Salicylic acid interferes with GFP fluorescence in vivo
- PMID: 28369601
- PMCID: PMC5441896
- DOI: 10.1093/jxb/erx031
Salicylic acid interferes with GFP fluorescence in vivo
Abstract
Fluorescent proteins have become essential tools for cell biologists. They are routinely used by plant biologists for protein and promoter fusions to infer protein localization, tissue-specific expression and protein abundance. When studying the effects of biotic stress on chromatin, we unexpectedly observed a decrease in GFP signal intensity upon salicylic acid (SA) treatment in Arabidopsis lines expressing histone H1-GFP fusions. This GFP signal decrease was dependent on SA concentration. The effect was not specific to the linker histone H1-GFP fusion but was also observed for the nucleosomal histone H2A-GFP fusion. This result prompted us to investigate a collection of fusion proteins, which included different promoters, subcellular localizations and fluorophores. In all cases, fluorescence signals declined strongly or disappeared after SA application. No changes were detected in GFP-fusion protein abundance when fluorescence signals were lost indicating that SA does not interfere with protein stability but GFP fluorescence. In vitro experiments showed that SA caused GFP fluorescence reduction only in vivo but not in vitro, suggesting that SA requires cellular components to cause fluorescence reduction. Together, we conclude that SA can interfere with the fluorescence of various GFP-derived reporter constructs in vivo. Assays that measure relocation or turnover of GFP-tagged proteins upon SA treatment should therefore be evaluated with caution.
Keywords: Arabidopsis; GFP fusion proteins; fluorescence microscopy; fluorescent protein; histones; salicylic acid..
© The Author 2017. Published by Oxford University Press on behalf of the Society for Experimental Biology.
Figures
References
-
- Ascenzi R, Gantt JS. 1997. A drought-stress-inducible histone gene in Arabidopsis thaliana is a member of a distinct class of plant linker histone variants. Plant Molecular Biology 34, 629–641. - PubMed
-
- Boursiac Y, Boudet J, Postaire O, Luu DT, Tournaire-Roux C, Maurel C. 2008. Stimulus-induced downregulation of root water transport involves reactive oxygen species-activated cell signalling and plasma membrane intrinsic protein internalization. The Plant Journal 56, 207–218. - PubMed
-
- Brunoud G, Wells DM, Oliva M, et al. 2012. A novel sensor to map auxin response and distribution at high spatio-temporal resolution. Nature 482, 103–106. - PubMed
-
- Chalfie M, Tu Y, Euskirchen G, Ward WW, Prasher DC. 1994. Green fluorescent protein as a marker for gene expression. Science 263, 802–805. - PubMed
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
