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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For immunofluorescence microscopy, polyclonal anti-2B antibodies named anti-MP raised in rabbits as described in 53 were used at a 1: In an attempt to visualize the membranes that cosedimented with the VPg precursors, ISEM was performed on sucrose gradient fractions with immunoaffinity-purified anti-VPg antibodies. Yellow mottling may sometimes accompany foliar deformations. Fruit clusters are reduced in size and number with irregular ripening.
VPg Northern-immunoblots as a means for detection of viral RNAs in protoplasts or plants infected with grapevine fanleaf nepovirus. Effect of cerulenin on the synthesis of very-long-chain fatty acids in microsomes from leek seedlings.
Grapevine Fanleaf Degeneration Disease
Panel J is a 2. Panels F, J, and U correspond to healthy cells. Articles with ‘species’ microformats All stub articles. Genome organization of grapevine fanleaf nepovirus RNA2 deduced from the K polyprotein P2 in vitro cleavage products. The grapeivne Golgi apparatus: The procedure was adapted from Bienz et al. These spots were always restricted to the VPg-labeled areas in the nuclear periphery Fig.
In the first syndrome, infectious malformations, the vines may be stunted or show reduced vigor. Protoplast viability in the absence or grappevine of drugs was assessed 48 grapeevine after transfection by using FDA staining.
ISEM of sucrose gradient fractions. One 10 6 protoplasts were harvested 72 h postinfection hpi and homogenized in 1 ml of modified buffer Q 59 containing 50 mM Tris-HCl pH 8. No electron-dense structures similar to those described in CPMV-infected cells 15 were observed. RNA1 encodes polyprotein P1 kDawhich is processed by an embedded proteinase activity into five proteins required for replication, namely, 1A of unknown function1B probably the helicase1C VPg grapevune, 1D proteinaseand 1E polymerase 38 ganleaf, Upon uptake of the virus, a nematode can remain viruliferous for nearly a year.
Evidence for the involvement of virus-induced vesicles in poliovirus replication was obtained already in the sixties 10and the molecular mechanisms fznleaf the formation of virus-induced vesicles, their origin, and their role in viral replication were recently unraveled 1958 To further characterize the GFLV-induced vesicles, crude extracts of healthy and infected T-BY2 protoplasts were fractionated in a linear sucrose gradient.
Small vesicles were specifically immunotrapped by anti-VPg from ER-enriched bottom fractions of the sucrose gradient from infected protoplasts. Intracellular localization of poliovirus plus- and minus-strand RNA visualized by strand-specific fluorescent in situ hybridization. These observations imply that GFLV replication depends both on ER-derived membrane recruitment and on de novo lipid synthesis.
In contrast to proteins involved in viral replication, the 2B movement protein and, to a lesser extent, the 2C coat protein were not confined to the viral compartment but were transported toward the cell periphery, a finding consistent with their role virys cell-to-cell movement of virus particles.
Welcome This is where you can find research-based information from America’s land-grant universities enabled by eXtension. Finally, the CP-containing spikes in Fig.
Cowpea mosaic ivrus infection induces a massive proliferation of endoplasmic reticulum but not Golgi membranes and is dependent on de novo membrane synthesis. In some cases, the transmembrane domains responsible for their anchoring on specific membranes could be identified 18 fanlaef, 3556 allowing the formation of dedicated structures where coupled translation and synthesis of both minus- and plus-strand RNA take place 8. Immunocytochemical localization of TYMV-coded structural and non-structural proteins by the protein A-gold technique.
Vanleaf, Peter D, ed. The sedimentation of Golgi Fig. Observations were done under an epifluorescence microscope Nikon Eclipse E with adequate filters.
Bottom fractions enriched in ER rather than Golgi markers contained also the and kDa VPg precursors and electron microscopy of these fractions revealed the presence of numerous membraneous vesicles.
Grapevine fanleaf virus – Wikipedia
This article has been cited by other articles in PMC. The ER aggregates and anti-VPg labeling colocalized perfectly as shown by the yellow signal obtained upon merging the two pictures Fig. The number of infected cells, as revealed by the anti-VPg or anti-CP signal, was counted versus the total number of protoplasts determined under UV illumination vius staining the nucleus with Hoechst and grapevinee for cell viability.
Sequence analysis and coat protein cistron location”. Such complexes probably ensure protection of the viral RNA being synthesized from degradation by cellular RNases.