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Research article2017Peer reviewedOpen access

Salicylic acid interferes with GFP fluorescence in vivo

de Jonge, Jennifer; Hofius, Daniel; Hennig, Lars

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; fluorescence microscopy; fluorescent protein; GFP fusion proteins; histones; salicylic acid

Published in

Journal of Experimental Botany
2017, Volume: 68, number: 7, pages: 1689-1696 Publisher: OXFORD UNIV PRESS

      SLU Authors

      • Associated SLU-program

        SLU Plant Protection Network

        UKÄ Subject classification

        Botany

        Publication identifier

        DOI: https://doi.org/10.1093/jxb/erx031

        Permanent link to this page (URI)

        https://res.slu.se/id/publ/82821