Infection by Grapevine fanleaf nepovirus (GFLV), a bipartite RNA virus of positive polarity belonging to the Comoviridae family, causes extensive cytopathic. The specific transmission of Grapevine fanleaf virus by its nematode vector Xiphinema index is solely determined by the viral coat protein. There are plenty of plant viruses that no one has heard of, but few are as widely known as grapevine fanleaf virus. Learn how to identify a sick.

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Brome mosaic virus RNA replication proteins 1a and 2a colocalize and 1a independently localizes on the yeast endoplasmic reticulum. The external surface of the vesicles was systematically coated with electron-dense material Fig.

In contrast to uninfected cells, in which plastids and mitochondria were found surrounding the nucleus Fig. Nepoviruses Viral grape diseases Viral plant disease stubs.

Among these species, only the and kDa proteins clearly sedimented into the gradient Fig. No signal was detected in total grqpevine extracts from healthy protoplasts Fig.

For immunofluorescence microscopy, polyclonal anti-2B antibodies named anti-MP raised in rabbits as described in 53 were used at a 1: PDB entry 2y26 [1].

To correct for variations in protoplast viability and percentage of infection between independent experiments, results were expressed as the ratio between the percentage of infection and the percentage of viable cells. Differential interference contrast observations were superimposed in panels C, D, F, and M for visualization of the cell and its nucleus.


Grapevine Fanleaf Virus Replication Occurs on Endoplasmic Reticulum-Derived Membranes

Photo courtesy of William M. This article has been cited by other articles in PMC.

Labeling and detection of nascent RNA. Further in vivo virsu course analysis will be required to confirm the observed variations in the number of Golgi stacks within a single cell during the course of infection, but this is beyond the scope of this study. To further characterize the GFLV-induced vesicles, crude extracts of healthy and infected T-BY2 protoplasts were fractionated in a linear sucrose gradient.

Tubular structures involved in movement of cowpea mosaic virus are also formed in infected cowpea protoplasts.

Using epifluorescence microscopy, we have previously described the formation of a perinuclear complex where viral RNA was synthesized and viral proteins accumulated 23but this could not be further analyzed due to technical limitations. Additional ultrastructural alterations accompanied the modifications of the endomembrane system.

Grapevine Fanleaf Virus Replication Occurs on Endoplasmic Reticulum-Derived Membranes

The sedimentation of Golgi Fig. The longer polyprotein the product of RNA1 is cleaved into five functional proteins, including the viral RNA polymerase [4] and the shorter from RNA2 is cleaved into three proteins. Characterization of the poliovirus replication complex. The type of fanlesf involved in replication depends on the virus considered: National Center for Biotechnology InformationU. Intracellular localization of poliovirus plus- and minus-strand RNA visualized by strand-specific fluorescent in situ hybridization.


Such complexes probably ensure protection of the viral RNA being synthesized from degradation by cellular RNases. The second syndrome, yellow mosaic, is caused by chromogenic virus strains. These observations imply that GFLV replication depends both on Grapevinf membrane recruitment and on de novo lipid synthesis.

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Moreover, the use of transgenic T-BY2 cell lines expressing green fluorescent protein GFP targeted to the endoplasmic reticulum ER and the Golgi apparatus GA provided a direct means to follow the virus-induced modifications of the endomembrane system and allowed us to conclude that GFLV replicates in association with specific membranes mainly derived from ER. Journal List J Virol v.

Sequence analysis and coat protein cistron location”. Please review our privacy policy. Cerulenin, a drug inhibiting de novo synthesis of phospholipids, also inhibited GFLV replication.

View publishing information about this page. Preparation and infection of protoplasts. Author information Article notes Copyright and License information Disclaimer. The cell lines used were: We hypothesized that the 2A grapevinw of polyprotein P2 could mediate the transport of the nascent P2-RNA2 complexes from their initial location in the cytosol to the perinuclear replication sites where RNA2 replication and P2 cleavage take place