Simple hand-held devices for the efficient infection of plants with viral-encoding constructs by particle bombardment

Gal‐On, Amit; Meiri, Eti; Elman, C.; Gray, D. J.; Gaba, Victor

doi:10.1016/s0166-0934(96)02146-5

Cited by 75 publications

(39 citation statements)

References 23 publications

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“…Cotyledons of 5-day-old plants were inoculated with a plasmid containing an infectious cDNA encoding the full-length ZYMV under control of the 35S Cauliflower mosaic virus promoter (17). Inoculation was done by particle bombardment with a handheld device, the handgun (16). Squash leaves were collected on ice at different time points after inoculation for RNA and protein extraction.…”

Section: Methodsmentioning

confidence: 99%

The Conserved FRNK Box in HC-Pro, a Plant Viral Suppressor of Gene Silencing, Is Required for Small RNA Binding and Mediates Symptom Development

Shiboleth¹,

Haronsky²,

Leibman³

et al. 2007

J Virol

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187

148

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Section: Methodsmentioning

confidence: 99%

The Conserved FRNK Box in HC-Pro, a Plant Viral Suppressor of Gene Silencing, Is Required for Small RNA Binding and Mediates Symptom Development

Shiboleth¹,

Haronsky²,

Leibman³

et al. 2007

J Virol

Self Cite

187

148

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“…Plasmid DNAs and transcripts from viral constructs were introduced into source leaf epidermal cells by particle bombardment using a Handgun essentially as described by Gal-On et al (1997). Briefly, ;2 mg of plasmid DNA or RNA transcript was mixed with 5 mL of ethanol and 0.55 mg of 1-mm gold micro carrier (Bio-Rad Laboratories, Hercules, CA) in ethanol to give a final volume of 18 mL.…”

Section: Microprojectile Bombardmentmentioning

confidence: 99%

Two Plant–Viral Movement Proteins Traffic in the Endocytic Recycling Pathway

Haupt¹,

Cowan²,

Ziegler³

et al. 2005

The Plant Cell

195

157

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Many plant viruses exploit a conserved group of proteins known as the triple gene block (TGB) for cell-to-cell movement. Here, we investigated the interaction of two TGB proteins (TGB2 and TGB3) of Potato mop-top virus (PMTV), with components of the secretory and endocytic pathways when expressed as N-terminal fusions to green fluorescent protein or monomeric red fluorescent protein (mRFP). Our studies revealed that fluorophore-labeled TGB2 and TGB3 showed an early association with the endoplasmic reticulum (ER) and colocalized in motile granules that used the ER-actin network for intracellular movement. Both proteins increased the size exclusion limit of plasmodesmata, and TGB3 accumulated at plasmodesmata in the absence of TGB2. TGB3 contains a putative Tyr-based sorting motif, mutations in which abolished ER localization and plasmodesmatal targeting. Later in the expression cycle, both fusion proteins were incorporated into vesicular structures. TGB2 associated with these structures on its own, but TGB3 could not be incorporated into the vesicles in the absence of TGB2. Moreover, in addition to localization to the ER and motile granules, mRFP-TGB3 was incorporated into vesicles when expressed in PMTV-infected epidermal cells, indicating recruitment by virus-expressed TGB2. The TGB fusion protein-containing vesicles were labeled with FM4-64, a marker for plasma membrane internalization and components of the endocytic pathway. TGB2 also colocalized in vesicles with Ara7, a Rab5 ortholog that marks the early endosome. Protein interaction analysis revealed that recombinant TGB2 interacted with a tobacco protein belonging to the highly conserved RME-8 family of J-domain chaperones, shown to be essential for endocytic trafficking in Caenorhabditis elegans and Drosophila melanogaster. Collectively, the data indicate the involvement of the endocytic pathway in viral intracellular movement, the implications of which are discussed.

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“…The bombardment method was essentially as described by Gal-On et al (1997). Briefly, 5 L of ethanol was added to ‫2ف‬ g of plasmid DNA or RNA transcript.…”

Section: Microprojectile Bombardmentmentioning

confidence: 99%

Functional Analysis of a DNA-Shuffled Movement Protein Reveals That Microtubules Are Dispensable for the Cell-to-Cell Movement of Tobacco mosaic virus

Gillespie¹,

Boevink²,

Haupt³

et al. 2002

Plant Cell

162

171

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Simple hand-held devices for the efficient infection of plants with viral-encoding constructs by particle bombardment

Cited by 75 publications

References 23 publications

The Conserved FRNK Box in HC-Pro, a Plant Viral Suppressor of Gene Silencing, Is Required for Small RNA Binding and Mediates Symptom Development

The Conserved FRNK Box in HC-Pro, a Plant Viral Suppressor of Gene Silencing, Is Required for Small RNA Binding and Mediates Symptom Development

Two Plant–Viral Movement Proteins Traffic in the Endocytic Recycling Pathway

Functional Analysis of a DNA-Shuffled Movement Protein Reveals That Microtubules Are Dispensable for the Cell-to-Cell Movement of Tobacco mosaic virus

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