Award details

Imaging the early events of virus infection in plants

Reference	BB/E001564/1
Principal Investigator / Supervisor	Professor Karl Oparka
Co-Investigators / Co-Supervisors
Institution	University of Edinburgh
Department	Sch of Biological Sciences
Funding type	Research
Value (£)	361,098
Status	Completed
Type	Research Grant
Start date	01/10/2006
End date	30/09/2009
Duration	36 months

Abstract

The aim of this project is to image the earliest events of virus infection using real-time imaging approaches, and to identify the host factors that interact with viral RNA during its intra-and intercellular transport. Tobacco mosaic virus (TMV) viral RNA will be labelled with Alexa-UTP and microinjected into trichome cells to image the earliest events in viral RNA anchoring and trafficking. To image newly translated pools of viral gene products, viral vectors will be constructed that express the movement protein, coat protein and replicase proteins tagged with tetracysteine motifs that will be subsequently localized with FlAsH and ReAsH reagents. The project will also examine the phenomenon by which 'rescue' of viral RNA occurs in the presence of heterologous movement proteins using a combination of microinjection and deletion mutagenesis. The project will utilise real-time deconvolution imaging approaches for RNA tracking developed in the Central Optical Imaging Laboratory (COIL), Edinburgh University.

Summary

This project will investigate the location and redistribution of fluorescent viral RNA immediately following its microinjection into living plant cells. The project will utilise real-time imaging approaches and newly developed FlAsH and ReAsH tags to track viral gene products immediately following their translation. These tagged proteins will be imaged against a transgenic background in which a range of cellular components (e.g. ER, microtubules, plasmodesmata) have been stably highlighted with green fluorescent protein (GFP). The aim is to provide unequivocal evidence for the pathway of viral RNA trafficking in plants during the first stages of infection, and to identify the viral and host factors that interact with the viral genome during transit. The project will be carried out in collaboration with Dr Ilan Davis, Wellcome Centre for Cell Biology, and will utilise real-time deconvolution imaging approaches developed in the Central Optical Imaging Laboratory (COIL), Edinburgh University.

Committee	Closed Committee - Plant & Microbial Sciences (PMS)
Research Topics	Crop Science, Microbiology, Plant Science
Research Priority	X – Research Priority information not available
Research Initiative	X - not in an Initiative
Funding Scheme	X – not Funded via a specific Funding Scheme

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