Development of Soybean Yellow Mottle Mosaic Virus-Based Expression Vector for Heterologous Protein Expression in French Bean
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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.
KeywordsFrench bean GFP expression Plant viral vector Heterologous gene expression Soybean yellow mottle mosaic virus
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.
- 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
- 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.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
- 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
- 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