Skip to main content
SpringerLink
Log in

High variability in the N terminus of coat protein among potyviruses and its advantage in producing a specific antibody

  • Published:
European Journal of Plant Pathology Aims and scope Submit manuscript

Abstract

Cross reaction often occurs in serological detection for potyviruses that can lead to false positive results. Sequence alignment revealed that the N-terminal sequences of coat proteins (CP) are highly variable among potyviruses. Based on this finding, the preparation of a specific antibody against potyviruses is described here increasing the efficiency of detection and identification. A case study of Turnip mosaic virus (TuMV): a specific fragment “CP-50” (1–50 aa of CP) was obtained by prokaryotic expression and used for producing the polyclonal antibody. Test of specificity indicated that the prepared antibody “Anti-CP-50” strongly reacted with TuMV and is appropriate as a specific anti-TuMV antibody.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Abbreviations

TuMV:

Turnip mosaic virus

CP:

Coat protein

PAbs:

Polyclonal antibodies

MAbs:

Monoclonal antibodies

MW:

Molecular weight

ELISA:

Enzyme linked immunosorbent assay

GRAVY:

Grand average of hydropathicity.

References

  • Clark, M. F. (1981). Immunosorbent assays in plant pathology. Annual Review of Phytopathology, 19, 83–106.

    Article  CAS  Google Scholar 

  • Cooper, H. M., & Paterson, Y. (1995). Assays for antibody production. Current Protocols in Immunology, 2(4), 1–2.4.9.

    Google Scholar 

  • Gasteiger, E., Hoogland, C., Gattiker, A., Duvaud, S., Wilkins, M. R., Appel, R. D., et al. (2005). Protein identification and analysis tools on the ExPASy server. In J. M. Walker (Ed.), The proteomics protocols handbook. Totowa: Humana Press.

    Google Scholar 

  • Guruprasad, K., Reddy, B. V., & Pandit, M. W. (1990). Correlation between stability of a protein and its dipeptide composition: A novel approach for predicting in vivo stability of a protein from its primary sequence. Protein Engineering, 4, 155–161.

    Article  PubMed  CAS  Google Scholar 

  • Hong, J., Li, D. B., & Zhou, X. P. (2001). Classification atlas of plant virus. Beijing: Science Press.

    Google Scholar 

  • Hsu, H. (2009). Development of enzyme linked, tissue blot and dot blot immunoassays for plant virus detection. Plant Pathology, 508, 15–25.

    Article  CAS  Google Scholar 

  • ICTV. (2017). Virus Taxonomy: The Classification and Nomenclature of Viruses: The Online (10th) Report of the ICTV. http://talk.ictvonline.org/ictv-reports/ictv_online_report.

  • Kantrong, S., & Sako, N. (1993). Characterization of epitopes recognized by monoclonal antibodies to aphid transmissible and non-transmissible strains of Turnip mosaic virus. Archives of Virology, 133, 11–20.

    Article  PubMed  CAS  Google Scholar 

  • Kantrong, S., Saunal, H., Briand, J. P., & Sako, N. (1995). A single amino acid substitution at N-terminal region of protein of Turnip mosaic virus alters antigenicity and aphid transmissibility. Virology, 140, 453–467.

    CAS  Google Scholar 

  • Kyte, J., & Doolittle, R. F. (1982). A simple method for displaying the hydropathic character of a protein. Journal of Molecular Biology, 157, 105–132.

    Article  PubMed  CAS  Google Scholar 

  • Naidu, R. A., & Hughes, J. d’A. (1997). Methods for the detection of plant virus diseases. International Institute of Tropical Agriculture, Ibadan, Nigeria, 45: 1–28.

  • Nguyen, H. D., Tomitaka, Y., Ho, S. Y., Duchêne, S., Vetten, H. J., Lesemann, D., et al. (2013). Turnip mosaic potyvirus probably first spread to eurasian brassica crops from wild orchids about 1000 years ago. PLoS One, 8(2), e55336.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Pearson, W. R., & Lipman, D. J. (1988). Improved tools for biological sequence comparison. Proceedings of the National Academy of Sciences, 85, 2444–2448.

    Article  CAS  Google Scholar 

  • Shukla, D. D., & Ward, C. W. (1989). Structure of potyvirus coat proteins and its application in the taxonomy of the potyvirus group. Advances in Virus Research, 36, 273–314.

    Article  PubMed  CAS  Google Scholar 

  • Shukla, D. D., Strike, P. M., Tracy, S. L., Gough, K. H., & Ward, C. W. (1988). The N and C termini of the coat proteins of potyviruses are surface-located and the N terminus contains the major virus-specific epitopes. Journal of General Virology, 69, 1497–1508.

    Article  CAS  Google Scholar 

  • Shukla, D. D., Jilka, J., Tosic, M., & Ford, R. E. (1989a). A novel approach to the serology of potyviruses involving affinity purified polyclonal antibodies directed towards virus-specific N termini of coat proteins. Journal of General Virology, 70, 13–23.

    Article  CAS  Google Scholar 

  • Shukla, D. D., Tribbick, G., Mason, T. J., Hewish, D. R., Geysen, H. M., & Ward, C. W. (1989b). Localization of virus-specific and group-specific epitopes of plant potyviruses by systematic immunochemical analysis of overlapping peptide fragments. Proceedings of the National Academy of Sciences of the United States of America, 86, 8192–8196.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Thompson, S., Fraser, R. S., & Barnden, K. L. (1988). A beneficial effect of trypsin on the purification of Turnip mosaic virus (TuMV) and other potyviruses. Journal of Virological Methods, 20, 57–64.

    Article  PubMed  CAS  Google Scholar 

  • Walsh, J. A., & Jenner, C. E. (2002). Turnip mosaic virus and the quest for durable resistance. Molecular Plant Pathology, 3, 289–300.

    Article  PubMed  CAS  Google Scholar 

  • Zheng, G. H., Peng, D. W., Tong, Q. X., Zheng, Z. Z., & Ming, Y. L. (2017). Occurrence of Turnip mosaic virus in Phalaenopsis sp. in China. Journal of Plant Pathology, 99, 703–706.

    Google Scholar 

Download references

Acknowledgements

The authors thank Prof. Chang Chin-an (Graduate Institute of Biochemical Sciences and Technology, Chaoyang University of Technology, Taichung, 41349, Taiwan) for providing experimental materials and technical assistance. We thank Peter Sol Reinach (Wenzhou Medical University) for providing language editing service of this manuscript. This work is supported by the grant from Fujian Provincial Natural Science Foundation of China (2011D002) and Xiamen Municipal Natural Science Foundation (3502Z20127015 & 3502Z20162014).

Funding

Fujian Provincial Natural Science Foundation of China (2011D002); Xiamen Municipal Natural Science Foundation (3502Z20127015 & 3502Z20162014).

Author information

Author notes

  1. D. W. Peng and G. H. Zheng contributed equally to the work.

Authors and Affiliations

  1. Xiamen Overseas Chinese Subtropical Plant Introduction Garden, National Plant Introduction Quarantine Base, Xiamen Key Laboratory for Plant Introduction, Quarantine and Natural Products, Xiamen, 361002, Fujian, China

    Dewei Peng, Guohua Zheng, Zhizhong Zheng, Qingxuan Tong & Yanlin Ming

  2. School of Ophthalmology and Optometry, Eye Hospital, Wenzhou Medical University, Wenzhou, 325027, Zhejiang, China

    Dewei Peng

  3. College of Chemical Engineering, Huaqiao University, Xiamen, 361021, Fujian, China

    Qingxuan Tong & Yanlin Ming

Authors
  1. Dewei Peng
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Guohua Zheng
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Zhizhong Zheng
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Qingxuan Tong
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yanlin Ming
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

YLM and QXT conceived the experiments, and critically revised the manuscript. GHZ collected the samples. DWP and GHZ performed the experiments. ZZZ analyzed the data. DWP wrote the manuscript.

Corresponding author

Correspondence to Yanlin Ming.

Ethics declarations

This study was carried out following the guidelines and approval of the Laboratory Animal Ethics Committee of Wenzhou Medical University. The rabbits were anesthetized by 3% pentobarbital sodium (1 ml/kg) when collecting blood samples. After the experiments, rabbits were submitted to euthanasia using anesthetic overdose.

Conflict of interest

The authors declare that they have no competing interests.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Peng, D., Zheng, G., Zheng, Z. et al. High variability in the N terminus of coat protein among potyviruses and its advantage in producing a specific antibody. Eur J Plant Pathol 152, 385–393 (2018). https://doi.org/10.1007/s10658-018-1482-z

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10658-018-1482-z

Keywords

Search

Navigation