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A genetic system for Citrus Tristeza Virus using the non-natural host Nicotiana benthamiana: an update

Overview of attention for article published in Frontiers in Microbiology, January 2013
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Title
A genetic system for Citrus Tristeza Virus using the non-natural host Nicotiana benthamiana: an update
Published in
Frontiers in Microbiology, January 2013
DOI 10.3389/fmicb.2013.00165
Pubmed ID
Authors

Silvia Ambrós, Susana Ruiz-Ruiz, Leandro Peña, Pedro Moreno

Abstract

In nature Citrus tristeza virus (CTV), genus Closterovirus, infects only the phloem cells of species of Citrus and related genera. Finding that the CTV T36 strain replicated in Nicotiana benthamiana (NB) protoplasts and produced normal virions allowed development of the first genetic system based on protoplast transfection with RNA transcribed from a full-genome cDNA clone, a laborious and uncertain system requiring several months for each experiment. We developed a more efficient system based on agroinfiltration of NB leaves with CTV-T36-based binary plasmids, which caused systemic infection in this non-natural host within a few weeks yielding in the upper leaves enough CTV virions to readily infect citrus by slash inoculation. Stem agroinoculation of citrus and NB plants with oncogenic strains of Agrobacterium tumefaciens carrying a CTV-T36 binary vector with a GUS marker, induced GUS positive galls in both species. However, while most NB tumors were CTV positive and many plants became systemically infected, no coat protein or viral RNA was detected in citrus tumors, even though CTV cDNA was readily detected by PCR in the same galls. This finding suggests (1) strong silencing or CTV RNA processing in transformed cells impairing infection progress, and (2) the need for using NB as an intermediate host in the genetic system. To maintain CTV-T36 in NB or assay other CTV genotypes in this host, we also tried to graft-transmit the virus from infected to healthy NB, or to mechanically inoculate NB leaves with virion extracts. While these trials were mostly unsuccessful on non-treated NB plants, agroinfiltration with silencing suppressors enabled for the first time infecting NB plants by side-grafting and by mechanical inoculation with virions, indicating that previous failure to infect NB was likely due to virus silencing in early infection steps. Using NB as a CTV host provides new possibilities to study virus-host interactions with a simple and reliable system.

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Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 21 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 21 100%

Demographic breakdown

Readers by professional status Count As %
Student > Ph. D. Student 5 24%
Researcher 3 14%
Professor 3 14%
Student > Master 2 10%
Student > Bachelor 1 5%
Other 3 14%
Unknown 4 19%
Readers by discipline Count As %
Agricultural and Biological Sciences 10 48%
Biochemistry, Genetics and Molecular Biology 6 29%
Pharmacology, Toxicology and Pharmaceutical Science 1 5%
Neuroscience 1 5%
Unknown 3 14%
Attention Score in Context

Attention Score in Context

This research output has an Altmetric Attention Score of 1. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 02 July 2013.
All research outputs
#20,195,877
of 22,713,403 outputs
Outputs from Frontiers in Microbiology
#22,136
of 24,542 outputs
Outputs of similar age
#248,765
of 280,747 outputs
Outputs of similar age from Frontiers in Microbiology
#264
of 407 outputs
Altmetric has tracked 22,713,403 research outputs across all sources so far. This one is in the 1st percentile – i.e., 1% of other outputs scored the same or lower than it.
So far Altmetric has tracked 24,542 research outputs from this source. They typically receive a little more attention than average, with a mean Attention Score of 6.4. This one is in the 1st percentile – i.e., 1% of its peers scored the same or lower than it.
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We're also able to compare this research output to 407 others from the same source and published within six weeks on either side of this one. This one is in the 1st percentile – i.e., 1% of its contemporaries scored the same or lower than it.