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New method for visualization of silica phytoliths in Sorghum bicolor roots by fluorescence microscopy revealed silicate concentration-dependent phytolith formation

Soukup, Milan, Martinka, Michal, Cigáň, Marek, Ravaszová, Frederika, Lux, Alexander
Planta 2014 v.240 no.6 pp. 1365-1372
Sorghum bicolor, endodermis, fluorescence microscopy, image analysis, light microscopy, new methods, pH, phytoliths, roots, silica, sodium silicate
Silica phytoliths are microscopic structures of amorphous hydrated silica (SiO₂·nH₂O) formed by specialized plant cells. Besides their biological roles, physical, chemical, and structural properties of biogenic silica offer a wide spectrum of applications in many fields of industry and technology. Therefore, processes involved in their formation recently become a very interesting topic to study. However, optical transparency and microscopic sizes of silica phytoliths do not allow their visualization and localization by classical light microscopy methods. Their observation thus requires phytolith isolation, technically difficult or lengthy sample preparation procedures, or a work with toxic chemicals. In this paper we are proposing a novel method for visualization of silica phytoliths in Sorghum bicolor root endodermal cells by fluorescence microscopy using alkali mounting solution (pH 12). This method offers an easy and quick preparation of the samples and high contrast imaging. Based on our results we can assume that the proposed fluorescent method for silica phytolith investigation allows observation of multiple samples in relatively short time period and thus might be applicable also for high-throughput screenings. Using this method we found out that after a 3-day cultivation of sorghum plants the minimal needed concentration of sodium silicate, limiting the formation of silica phytoliths in the root endodermis, was 25 µmol dm⁻³. The positive correlation of sodium silicate concentration in the substrate with the phytolith diameter was also observed.