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Searching for Phytoferritin in Unstained Plant Sections

Iron is one of the most important transition metals for life on Earth. It is essential for both respiration and photosynthesis. Healthy bacterial, plant and animal cells need a well-controlled concentration of iron, storing the excess in oxidized bio-available form with the use of the iron storage protein ferritin. Our study focuses on phytoferritin and aims to determine the structure of the oxidized iron stored inside plant cells. We recently performed electron diffraction experiments to discover nanocrystalline botanical magnetite (Fe3O4) [1], haematite (Fe2O3), ε-Fe2O3, and 3CaO·Fe2O3·H2O [2] in biochemical extracts from grass cells. In combination with a modified centrifugation procedure, and with TEM imaging, we also found the first evidence for correlation between iron nanocrystal structure and modes of biological self-assembly [2]. To confirm that the biochemical extraction does not affect the identification of botanical iron biominerals, it is important to develop methods that will allow electron diffraction studies of phytoferritin in unstained sectioned plant cells. Here we report our first success in this direction, using an iron starvation followed by re-supply treatment that induces formation of phytoferritin in new growth.