Adoption of the 2A Ribosomal Skip Principle to Track Assembled Virions of Pepper Mild Mottle Virus in Nicotiana benthamiana

Mengting Jiao; Yueyan Yin; Yanzhen Tian; Jianing Lei; Lin Lin; Jian Wu; Yuwen Lu; Hongying Zheng; Fei Yan; Jianguang Wang; Jiejun Peng

doi:10.3390/plants13070928

Adoption of the 2A Ribosomal Skip Principle to Track Assembled Virions of Pepper Mild Mottle Virus in Nicotiana benthamiana

Plants (Basel). 2024 Mar 22;13(7):928. doi: 10.3390/plants13070928.

Authors

Mengting Jiao^{1

2

3}, Yueyan Yin⁴, Yanzhen Tian^{1

2}, Jianing Lei^{1

2}, Lin Lin^{1

2}, Jian Wu^{1

2}, Yuwen Lu^{1

2}, Hongying Zheng^{1

2}, Fei Yan^{1

2}, Jianguang Wang³, Jiejun Peng^{1

2}

Affiliations

¹ State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agroproducts, Institute of Plant Virology, Ningbo University, Ningbo 315211, China.
² Key Laboratory of Biotechnology in Plant Protection of MARA and Zhejiang Province, Institute of Plant Virology, Ningbo University, Ningbo 315211, China.
³ Biocontrol Engineering Research Center of Crop Disease & Pest of Yunnan Province, School of Ecology and Environmental Science, Yunnan University, Kunming 650091, China.
⁴ Institute of Biotechnology and Germplasm Resources, Yunnan Academy of Agricultural Sciences, Kunming 650223, China.

Abstract

The coat protein (CP) is an important structural protein that plays many functional roles during the viral cycle. In this study, the CP of pepper mild mottle virus (PMMoV) was genetically fused to GFP using the foot-and-mouth disease virus peptide 2A linker peptide and the construct (PMMoV-GFP^2A) was shown to be infectious. The systemic spread of the virus was monitored by its fluorescence in infected plants. Electron microscopy and immunocolloidal gold labelling confirmed that PMMoV-GFP^2A forms rod-shaped particles on which GFP is displayed. Studies of tissue ultrastructure and virion self-assembly confirmed that PMMoV-GFP^2A could be used to monitor the real-time dynamic changes of CP location during virus infection. Aggregations of GFP-tagged virions appeared as fluorescent plaques in confocal laser microscopy. Altogether, PMMoV-GFP^2A is a useful tool for studying the spatial and temporal changes of PMMoV CP during viral infection.

Keywords: 2A; coat protein; pepper mild mottle virus; self-assembly.

Abstract

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