The Plant Pathology Journal

The Korean Society of Plant Pathology (한국식물병리학회)
권호 표지
DOI QR코드

DOI QR Code

The Application of RNA Transcript Conformation Polymorphism in Resolving Mixed Infection of PVY Isolates

  • Published : 2004.12.31
Download PDF
⟨ Previous Next ⟩

Abstract

A method based on RNA-transcript conformation polymorphism (TCP) was tested for detection of two PVY isolates in a mixed infection. Differences in electrophoretic mobility of RNA transcripts copied from PCR products of each virus isolate enabled the distinction between the two virus isolates in a mixed infection. The identities of the RNA transcripts and hence of the infecting virus isolates were determined by annealing to reference oligonucleotides containing unique strain-specific sequences visualized by retardation of transcript mobility in gel. The ratio at which both virus isolates could be detected was as low as 1:10. The suitability of this procedure for the study of mixed virus infections is discussed.

Keywords

References

  1. Barnett, O. W. 1992, A summary ofpotyvirus taxonomy and definitions. Arch. Virol. (Stipp.) 5:435-444 https://doi.org/10.1007/978-3-7091-6920-9_45
  2. De Bokx, J. A. and Huttinga, H. 1981. Potato virus Y. CMI/AAB, Description of Plant Viruses, No. 242
  3. Glais, L., Kerlan, C., Tribodet, M., Tordo, M-J. V., Robaglia, C. and Astier- Manifacier, S. 1996. Molecular characterization of potato virus $\Y^N$ isolates by PCR-RFLP. Eur. J Plant Pathol. 102:655-662 https://doi.org/10.1007/BF01877246
  4. Glais, L., Tribodet, M. and Kerlan, C. 2002. Genomic variability in potato potyvirus Y (PVY): evidence that PVY$\^N$W and PV$\Y^N^T^N$ variants are single to multiple recombinants betweenPVN$\^N$ isolates. Arch. Virol. 147:363-378 https://doi.org/10.1007/s705-002-8325-0
  5. Magom, H., Yoshikaw, N. and Takahashi, T. 1999. Singlestranded conformation polymorphism analysis of apple stem grooving capillovirus sequence variants. Phytopathology 89:136-140 https://doi.org/10.1094/PHYTO.1999.89.2.136
  6. Rosner, A. and Maslenin, L. 1999a. Differentiating $\PVY^{NTN}$ by unique single-restriction cleavage of PCR products. Potato Res. 42:215-221 https://doi.org/10.1007/BF02358411
  7. Rosner, A. and Maslenin, L. 1999b. Transcript conformation polymorphism: a novel approach for differentiating $\PVY^{NTN}$ . J Phytopathol. 147:661-664 https://doi.org/10.1046/j.1439-0434.1999.00055.x
  8. Rosner, A. and Maslenin, M. 2001. Improved electrophoretic resolution of RNA transcripts by specific homologous oligonucleotide gel retardation. J. Virol. Meth. 92: 1-4 https://doi.org/10.1016/S0166-0934(00)00218-4
  9. Rosner, A. and Maslenin, L. 2003. Tagging of viral RNA transcripts with strain-specific oligonucleotides: characterization and appIication. J. Virol. Meth. 110:105-109 https://doi.org/10.1016/S0166-0934(03)00107-1
  10. Rosner, A., Maslenin, L. and Spiegel, S. 1998. Differentiation among isolates of prunus necrotic ringspot virus by transcript conformation polymorphism. J Virol. Meth. 74:109-115 https://doi.org/10.1016/S0166-0934(98)00076-7
  11. Singh, N. and Singh, R. P. 1996. Nucleotide sequence and genomic organization of a Canadian isolate of the common strain of potato virus Y $\(PVY^o).$ Canad. J Plant Pathol. 18:209-224
  12. Spiegel, S., Scott, S. W., Bowman-Vance, V., Tam, Y., Galiakparov, N. N. and Rosner, A. 1996. Improved detection of prunus necrotic ringspot virus by the polymerase chain reaction. Eur. J. Plant Pathol. 102:681-685 https://doi.org/10.1007/BF01877249
  13. Spiegel, S., Tam, Y., Maslenin, L., Kolber, M., Nemeth, M. and Rosner, A.1999. Typing prunus necrotic virus by serology and restriction endonuclease analysis of PCR products. Ann. Appl. BioI. 135:395-400 https://doi.org/10.1111/j.1744-7348.1999.tb00866.x
  14. Tordo, M. J. V., Chachulska, A. M., Fakhfakh, H., Le Romancer, M., Robaglia, C. and Astier-Manifacier, S. 1995. Sequence polymorphism in the 5-UTR and in the P1 coding region of potato virus Y genomic RNA. J Gen. Virol. 70: 1951-1960 https://doi.org/10.1099/0022-1317-70-8-1951
  15. van den Heuvel, J. F. J. M., van der Vlugt, R. A. A., Verbeek, M., de Haan, O. T. and Huttinga, H. 1994. Characteristics ofresistance- breaking isolate of potato virus Ycausing potato tuber necrotic ringspot disease. Eur. J Plant Pathol' 100:347-356 https://doi.org/10.1007/BF01876445
  16. Weidemann, H. L. and Maiss, E. 1996. Detection of the potato tuber necrotic ringspotstrain of potato virus Y $\(PVY^{NTN})$ by reverse transcription and immunocapture polymerase chain reaction. J Plant Dis. Prot. 103:337-345