method of claim 11, wherein said sample comprises plant material.

15. The method of claim 14, wherein said plant material comprises a plant cell or tissue.

16. The method of claim 14, wherein said plant material comprises grape plant material.

17. The method of claim 16, wherein said grape plant material comprises a rootstock or scion.

18. The method of claim 14, wherein said plant material comprises propagated plant material.

19. The method of claim 11, wherein said nucleic acid probe comprises SEQ. ID. No. 51, SEQ. ID. No. 52, SEQ. ID. No. 53, or SEQ. ID. No. 54.

20. A method of determining whether a plant is infected with a Rupestris stem pitting associated virus, said method comprising the steps of:

(a) providing a sample from said plant; and

(b) contacting the sample with a nucleic acid probe comprising a Rupestris stem pitting virus nucleic acid molecule under conditions that allow for the hybridization of the probe to a Rupestris stem pitting associated virus nucleic acid molecule, wherein a detectable hybridization signal indicates that the plant is infected with Rupestris stem pitting associated virus.

21. The method of claim 20, further comprising selecting a plant that is infected with a Rupestris stem pitting associated virus.

22. The method of claim 20, wherein said hybridization is detected by using a dot blot hybridization assay, tissue printing assay, southern hybridization assay, northern hybridization assay, or gene amplification detection procedure.

23. The method of claim 20, wherein said nucleic acid probe comprises a nucleic acid molecule that hybridizes with a nucleic acid molecule encoding an open reading frame of a Rupestris stem pitting associated virus protein or polypeptide.

24. The method of claim 20, wherein said sample comprises plant material.

25. The method of claim 24, wherein said plant material comprises a plant cell or tissue.

26. The method of claim 24, wherein said plant material comprises grape plant material.

27. The method of claim 26, wherein said grape plant material comprises a rootstock or scion.

28. The method of claim 24, wherein said plant material comprises propagated plant material.

29. The method of claim 20, wherein said method detects RSPaV-1, RSP47-4,or RSP158.

30. The method of claim 20, wherein said nucleic acid probe comprises SEQ. ID. No. 51, SEQ. ID. No. 52, SEQ. ID. No. 53, or SEQ. ID. No. 54.

31. A method of determining whether a plant is infected with a Rupestris stem pitting associated virus, said method comprising the steps of:

(a) providing a sample from said plant; and

(b) contacting the sample with a nucleic acid probe comprising a Rupestris stem pitting virus nucleic acid molecule under conditions that allow for the hybridization of the probe to a Rupestris stem pitting associated virus nucleic acid molecule, wherein an undetectable hybridization signal indicates that the plant is not infected with a Rupestris stem pitting associated virus.

32. The method of claim 31, further comprising selecting the plant that is not infected with a Rupestris stem pitting associated virus.

33. The method of claim 31, wherein said hybridization is detected by using a dot blot hybridization assay, tissue printing assay, southern hybridization assay, northern hybridization assay, or gene amplification detection procedure.

34. The method of claim 31, wherein said nucleic acid probe comprises a nucleic acid molecule that hybridizes with a nucleic acid molecule encoding an open reading frame of a Rupestris stem pitting associated virus protein or polypeptide.

35. The method of claim 31, wherein said sample comprises plant material.

36. The method of claim 35, wherein plant material comprises a plant cell or tissue.

37. The method of claim 35, wherein said plant material comprises grape plant material.

38. The method of claim 37, wherein said grape plant material comprises a rootstock or scion.

39. The method of claim 35, wherein said plant material comprises propagated plant material.

40. The method of claim 31, wherein said nucleic acid probe comprises SEQ. ID. No. 51, SEQ. ID. No. 52, SEQ. ID. No. 53, or SEQ. ID. No. 54.