Tracking the green invaders: advances in imaging virus infection in plants

Tilsner, Jens; Oparka, Karl J.

doi:10.1042/bj20100372

Cited by 20 publications

(11 citation statements)

References 187 publications

(340 reference statements)

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“…More recently a MYB-related transcription factor, Rosea 1, which is also smaller than GFP, has been placed in TMV as a marker for virus location (Bedoya et al, 2012). Advanced microscopes with super high resolution will allow us to more easily determine whether proteins are interacting or simply co-localizing (Tilsner and Oparka, 2010). Access to the N. benthamiana genome (Bombarely et al, 2012) will allow better identification of gene family members projected to influence virus transport and the ability to target individual members for knockdown, over-expression and labeling.…”

Section: Resultsmentioning

confidence: 99%

The cell biology of Tobacco mosaic virus replication and movement

Liu¹,

Nelson²

2013

Front. Plant Sci.

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Successful systemic infection of a plant by Tobacco mosaic virus (TMV) requires three processes that repeat over time: initial establishment and accumulation in invaded cells, intercellular movement, and systemic transport. Accumulation and intercellular movement of TMV necessarily involves intracellular transport by complexes containing virus and host proteins and virus RNA during a dynamic process that can be visualized. Multiple membranes appear to assist TMV accumulation, while membranes, microfilaments and microtubules appear to assist TMV movement. Here we review cell biological studies that describe TMV-membrane, -cytoskeleton, and -other host protein interactions which influence virus accumulation and movement in leaves and callus tissue. The importance of understanding the developmental phase of the infection in relationship to the observed virus-membrane or -host protein interaction is emphasized. Utilizing the latest observations of TMV-membrane and -host protein interactions within our evolving understanding of the infection ontogeny, a model for TMV accumulation and intracellular spread in a cell biological context is provided.

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Section: Resultsmentioning

confidence: 99%

The cell biology of Tobacco mosaic virus replication and movement

Liu¹,

Nelson²

2013

Front. Plant Sci.

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“…Research in plant virology has benefited tremendously from the use of genetically encoded reporter genes (Tilsner and Oparka, 2010). Some markers, when inserted into the viral genome, report on virus infection and movement through the plant.…”

mentioning

confidence: 99%

Visual Tracking of Plant Virus Infection and Movement Using a Reporter MYB Transcription Factor That Activates Anthocyanin Biosynthesis

et al. 2012

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Insertion of reporter genes into plant virus genomes is a common experimental strategy to research many aspects of the viral infection dynamics. Their numerous advantages make fluorescent proteins the markers of choice in most studies. However, the use of fluorescent proteins still has some limitations, such as the need of specialized material and facilities to detect the fluorescence. Here, we demonstrate a visual reporter marker system to track virus infection and movement through the plant. The reporter system is based on expression of Antirrhinum majus MYB-related Rosea1 (Ros1) transcription factor (220 amino acids; 25.7 kD) that activates a series of biosynthetic genes leading to accumulation of colored anthocyanins. Using two different tobacco etch potyvirus recombinant clones tagged with Ros1, we show that infected tobacco (Nicotiana tabacum) tissues turn bright red, demonstrating that in this context, the sole expression of Ros1 is sufficient to induce pigment accumulation to a level readily detectable to the naked eye. This marker system also reports viral load qualitatively and quantitatively by means of a very simple extraction process. The Ros1 marker remained stable within the potyvirus genome through successive infectious passages from plant to plant. The main limitation of this marker system is that color output will depend on each particular plant host-virus combination and must be previously tested. However, our experiments demonstrate accurate tracking of turnip mosaic potyvirus infecting Arabidopsis (Arabidopsis thaliana) and either tobacco mosaic virus or potato X virus infecting Nicotiana benthamiana, stressing the general applicability of the method.

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“…In this approach, the same biological sample is viewed using a number of approaches sequentially from confocal microscopy, through super-resolution microscopy to transmission electron microscopy, bridging important gaps in image scaling between different instruments. The potential for CLEM with respect to imaging plant cells was reviewed recently by Tilsner and Oparka (2010) and Bell and Oparka (2011).…”

Section: Super-resolution Microscopymentioning

confidence: 99%

The structure of the phloem – still more questions than answers

Knoblauch

Oparka

2012

The Plant Journal

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SUMMARYLong-distance assimilate distribution in higher plants takes place in the enucleate sieve-tube system of the phloem. It is generally accepted that flow of assimilates is driven by an osmotically generated pressure differential, as proposed by Ernst Mü nch more than 80 years ago. In the period between 1960 and 1980, the pressure flow hypothesis was challenged when electron microscopic images suggested that sieve tubes contain obstructions that would prevent passive flow. This led to the proposal of alternative translocation mechanisms. However, most investigators came to the conclusion that obstructions in the sieve-tube path were due to preparation artifacts. New developments in bioimaging have vastly enhanced our ability to study the phloem. Unexpectedly, in vivo studies challenge the pressure-flow hypothesis once again. In this review we summarize current investigations of phloem structure and function and discuss their impact on our understanding of long-distance transport in the phloem.

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Tracking the green invaders: advances in imaging virus infection in plants

Cited by 20 publications

References 187 publications

The cell biology of Tobacco mosaic virus replication and movement

The cell biology of Tobacco mosaic virus replication and movement

Visual Tracking of Plant Virus Infection and Movement Using a Reporter MYB Transcription Factor That Activates Anthocyanin Biosynthesis

The structure of the phloem – still more questions than answers

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