Skip to main content
SpringerLink
Log in

Monoclonal Antibody-Based Serological Detection Methods for Wheat Dwarf Virus

  • Research Article
  • Published:
Virologica Sinica

Abstract

Wheat dwarf disease caused by wheat dwarf virus (WDV) is currently present in wheat growing regions in China and causes serious losses in wheat yield. To develop reliable and effective serological detection methods for WDV, the coat protein (CP) gene of WDV was cloned and expressed in Escherichia coli. The purified recombinant CP protein was immunized to BALB/c mice, and four hybridoma cell lines (i.e. 18G10, 9G4, 23F4 and 22A10) secreting anti-WDV monoclonal antibodies (MAbs) were obtained through the hybridoma technique. Using the prepared MAbs, an antigen-coated-plate enzyme-linked immunosorbent assay (ACP-ELISA) and a dot-ELISA were established for detecting WDV in wheat samples. The most sensitive ACP-ELISA based on MAb 23F4 or 22A10 was able to detect WDV in 1:163,840 (w/v, g/mL) diluted WDV-infected wheat plant crude extracts. The dot-ELISA based on MAb 23F4 was the most sensitive and able to detect the virus in 1:5,120 (w/v, g/mL) diluted wheat plant crude extracts. A total of 128 wheat samples were collected from wheat growing regions in the Shaanxi and Qinghai provinces, China, and were screened for the presence of WDV using two developed serological assays. Results from the survey showed that approximately 62% of the samples were infected with WDV. PCR followed by DNA sequencing and sequence alignment validated the results from the two serological assays. Therefore, we consider that these two serological detection methods can be significantly useful for the control of WDV in China.

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.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Achon MA, Serrano L, Ratti C, Rubies-Autonell C (2006) First detection of wheat dwarf virus in barley in Spain associated with an outbreak of barley yellow dwarf. Plant Dis 90:970

    Article  Google Scholar 

  • Bendahmane M (1995) Identification and characterization of wheat dwarf virus from france using a rapid method for geminivirus DNA preparation. Phytopathology 85:1449–1455

    Article  CAS  Google Scholar 

  • Chen Z, Zhang MH, Zhou XP, Wu JX (2017) Development and detection application of monoclonal antibodies against Zucchini yellow mosaic virus. J Integr Agr 16:115–124

    Article  CAS  Google Scholar 

  • Ekzayez AM, Kumari SG, Ismail I (2010) First report of wheat dwarf virus and its vector (Psammotettix provincialis) affecting wheat and barley crops in Syria. Plant Dis 95:76

    Article  Google Scholar 

  • Gadiou S, Ripl J, Janourova B, Jarosova J, Kundu JK (2012) Real-time PCR assay for the discrimination and quantification of wheat and barley strains of wheat dwarf virus. Virus Genes 44:349–355

    Article  CAS  PubMed  Google Scholar 

  • Gawel NJ, Jarret RL (1991) A modified CTAB DNA extraction procedure for Musa and Ipomoea. Plant Mol Biol Rep 9:262–266

    Article  CAS  Google Scholar 

  • Jin W, Zhang J, Liu Y, Wang X (2015) Development and application of nucleic acid spot hybridization (NASH) assay for rapid detection of Wheat dwarf virus. Plant Protect 41:100–103 (in Chinese)

    CAS  Google Scholar 

  • Kapooria RG, Ndunguru J (2004) Occurrence of viruses in irrigated wheat in Zambia. EPPO Bull 34:413–419

    Article  Google Scholar 

  • Koklu G, Ramsell JN, Kvarnheden A (2007) The complete genome sequence for a Turkish isolate of wheat dwarf virus (WDV) from barley confirms the presence of two distinct WDV strains. Virus Genes 34:359–366

    Article  PubMed  Google Scholar 

  • Lemmetty A, Huusela-Veistola E (2005) First report of wheat dwarf virus in winter wheat in Finland. Plant Dis 89:912–912

    Article  Google Scholar 

  • Li N, Chen Z, Liu Y, Liu Y, Zhou XP, Wu JX (2015) Development of monoclonal antibodies and serological assays specific for Barley yellow dwarf virus GAV strain. Virol J 12:136

    Article  PubMed  PubMed Central  Google Scholar 

  • Lindblad M, Sigvald R (2004) Temporal spread of wheat dwarf virus and mature plant resistance in winter wheat. Crop Protect 23:229–234

    Article  Google Scholar 

  • Liu H, Song XJ, Ni YQ, Lu LN, Zhou XP, Wu JX (2014) Highly sensitive and specific monoclonal antibody-based serological methods for rice ragged stunt virus detection in rice plants and rice brown planthopper vectors. J Integr Agr 13:1943–1951

    Article  CAS  Google Scholar 

  • Liu Z, Chen Z, Hong J, Wang XF, Zhou CY, Zhou XP, Wu JX (2016) Monoclonal antibody-based serological methods for detecting Citrus tristeza virus in citrus groves. Virol Sin 31:324–330

    Article  CAS  PubMed  Google Scholar 

  • Liu Z, SunZhu YJ, Zhou XP, Hong J, Wu JX (2017) Monoclonal antibody-based serological detection of Citrus yellow vein clearing virus in citrus groves. J Integr Agr 16:884–891

    Article  CAS  Google Scholar 

  • Nygren J, Shad N, Kvarnheden A, Westerbergh A (2015) Variation in susceptibility to wheat dwarf virus among wild and domesticated wheat. PLoS ONE 10:e0121580

    Article  PubMed  PubMed Central  Google Scholar 

  • Ramsell JNE, Lemmetty A, Jonasson J, Andersson A, Sigvald R, Kvarnheden A (2008) Sequence analyses of Wheat dwarf virus isolates from different hosts reveal low genetic diversity within the wheat strain. Plant Pathol 57:834–841

    Article  CAS  Google Scholar 

  • Schalk HJ, Matzeit V, Schiller B, Schell J, Gronenborn B (1989) Wheat dwarf virus, a geminivirus of graminaceous plants needs splicing for replication. EMBO J 8:359–364

    CAS  PubMed  PubMed Central  Google Scholar 

  • Shang HL, Xie Y, Zhou XP, Qian YJ, Wu JX (2011) Monoclonal antibody-based serological methods for detection of Cucumber green mottle mosaic virus. Virol J 8:228

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Vacke J (1961) Wheat dwarf virus disease. Biol Plantarum 3:228–233

    Google Scholar 

  • Wang Y, Mao Q, Liu W, Mar T, Wei T, Liu Y, Wang X (2014) Localization and distribution of Wheat dwarf virus in its vector leafhopper, Psammotettix alienus. Phytopathol 104:897–904

    Article  CAS  Google Scholar 

  • Wang L, Liu Y, Wang XF (2016) Detection of Wheat dwarf virus by TaqMan LNA probe real-time PCR. Acta Phytopathologica Sin 46:313–319 (in Chinese)

    Google Scholar 

  • Wu J, Ni Y, Liu H, Rao L, Zhou Y, Zhou X (2013) Development and use of three monoclonal antibodies for the detection of rice black-streaked dwarf virus in field plants and planthopper vectors. Virol J 10:114

    Article  PubMed  PubMed Central  Google Scholar 

  • Wu J, Ni Y, Liu H, Ding M, Zhou X (2014) Monoclonal antibody-based serological assays and immunocapture-RT-PCR for detecting Rice dwarf virus in field rice plants and leafhopper vectors. J Virol Methods 195:134–140

    Article  CAS  PubMed  Google Scholar 

  • Wu B, Shang X, Schubert J, Habekuß A, Elena SF, Wang X (2015) Global-scale computational analysis of genomic sequences reveals the recombination pattern and coevolution dynamics of cereal-infecting geminiviruses. Sci Rep 5:8153

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Xie J, Wang X, Liu Y, Peng Y, Zhou G (2007) First report of the occurrence of wheat dwarf virus in wheat in China. Plant Dis 91:111

    Article  Google Scholar 

  • Zhang X, Zhou G, Wang X (2010) Detection of wheat dwarf virus (WDV) in wheat and vector leafhopper (Psammotettix alienus Dahlb.) by real-time PCR. J Virol Methods 169:416–419

    Article  CAS  PubMed  Google Scholar 

  • Zhou X, Xie Y, Peng Y, Zhang Z (2003) Malvastrum yellow vein virus, a newBegomovirus species associated with satellite DNA molecule. Chin Sci Bull 48:2206–2210

    Article  Google Scholar 

Download references

Acknowledgements

We are grateful to Dr. Xinshun Ding (Samuel Roberts Noble Foundation, Ardmore, USA) for his valuable comments and manuscript edits. This work was supported by Public Science and Technology Research Funds Projects of Agriculture (201303021), and the National Basic Research Program (973) of China (No. 2014CB138400).

Author information

Authors and Affiliations

  1. State Key Laboratory of Rice Biology, Institute of Biotechnology, Zhejiang University, Hangzhou, 310058, China

    Minghao Zhang, Rui Chen, Xueping Zhou & Jianxiang Wu

  2. State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, 100193, China

    Xueping Zhou

Authors
  1. Minghao Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Rui Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Xueping Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jianxiang Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Author Contributions

MHZ prepared the MAbs and carried out the immunoassays. RC performed the PCR detection of wheat samples. XPZ and JXW conceived of the study, participated in its design and helped to draft the manuscript. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Xueping Zhou or Jianxiang Wu.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Animal and Human Rights Statement

The animal experiments were performed in accordance with the Principles of the Helsinki accords and approved by the Animal Experimentation Ethics Committee of Zhejiang University, Hangzhou, China.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Zhang, M., Chen, R., Zhou, X. et al. Monoclonal Antibody-Based Serological Detection Methods for Wheat Dwarf Virus. Virol. Sin. 33, 173–180 (2018). https://doi.org/10.1007/s12250-018-0024-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12250-018-0024-3

Keywords

Search

Navigation