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Published on: May 14, 2008
Aphid Acquisition and Cellular Transport of Potato leafroll virus-like Particles Lacking P5 Readthrough Protein
Phytopathology
|October 24, 2008
Summary
Potato leafroll virus (PLRV) coat protein can form virus-like particles (VLPs) without the P5 protein. These PLRV VLPs are transported through aphid vectors, suggesting P5 is not essential for insect transmission.
Area of Science:
- Plant Virology
- Insect Vector Biology
- Molecular Virology
Background:
- Potato leafroll virus (PLRV) is a significant plant pathogen transmitted by aphids.
- The PLRV particle comprises coat protein (CP) and a P5 readthrough protein.
- The role of the P5 protein in aphid transmission is not fully understood.
Purpose of the Study:
- To investigate the necessity of the PLRV P5 readthrough protein for virus-like particle (VLP) formation and aphid transmission.
- To characterize VLPs produced in insect cells using recombinant baculoviruses.
- To compare the transmission efficiency of PLRV and PLRV VLPs in aphids.
Main Methods:
- Generation of recombinant baculoviruses expressing PLRV CP (with His-tag) or PLRV genomic RNA.
- Infection of Spodoptera frugiperda cells to produce VLPs.
- Purification and characterization of VLPs using electron microscopy.
- Aphid (Myzus persicae) feeding and injection experiments with purified PLRV and VLPs.
- Electron microscopic analysis of aphid tissues to track particle transport.
Main Results:
- Recombinant baculovirus expressing PLRV CP produced His-tagged VLPs (P3-6H).
- Recombinant baculovirus expressing PLRV RNA did not produce VLPs.
- Co-infection resulted in VLPs containing PLRV RNA packaged in P3-6H CP (FL particles).
- Both P3-6H and FL particles were morphologically similar to wild-type PLRV but lacked P5 protein.
- Aphids exposed to purified PLRV or VLPs showed no differences in particle uptake, transport, or exocytosis in midgut and salivary glands.
- Particles were observed in aphid salivary canals, indicating potential for transmission.
Conclusions:
- PLRV coat protein can assemble into infectious-like particles independently of the P5 readthrough protein.
- The P5 readthrough protein may not be essential for the cellular transport of PLRV within the aphid vector.
- These findings provide insights into the structural requirements for PLRV transmission by aphids.
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