Visualization of plant viral suppressor silencing activity in intact leaf lamina by quantitative fluorescent imaging

Simple item page
dc.contributor.authorStephan, Dirk
dc.contributor.authorSlabber, Coba
dc.contributor.authorGeorge, Gavin
dc.contributor.authorNinov, Victor
dc.contributor.authorFrancis, Kevin P.
dc.contributor.authorBurger, Johan T.
dc.date.accessioned2011-09-06T12:42:43Z
dc.date.available2011-09-06T12:42:43Z
dc.date.issued2011-08
dc.date.updated2011-09-01T05:26:50Z
dc.descriptionPublication of this article was funded by the Stellenbosch University Open Access Fund.en_ZA
dc.descriptionThe original publication is available at http://www.plantmethods.com/content/7/1/25
dc.description.abstractABSTRACT: BACKGROUND: Transient expression of proteins in plants has become a favoured method over the production of stably transformed plants because, in addition to enabling high protein yields, it is both fast and easy to apply. An enhancement of transient protein expression can be achieved by plant virus-encoded RNA silencing suppressor proteins. Since viral suppressor proteins differ in their efficiency to enhance transient protein expression in plants, we developed a whole-leaf green fluorescent protein (GFP)-based imaging assay to quantitatively assess suppressor protein activity. RESULTS: In a transient GFP-expression assay using wild-type and GFP-transgenic N. benthamiana, addition of the plant viral suppressors Beet mild yellowing virus (BMYV-IPP) P0 or Plum pox virus (PPV) HC-Pro was shown to increase fluorescent protein expression 3-4-fold, 7 days post inoculation (dpi) when compared to control plants. In contrast, in agroinfiltrated patches without suppressor activity, near complete silencing of the GFP transgene was observed in the transgenic N. benthamiana at 21 dpi. Both co-infiltrated suppressors significantly enhanced GFP expression over time, with HC-Pro co-infiltrations leading to higher short term GFP fluorescence (at 7 dpi) and P0 giving higher long term GFP fluorescence (at 21 dpi). Additionally, in contrast to HC-Pro co-infiltrations, an area of complete GFP silencing was observed at the edge of P0 co-infiltrated areas. CONCLUSIONS: Fluorescence imaging of whole intact leaves proved to be an easy and effective method for spatially and quantitatively observing viral suppressor efficiency in plants. This suppressor assay demonstrates that plant viral suppressors greatly enhanced transient GFP expression, with P0 showing a more prolonged suppressor activity over time than HC-Pro. Both suppressors could prove to be ideal candidates for enhancing target protein expression in plants.en_ZA
dc.description.versionPublishers' versionen_ZA
dc.format.extent9 p. : ill.
dc.identifier.citationStephan, D., et al. 2011, Visualization of plant viral suppressor silencing activity in intact leaf lamina by quantitative fluorescent imaging, Plant Methods, 7(1) : 25, http://dx.doi.org/10.1186/1746-4811-7-25en_ZA
dc.identifier.issn1746-4811
dc.identifier.otherhttp://dx.doi.org/10.1186/1746-4811-7-25
dc.identifier.urihttp://hdl.handle.net/10019.1/16350
dc.language.isoen_ZAen_ZA
dc.language.rfc3066en
dc.publisherBioMed Central Ltd.en_ZA
dc.rights.holderBioMed Central Ltd.en_ZA
dc.subjectPlant proteinsen_ZA
dc.subjectRNA silencingen_ZA
dc.subjectTransient protein expressionen_ZA
dc.subjectFluorescence imagingen_ZA
dc.titleVisualization of plant viral suppressor silencing activity in intact leaf lamina by quantitative fluorescent imagingen_ZA
dc.typeArticleen_ZA
Files
Original bundle
Now showing 1 - 2 of 2
Thumbnail Image
Name:
1746-4811-7-25.xml
Size:
60.97 KB
Format:
Extensible Markup Language
Description:
Download
Thumbnail Image
Name:
1746-4811-7-25.pdf
Size:
5.85 MB
Format:
Adobe Portable Document Format
Description:
Download
License bundle
Now showing 1 - 1 of 1
Thumbnail Image
Name:
license.txt
Size:
1.71 KB
Format:
Item-specific license agreed upon to submission
Description:
Download
Collections
Research Articles (Genetics)