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LOCALIZATION OF THE FIRST APPEARANCE OF BARLEY STRIPE MOSAIC VIRUS PROTEIN IN INFECTED WHEAT CELLS (IMMUNOELECTRON MICROSCOPY, COMPLEXES, GOLD-IMMUNOGLOBULIN-G)
Abstract
Gold-labelled IgG (immunogold) has been shown to be useful marker for plant viruses in immunoelectron microscopy (IEM) of thin sections and dip preparations. Direct or indirect immunostaining was effective. In postsectioning staining, immunogold bound specifically to barley stripe mosaic virus (BSMV) in infected cells and could be quantitated. In leaf dip preparations, gold-labelled antibody decoration markedly improved the identification of viruses, particularly spherical ones, including luteoviruses, over simple decoration. The distribution of BSMV protein in wheat root and shoot tips was determined by immunogold-silver staining on sections of Lowicryl- or Araldite-embedded plant tissue at the light microscope level. Infected cells were distributed in a mosaic pattern in or near the meristematic regions. Based on positive localization of BSMV-infected cells in infected root tips, the location of BSMV coat protein was studied on alternative ultrathin sections by IEM. By following sequential cell-to-cell infection from the meristematic point back toward older cells, four cytological stages of early infection could be distinguished. In stage I, proplastids contained peripheral vesicles (= infection initiation markers). BSMV protein was first detected on vesiculated proplastid membranes by immunogold. Sequentially, BSM virions appeared mostly perpendicular to the proplastid membrane at stage II and to the endoplasmic reticulum at stage III. In stage IV, viral cost protein appeared in abundance in euchromatin regions of nuclei while few or no virions were present . By gold-nuclease staining, euchromatin was the main location of nuclear DNA. No excess viral protein could be detected in cell extracts of systemically infected tissue. DsRNA was localized at all four stages in vesicles of vesiculated proplastids when anti-poly(I):poly(C) was used as a primary stain followed by immunogold. It was absent from proplastids of healthy cells. This supports a role for proplastids in BSMV replication.
Subject Area
Plant pathology
Recommended Citation
LIN, NA-SHENG, "LOCALIZATION OF THE FIRST APPEARANCE OF BARLEY STRIPE MOSAIC VIRUS PROTEIN IN INFECTED WHEAT CELLS (IMMUNOELECTRON MICROSCOPY, COMPLEXES, GOLD-IMMUNOGLOBULIN-G)" (1984). ETD collection for University of Nebraska-Lincoln. AAI8428207.
https://digitalcommons.unl.edu/dissertations/AAI8428207