Live-Cell Imaging of Rhabdovirus-Induced Morphological Changes in Plant Nuclear Membranes

Goodin, Michael M.; Yelton, Sharon; Ghosh, Debasish; Mathews, Stephanie; Lesnaw, Judith A.

doi:10.1094/mpmi-18-0703

Cited by 42 publications

(45 citation statements)

References 40 publications

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“…However, we failed to observe nuclear accumulation of the recruited ER membranes in the OFV-infected cells (Fig. 1H, arrow), unlike previous reports for SYNV and PYDV (2,39,40). Such nuclear accumulation of ER membranes is believed to be required for SYNV maturation (budding) from the nuclear side into the perinuclear space (41).…”

Section: Discussioncontrasting

confidence: 86%

“…The morphological changes that OFV induces, such as swelling of the cell nucleus, the appearance of a large viroplasm, and exclusion of chromatin and the nucleolus from the viroplasm (Fig. 1), are similar to those induced by two nucleorhabdoviruses, potato yellow dwarf virus (PYDV) and SYNV (39). However, we failed to observe nuclear accumulation of the recruited ER membranes in the OFV-infected cells (Fig.…”

Section: Discussionmentioning

confidence: 57%

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Orchid Fleck Virus Structural Proteins N and P Form Intranuclear Viroplasm-Like Structures in the Absence of Viral Infection

Kondō

Chiba

Andika

et al. 2013

J Virol

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bOrchid fleck virus (OFV) has a unique two-segmented negative-sense RNA genome that resembles that of plant nucleorhabdoviruses. In infected plant cells, OFV and nucleorhabdoviruses induce an intranuclear electron-lucent viroplasm that is believed to be the site for virus replication. In this study, we investigated the molecular mechanism by which OFV viroplasms are produced in vivo. Among OFV-encoded proteins, the nucleocapsid protein (N) and the putative phosphoprotein (P) were present in nuclear fractions of OFV-infected Nicotiana benthamiana plants. Transient coexpression of N and P, in the absence of virus infection, was shown to be sufficient for formation of an intranuclear viroplasm-like structure in plant cells. When expressed independently as a fluorescent protein fusion product in uninfected plant cells, N protein accumulated throughout the cell, while P protein accumulated in the nucleus. However, the N protein, when coexpressed with P, was recruited to a subnuclear region to induce a large viroplasm-like focus. Deletion and substitution mutagenesis demonstrated that the P protein contains a nuclear localization signal (NLS). Artificial nuclear targeting of the N-protein mutant was insufficient for formation of viroplasm-like structures in the absence of P. A bimolecular fluorescence complementation assay confirmed interactions between the N and P proteins within subnuclear viroplasm-like foci and interactions of two of the N. benthamiana importin-␣ homologues with the P protein but not with the N protein. Taken together, our results suggest that viroplasm formation by OFV requires nuclear accumulation of both the N and P proteins, which is mediated by P-NLS, unlike nucleorhabdovirus viroplasm utilizing the NLS on protein N. R habdoviruses (family Rhabdoviridae, order Mononegavirales) have a broad range of hosts, including humans, livestock, fish, plants, and insects (1). They have bacilliform or bulletshaped virions containing a nonsegmented negative-sense RNA [(Ϫ)RNA] as the genome (2). All of the well-characterized animal rhabdoviruses replicate in the cytoplasm (1). However, the plant rhabdoviruses are divided into two groups, depending on their sites of replication and morphogenesis. Members of the genus Cytorhabdovirus replicate and undergo morphogenesis in the cytoplasm, whereas members of the genus Nucleorhabdovirus multiply in plant cell nuclei and their virions bud through the inner nuclear envelopes (3, 4).Orchid fleck virus (OFV), a Brevipalpus (Acari: Tenuipalpidae) mite-transmitted virus, has a unique two-segmented (Ϫ)RNA genome that resembles that of plant rhabdoviruses (5, 6). OFV is distributed worldwide and infects many kinds of orchids (5,7,8). OFV RNA1 (GenBank accession number AB244417; 6,413 nucleotides [nt]) contains five genes in the order 3=-N-P-3-M-G-5=, in which N encodes the nucleocapsid protein (open reading frame 1 [ORF1]), P encodes the phosphoprotein (ORF2), gene 3 encodes the candidate movement protein (ORF3), M encodes the matrix protein (ORF4), and G encodes the can...

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Section: Discussioncontrasting

confidence: 86%

Section: Discussionmentioning

confidence: 57%

Orchid Fleck Virus Structural Proteins N and P Form Intranuclear Viroplasm-Like Structures in the Absence of Viral Infection

Kondō

Chiba

Andika

et al. 2013

J Virol

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“…In addition to speed, another great advantage of agroinfiltration in N. benthamiana is the ease by which dyes such as 4#,6-diamidino-2-phenylindole (DAPI), to stain DNA Goodin et al, 2007; Fig. 2, A-F), or BODIPY-TR, to stain endomembranes (Goodin et al, 2005), can be infiltrated into leaves prior to examination of tissue samples by microscopy. This approach serves to improve both the quality and accuracy of interpretation of micrographs, thus avoiding the low quality green dots on black images that lack any frame of reference to assist interpretation of the data.…”

Section: New Gateway-compatible Vectors For Expression Of Autofluoresmentioning

confidence: 99%

“…The examples discussed above provide great confidence that agroinfiltration-mediated transient expression of proteins provides data comparable to that obtained using transgenic plants. Therefore, in addition to the essential task of localizing the proteome, transient expression assays in plants will continue to reveal the subtleties of proteins and membrane dynamics in pathogen-infected cells which, ultimately, will aid in establishing the relationship between pathogen-induced changes in protein localization and gene expression (Goodin et al, 2005).…”

Section: Accuracy Of Protein Localization: Transient Expression Compamentioning

confidence: 99%

New Gateways to Discovery

et al. 2007

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“…Apart from measuring disease, it should be emphasized that image analysis has been widely applied to study plant pathogen dispersal, colonization and infection processes at the microscopic level [153][154][155][156][157]. Image analysis of pathogen progress (using Green Fluorescent Protein [GFP] and related biofluorescence markers) combined with epifluorescence microscopy and laser scanning confocal microscopy has been used successfully to study many different host-pathogen systems [158][159][160][161][162][163][164][165][166][167][168].…”

Section: Application Of Visible-wavelength Photography and Image Analmentioning

confidence: 99%

Detection and measurement of plant disease symptoms using visible-wavelength photography and image analysis.

Bock¹,

Nutter²

2011

CABI Reviews

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Plant disease detection and severity assessment are required for many purposes, including predicting yield loss, monitoring and forecasting epidemics, judging host resistance and for studying fundamental biological host-pathogen processes. If assessments of disease severity are inaccurate and/or imprecise, incorrect conclusions might be drawn and incorrect actions taken. Image analysis based on digital images made using visible wavelengths is one of the several methods used to detect and quantify disease; it offers advantages compared with visual assessment or other methods. Over the last 30 years, major advances have been made to improve reliability, precision and accuracy of image analysis for detecting and measuring plant disease. Although the equipment and software continue to become more sophisticated, these technologies are also becoming easier to use. As a result, image analysis to measure plant disease is becoming increasingly widely used, and has now been applied in the study of numerous plant diseases. This review describes the history, technology and application of visible-wavelength photography and image analysis, and progress towards realizing the full potential of these systems in plant disease detection and assessment RightsWorks produced by employees of the U.S. Government as part of their official duties are not copyrighted within the U.S. The content of this document is not copyrighted. AbstractPlant disease detection and severity assessment are required for many purposes, including predicting yield loss, monitoring and forecasting epidemics, judging host resistance and for studying fundamental biological host-pathogen processes. If assessments of disease severity are inaccurate and/or imprecise, incorrect conclusions might be drawn and incorrect actions taken. Image analysis based on digital images made using visible wavelengths is one of the several methods used to detect and quantify disease; it offers advantages compared with visual assessment or other methods. Over the last 30 years, major advances have been made to improve reliability, precision and accuracy of image analysis for detecting and measuring plant disease. Although the equipment and software continue to become more sophisticated, these technologies are also becoming easier to use. As a result, image analysis to measure plant disease is becoming increasingly widely used, and has now been applied in the study of numerous plant diseases. This review describes the history, technology and application of visible-wavelength photography and image analysis, and progress towards realizing the full potential of these systems in plant disease detection and assessment.Keywords: Digital images, Disease incidence, Disease severity, Image analysis, Plant disease assessment, Rater error, Remote sensing, Visible spectrumReview Methodology: We used existing reviews (cited herein) as a source to locate many of the articles. More recent articles were located through online searches using CAB abstracts, Agricola and Web of Science. Keywor...

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Live-Cell Imaging of Rhabdovirus-Induced Morphological Changes in Plant Nuclear Membranes

Cited by 42 publications

References 40 publications

Orchid Fleck Virus Structural Proteins N and P Form Intranuclear Viroplasm-Like Structures in the Absence of Viral Infection

Orchid Fleck Virus Structural Proteins N and P Form Intranuclear Viroplasm-Like Structures in the Absence of Viral Infection

New Gateways to Discovery

Detection and measurement of plant disease symptoms using visible-wavelength photography and image analysis.

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