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Development of Soybean Yellow Mottle Mosaic Virus-Based Expression Vector for Heterologous Protein Expression in French Bean

  • Nagamani SandraEmail author
  • A Abdul Kader Jailani
  • Rakesh Kumar Jain
  • Bikash Mandal
Original Paper
  • 55 Downloads

Abstract

Plant virus-based vectors provide attractive and valuable tools for rapid production of recombinant protein in large quantities as they produce systemic infections in differentiated plant tissues. In the present study, we engineered the Soybean yellow mottle mosaic virus (SYMMV) as a gene expression vector which is a promising candidate for systemic expression of foreign proteins in French bean plants. Full virus vector strategy was exploited for insertion of foreign gene by inserting MCS through PCR in the circular pJET-SYMMV clone. To examine the ability of the SYMMV vector system, GFP gene was cloned after the start codon of coat protein (CP) so that its expression was driven by the SYMMV-CP subgenomic promoter. When in vitro run off SYMMV-GFP transcript was mechanically inoculated to French bean leaves, good level of GFP expression was observed through confocal microscopy up to 40 dpi. Expression of heterologous protein was also confirmed through ISEM, DAC-ELISA and RT-PCR with specific primers at 20 dpi. The recombinant SYMMV construct was stable in in vitro runoff transcript inoculated plants but the inserted GFP was lost in progeny virion inoculated plants. The system developed here will be useful for further studies of SYMMV gene functions and exploitation of SYMMV as a gene expression vector.

Keywords

French bean GFP expression Plant viral vector Heterologous gene expression Soybean yellow mottle mosaic virus 

Notes

Acknowledgements

The INSPIRE fellowship to the first author provided by Department of Science and Technology (DST), New Delhi, India, financial support by National Agricultural Science Fund (NASF), ICAR and controlled environmental conditions provided by National Phytotron Facility (NPF) are thankfully acknowledged.

References

  1. 1.
    Baek, I. Y., Choi, H. S., Kim, J. S., Kim, K. H., Lee, S. H., Moon, J. S., et al. (2012). Recombinant virus-induced gene silencing vector from SYMMV useful for functional analysis of useful genes in soybean and uses thereof. Publication No. WO2012053710 A1; Application No. PCT/KR2011/003161.Google Scholar
  2. 2.
    Baulcombe, D. C., Chapman, S., & Santa Cruz, S. (1995). Jellyfish green fluorescent protein as a reporter for plant virus infections. Plant Journal, 7, 1045–1053.CrossRefGoogle Scholar
  3. 3.
    Burch-Smith, T. M., Anderson, J. C., Martin, G. B., & Dinesh-Kumar, S. P. (2004). Applications and advantages of virus-induced gene silencing for gene function studies in plants. Plant Journal, 39(5), 734–746.CrossRefGoogle Scholar
  4. 4.
    Constantin, G. D., Krath, B. N., MacFarlane, S. A., Nicolaisen, M., Johansen, I. E., & Lund, O. S. (2004). Virus-induced gene silencing as a tool for functional genomics in a legume species. Plant Journal, 40, 622–631.CrossRefGoogle Scholar
  5. 5.
    Diaz-Camino, C., Annamalai, P., Sanchez, F., Kachroo, A., & Ghabrial, S. A. (2011). An effective virus-based gene silencing method for functional genomics studies in common bean. Plant Methods, 7, 16.CrossRefGoogle Scholar
  6. 6.
    Gleba, Y., Marillonnet, S., & Klimyuk, V. (2004). Engineering viral expression vectors for plants: The ‘full virus’ and the ‘deconstructed virus’ strategies. Current Opinion in Plant Biology, 7, 182–188.CrossRefGoogle Scholar
  7. 7.
    Gleba, Y., Klimyuk, V., & Marillonnet, S. (2007). Viral vectors for the expression of proteins in plants. Current Opinion in Biotechnology, 18, 134–141.CrossRefGoogle Scholar
  8. 8.
    Gleba, Y. Y., Tuse, D., & Giritch, A. (2014). Plant viral vectors for delivery by Agrobacterium. Current Topics in Microbiology and Immunology, 375, 155–192.PubMedGoogle Scholar
  9. 9.
    Hacker, D. L., Petty, I. T. D., Wei, N., & Morris, T. J. (1992). Turnip crinkle virus genes required for RNA replication and virus movement. Virology, 186, 1–8.CrossRefGoogle Scholar
  10. 10.
    Heaton, L. A., Lee, T. C., Wei, N., & Morris, T. J. (1991). Point mutations in the turnip crinkle virus capsid protein affect symptoms expressed by Nicotiana benthamiana. Virology, 183, 143–150.CrossRefGoogle Scholar
  11. 11.
    Hefferon, K. (2014). Plant virus expression vector development: New perspectives. BioMed Research International.  https://doi.org/10.1155/2014/785382.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Ido, Y., Nakahara, K. S., & Uyeda, I. (2012). White clover mosaic virus-induced gene silencing in pea. Journal of General Plant Pathology, 78, 127–132.CrossRefGoogle Scholar
  13. 13.
    Igarashi, A., Yamagata, K., Sugai, T., Takahashi, Y., Sugawara, E., Tamura, A., et al. (2009). Apple latent spherical virus vectors for reliable and effective virus-induced gene silencing among a broad range of plants including tobacco, tomato, Arabidopsis thaliana, cucurbits and legumes. Virology, 386, 407–416.CrossRefGoogle Scholar
  14. 14.
    Koenig, R., Lesemann, D. E., Loss, S., Engelmann, J., Commandeur, U., Dem, G., et al. (2006). Zygocactus virus X-based expression vectors and formation of rod-shaped virus-like particles in plants by the expressed coat proteins of Beet necrotic yellow vein virus and Soil-borne cereal mosaic virus. Journal of General Virology, 87, 439–443.CrossRefGoogle Scholar
  15. 15.
    Lefkowitz, E. J., Dempsey, D. M., Hendrickson, R. C., Orton, R. J., Siddell, S. G., & Smith, D. B. (2018). Virus taxonomy: The database of the International Committee on Taxonomy of Viruses (ICTV). Nucleic Acids Research, 46, D708–D717.CrossRefGoogle Scholar
  16. 16.
    Li, S., Moon, J. S., Lee, S. H., & Domier, L. L. (2009). First report of Soybean yellow mottle mosaic virus in soybean in North America. Plant Disease, 93, 1214.CrossRefGoogle Scholar
  17. 17.
    Lim, S., Nam, M., Kim, K. H., Lee, S. H., Moon, J. K., Lim, H. S., et al. (2015). Development of a new vector using Soybean yellow common mosaic virus for gene function study or heterologous protein expression in soybeans. Journal of Virological Methods, 228, 1–9.CrossRefGoogle Scholar
  18. 18.
    Lindbo, J. A. (2007). High-efficiency protein expression in plants from agroinfection-compatible Tobacco mosaic virus expression vectors. BMC Biotechnology, 7, 52.CrossRefGoogle Scholar
  19. 19.
    Liu, L., & Lomonossoff, G. P. (2002). Agroinfection as a rapid method for propagating Cowpea mosaic virus-based constructs. Journal of Virological Methods, 105, 343–348.CrossRefGoogle Scholar
  20. 20.
    Lommel, S. A., McCain, A. H., & Morris, T. J. (1982). Evaluation of indirect enzyme-linked immunosorbent assay for the detection of plant viruses. Phytopathology, 72, 1018–1022.CrossRefGoogle Scholar
  21. 21.
    Meng, Y., Moscou, M. J., & Wise, R. P. (2009). Blufensin1 negatively impacts basal defense in response to barley powdery mildew. Plant Physiology, 149, 271–285.CrossRefGoogle Scholar
  22. 22.
    Nagamatsu, A., Masuta, C., Senda, M., Matsuura, H., Kasai, A., Hong, J. S., et al. (2007). Functional analysis of soybean genes involved in flavonoid biosynthesis by virus-induced gene silencing. Plant Biotechnology Journal, 5, 778–790.CrossRefGoogle Scholar
  23. 23.
    Nam, M., Kim, S. M., Domier, L. L., Koh, S., Moon, J. K., Choi, H. S., et al. (2009). Nucleotide sequence and genome organization of a newly identified member of the genus Carmovirus, Soybean yellow mottle mosaic virus from soybean. Archives of Virology, 154, 1679–1684.CrossRefGoogle Scholar
  24. 24.
    Pflieger, S., Blanchet, S., Meziadi, C., Richard, M. M. S., Thareau, V., Mary, F., et al. (2014). The “one-step” Bean pod mottle virus (BPMV)derived vector is a functional genomics tool for efficient over expression of heterologous protein, virus-induced gene silencing and genetic mapping of BPMV R-gene in common bean (Phaseolus vulgaris L.). BMC Plant Biology, 14, 1–16.CrossRefGoogle Scholar
  25. 25.
    Pogue, G. P., Lindbo, J. A., Garger, S. J., & Fitzmaurice, W. P. (2002). Making an ally from an enemy: Plant virology and the new agriculture. Annual Reviews of Phytopathology, 40, 45–74.CrossRefGoogle Scholar
  26. 26.
    Sandra, N., Jailani, A. A. K., Jain, R. K., & Mandal, B. (2017). Genome characterization, infectivity assays of in vitro and in vivo infectious transcripts of soybean yellow mottle mosaic virus from India reveals a novel short mild genotype. Virus Research, 232, 96–105.CrossRefGoogle Scholar
  27. 27.
    Sandra, N., Kumar, A., Sharma, P., Kapoor, R., Jain, R. K., & Mandal, B. (2015). Diagnosis of a new variant of soybean yellow mottle mosaic virus with extended host-range in India. Virus Disease, 26(4), 304–314.CrossRefGoogle Scholar
  28. 28.
    Sandra, N., Tripathi, A., Lal, S. K., Kumar, A., Mandal, B., & Jain, R. K. (2018). First report of Soybean yellow mottle mosaic virus on Soybean (Glycine max) in India. Plant Disease, 102(8), 1673.Google Scholar
  29. 29.
    Scholthof, H. B., Scholthof, K. B., & Jackson, A. O. (1996). Plant virus gene vectors for transient expression of foreign proteins in plants. Annual Review of Phytopathology, 34, 299–323.CrossRefGoogle Scholar
  30. 30.
    Seo, J. K., Lee, H. G., & Kim, K. H. (2009). Systemic gene delivery into soybean by simple rub-inoculation with plasmid DNA of a Soybean mosaic virus-based vector. Archives of Virology, 154, 87–99.CrossRefGoogle Scholar
  31. 31.
    Shivprasad, S., Pogue, G. P., Lewandowski, D. J., Hidalgo, J., Donson, J., Grill, L. K., et al. (1999). Heterologous sequences greatly affect foreign gene expression in tobacco mosaic virus-based vectors. Virology, 255, 312–323.CrossRefGoogle Scholar
  32. 32.
    Toth, R. L., Pogue, G. P., & Chapman, S. (2002). Improvement of the movement and host range properties of a plant virus vector through DNA shuffling. Plant Journal, 30, 593–600.CrossRefGoogle Scholar
  33. 33.
    Tuttle, J. R., Haigler, C. H., & Robertson, D. (2012). Method: Low-cost delivery of the cotton leaf crumple virus-induced gene silencing system. Plant Methods, 8, 27.CrossRefGoogle Scholar
  34. 34.
    Varallyay, E., Lichner, Z., Safrany, J., Havelda, Z., Salamon, P., Bisztray, G., et al. (2010). Development of a virus induced gene silencing vector from a legumes infecting tobamovirus. Acta Biologica Hungarica, 61, 457–469.CrossRefGoogle Scholar
  35. 35.
    Yang, C. D., Liao, J. T., Lai, C. Y., Jong, M. H., Liang, C. M., Lin, Y. L., et al. (2007). Induction of protective immunity in swine by recombinant bamboo mosaic virus expressing foot-and-mouth disease virus epitopes. BMC Biotechnology, 7, 62.CrossRefGoogle Scholar
  36. 36.
    Youssef, F., Marais, A., Faure, C., Gentit, P., & Candresse, T. (2011). Strategies to facilitate the development of uncloned or cloned infectious full-length viral cDNAs: Apple chlorotic leaf spot virus as a case study. Virology Journal, 8, 488–500.CrossRefGoogle Scholar
  37. 37.
    Zhang, C., & Ghabrial, S. A. (2006). Development of Bean pod mottle virus-based vectors for stable protein expression and sequence-specific virus-induced gene silencing in soybean. Virology, 344, 401–411.CrossRefGoogle Scholar
  38. 38.
    Zhang, C., Bradshaw, J. D., Whitham, S. A., & Hill, J. H. (2010). The development of an efficient multipurpose Bean pod mottle virus viral vector set for foreign gene expression and RNA silencing. Plant Physiology, 153(1), 52–65.CrossRefGoogle Scholar
  39. 39.
    Zhang, C., Whitham, S. A., & Hill, J. H. (2013). Virus-induced gene silencing in soybean and common bean. In A. Becker (Ed.), Virus-induced gene silencing: Methods and protocols, methods in molecular biology (pp. 149–156). New York: Springer.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Division of Seed Science and TechnologyIndian Agricultural Research InstituteNew DelhiIndia
  2. 2.Advanced Centre for Plant Virology, Division of Plant PathologyIndian Agricultural Research InstituteNew DelhiIndia

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